Abstract

Fimbristylis miliacea L. (Family: Cyperaceae) is a plant found in Australia,

Bangladesh, Bhutan, Cambodia, India, Indonesia, Malaysia, Myanmar, Nepal,

Pakistan, Philippines, Sri Lanka, Thailand and Vietnam. No published results are

available in the literature reporting pharmacological activities of this plant. The

present study was aimed to investigate hypoglycemic and hepatoprotective effects

of methanol extract of Fimbristylis miliacea whole plant. Hypoglycemic activity of

the plant extract was examined by oral glucose tolerance test. Hepatoprotective

effect was evaluated by paracetamol induced liver toxicity and liver function

markers (ALT, AST, ALP) and total bilirubin were estimated. The methanolic

extract of Fimbristylis miliacea showed significant hypoglycemic effect. Standard

drug glibenclamide reduced fasting blood glucose concentration by 46.69% after 3 h

while extract at 400 mg/kg showed 36.92% reduction after same time span.

Moderate hepatoprotective effect was obtained against liver damage induced by

paracetamol overdose as evident from decreased serum levels of alanine

aminotransferase, aspartate aminotransferase, alkaline phosphatase and bilirubin in

the Fimbristylis miliacea treated groups (200, 400 and 600 mg/kg) compared to the

intoxicated controls. The standard treatment, silymarin and extract 600 mg/ml

showed as significant decrease in ALT (6.78±.77, p>0.01 & 12.59±5.29, p>0.01

respectively) and ALP (5.12±1.02, p>0.01 & 8.55±1.39) levels compared to

paracetamol control. The results obtained in this study suggest that the methanolic

extract of Fimbristylis miliacea can further be explored for new hypoglycemic and

hepatoprotective agents.

i
 Table of Contents
Abstract .................................................................................................... i
List of Figures ........................................................................................iv
List of Tables .........................................................................................iv
Chapter 1: Introduction ...................................................................... 1
1.1 History of ancient times .............................................................................................. 1
1.2 History of early modern .............................................................................................. 2
1.3 History of middle ages ................................................................................................ 3
1.4 Why are medicinal plants important? ......................................................................... 3
1.5 Exploration of Medicinal Properties of Plants ............................................................ 5
1.6 Contribution of medicinal plants in modern medicine ................................................ 5
1.7 Phytotherapy- development of new synthetic drug ................................................... 11
1.8 Medicinal Plants of Bangladesh ................................................................................ 12
1.8.1 Medicinal plants, its cultivation and Bangladeshi market .................. 13
1.9 Medicinal Plants: Wealth of a Country ..................................................................... 15
1.10 Plant Kingdom: Storehouse of Innumerable Drugs .................................................. 16
1.11 Modern Prescription Drugs ....................................................................................... 17
1.11.1 Herbal Medicine Today ...................................................................... 17
1.11.2 Herbal Drug Research: Bioactivity Guided Approach ....................... 18
1.12 Research in Medicinal Plant...................................................................................... 19
1.12.1 Rationale for Herbal Drug Research in Bangladesh ........................... 21
1.13 Some plants with their traditional uses ..................................................................... 21
1.14 Drug Preparation Based on Medicinal Plant Study................................................... 25
1.15 Drug Development from Plant Source ...................................................................... 26
Chapter 2: Literature review ........................................................... 28
2.1 Introduction to the Plant Family: Cyperaceae ........................................................... 28
2.1.1 Morphology ........................................................................................ 28
2.1.2 Habitat ................................................................................................ 29
2.1.3 Genera of the Cyperaceae Family ....................................................... 30
2.1.4 The plant genus Fribristylis ................................................................ 31
2.1.5 Species: Members of the genus Fimbristylis....................................... 31
2.2 Investigating Plant: Fimbristylis miliacea ................................................................ 35
2.2.1.1 Plant Information ............................................................................ 35

ii
 2.2.2 Taxonomy of Fimbristylis miliacea .................................................... 36
2.2.3 Plant type ............................................................................................ 37
2.2.4 Morphology ........................................................................................ 37
2.2.5 Distribution ......................................................................................... 37
2.2.6 Habitat ................................................................................................. 37
2.2.7 Traditional use of Fimbristylis ............................................................ 37
2.2.8 Literature survey of Fimbristylis miliacea .......................................... 38

Chapter 3: Preparation of plant extract ......................................... 39

3.1 Methods for Extract Preparation ............................................................................... 39
3.1.1 Collection and Identification .............................................................. 39
3.1.2 Preparation of Plant sample ................................................................ 40
3.1.3 Extraction ............................................................................................ 40
3.1.3.1 Cold Extraction Procedure .............................................................. 40
3.1.3.2 Hot Extraction Procedure ................................................................ 40
3.1.3.3 Cold extraction of Fimbristylis miliacea ......................................... 41

Chapter 4: Materials & Methods ..................................................... 42

4.1 Chemicals and drugs ................................................................................................. 42
4.2 Instruments and apparatus ......................................................................................... 42
4.3 Experimental Animals............................................................................................... 42
4.4 Oral glucose tolerance test ........................................................................................ 43
4.5 Hepatoprotective activity .......................................................................................... 43
4.5.1 Experimental design and animal grouping ......................................... 43
4.5.2 Preparation of serum from blood ........................................................ 44
4.5.3 Determination of key liver function biochemical markers ................. 44
4.6 Statistical analysis ..................................................................................................... 45
Chapter 5: Results and discussions.................................................. 46
5.1 Hypoglycemic effect ................................................................................................. 46
5.2 Hepatoprotective effect ............................................................................................. 47
Chapter 6: Conclusion ...................................................................... 51
Chapter 7: Reference ........................................................................ 52

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 List of Figures
Figure 1.1: The Ebers Papyrus (c. 1550 BC) from Ancient Egypt describing the use
of Cannabis sativa applied topically for inflammation ........................... 2
Figure 1.2: Major medicinal plants production zones in Bangladesh....................... 13
Figure 3.1: Fimbristylis miliacea .............................................................................. 39
Figure 5.1: Effect of methanol leaf extract of Fimbristylis miliacea on
hyperglycemic condition. ‘FM’ stands for Fimbristylis miliacea and
100, 200 and 300 indicate doses in mg/kg. A probability level of 0.05 or
less was accepted as significant; *p < 0.05, **
p < 0.01, ***
p < 0.001 vs.
control i.e. distilled water....................................................................... 46

List of Tables
Table 1.1: Plant derived medicinal substance occurring used in modern medicine ... 5
Table 1.2: Some medicinal plants with their traditional uses ................................... 21
Table 5.1: Percentage reduction of blood glucose concentration by methanolic
extract of leaf of Fimbristylis miliacea .................................................. 47
Table 5.2: Effect of Fimbristylis miliacea extract in paracetamol induced liver
damage ................................................................................................... 49

iv
 Chapter 1: Introduction
Every disease that affects mankind has its treatment of a cure occurring naturally on

the earth (Shafikur et all, 2011). From remote antiquity, different parts of plants and

organs of animals have been used as remedies in many civilizations. Medicine is an

ancient art rather than modern science and for that ʺback to natureʺ is a slogan of

recent years.

The World Health Organization (WHO) estimated that 80% of the population of

developing countries relies on traditional medicines, mostly plant drugs for the

primary health care needs. In addition, modern pharmacopoeia still contains at least

25% drugs derived from plants and many others, which are synthetic analogues,

built on prototype. Ever since ancient times, in search for rescue for their disease,

the people looked for drugs in nature. The beginnings of the medicinal plants’ usage

were instinctive, as is the case with animals. In view of the fact that at the time there

was not sufficient information either concerning the reasons for the illnesses or

concerning which plant and how it could be utilized as a cure, everything was based

on experience. In time, the reasons for the usage of specific medicinal plants for

treatment of certain diseases were being discovered; thus, the medicinal plants’

usage gradually abandoned the empiric framework and became founded on

explicatory facts (Petrovska, 2012).

1.1 History of ancient times

The Ebers Papyrus (c. 1550 BC) from Ancient Egypt describes the use of Cannabis

sativa applied topically for inflammation (Figure 1.1). In ancient Sumeria, hundreds

of medicinal plants including myrrh and opium are listed on clay tablets. The

ancient Egyptian Ebers Papyrus lists over 800 plant medicines such as aloe,

1
cannabis, castor bean, garlic, juniper, and mandrake. From ancient times to the

present, Ayurvedic medicine as documented in the Atharva Veda, the Rig Veda and

the Sushruta Samhita has used hundreds of pharmacologically active herbs and

spices such as turmeric, which contains curcumin.

Figure 1.1: The Ebers Papyrus (c. 1550 BC) from Ancient Egypt describing the use
of Cannabis sativa applied topically for inflammation

The Chinese pharmacopoeia, the Shennong Ben Cao Jing records plant medicines

such as chaulmoogra for leprosy, ephedra, and hemp. This was expanded in the

Tang Dynasty Yaoxing Lun. In the fourth century BC, Aristotle's pupil

Theophrastus wrote the first systematic botany text, Historia plantarum. In the first

century AD, the Greek physician Pedanius Dioscorides documented over 1000

recipes for medicines using over 600 medicinal plants in De materia medica; it

remained the authoritative reference on herbalism for over 1500 years, into the

seventeenth century (Collins, 2000).

1.2 History of early modern

The early modern period saw the flourishing of illustrated herbals across Europe,

starting with the 1526 Grete Herball. John Gerard wrote his famous ‘The Herball or

General History of Plants’ in 1597, based on Rembert Dodoens, and Nicholas

2
Culpeper published his The English Physician Enlarged. Many new plant medicines

arrived in Europe as products of Early Modern exploration and the resulting

Columbian Exchange. In Mexico, the sixteenth century Badianus Manuscript

described medicinal plants available in Central America (Gimmel, 2008).

1.3 History of middle ages

In the Early Middle Ages, Benedictine monasteries preserved medical knowledge in

Europe, translating and copying classical texts and maintaining herb gardens.

Hildegard of Bingen wrote Causae et Curae ("Causes and Cures") on medicine. In

the Islamic Golden Age, scholars translated many classical Greek texts including

Dioscorides into Arabic. Herbalism flourished in Baghdad and in Al-Andalus.

Abulcasis (936–1013) of Cordoba wrote The Book of Simples, and Ibn al-Baitar

(1197–1248) recorded hundreds of medicinal herbs such as Aconitum, nux vomica,

and tamarind in his Corpus of Simples. Avicenna included many plants in his 1025

The Canon of Medicine. Abu-Rayhan Biruni, Ibn Zuhr, Peter of Spain, and John of

St Amand wrote further pharmacopoeias (Brater & Daly, 2000).

1.4 Why are medicinal plants important?

A considerable number of definitions have been proposed for medicinal plants.

According to the WHO, A medicinal plant is any plant which, in one or more of its

organs, contains substances that can be used for therapeutic purposes, or which are

precursors for chemo-pharmaceutical semi- synthesis. When a plant is designated as

medicinal, it is implied that the said plant is useful as a drug or therapeutic agent or

active ingredient of a medicinal preparation. Medicinal plants may therefore be

defined as a group of plants that possess some special properties or virtues that

qualify them as articles of drugs and therapeutic agents, and are used for medicinal

3
purposes. Plants, including many now used as culinary herbs and spices, have been

used as medicines from prehistoric times. Spices have been used partly to counter

food spoilage bacteria, especially in hot climates, and especially in meat dishes

which spoil more readily. Angiosperms (flowering plants) were the original source

of most plant medicines. Human settlements are often surrounded by weeds useful

as medicines, such as nettle, dandelion and chickweed. Some animals such as non-

human primates, monarch butterflies and sheep ingest medicinal plants to treat

illness. Many of the plants could be used as stimulants, poisons, hallucinogens or as

medicine because of the presence of unique or rich biological – active plant

chemicals that make a plant valuable as medicinal plant are-

1. Alkaloids (compounds have addictive or pain killing or poisonous effects

and sometimes help in important cures,

2. Glycosides (use as heart stimulant or drastic purgative or better sexual

health),

3. Tannins (used for gastro-intestinal problem like diarrhea, ulcer and for

wounds and skin diseases)

4. Volatile/essential oils (enhance appetite and facilitate digestion or use as

antiseptic/ insecticide and insect repellant properties)

5. Fixed oils (present in seeds and fruits could diminish gastric/acidity)

6. Gum-resins and mucilage (possess analgesic property that suppress

inflammation and protect affected tissues against further injury and cause

mild purgative), and

7. Vitamins and minerals (Fruits and vegetables are the sources of vitamins and

minerals and these are used properly in herbals).

4
1.5 Exploration of Medicinal Properties of Plants
The exploration of medicinal properties of plants throughout the ages was

accomplished principally through careful observation, trial and error, and accidental

discovery. Observation of animals’ instinctive discrimination between toxic and

palatable plants might also have helped primitive man in choosing those plants

which are beneficial from nutritive and medicinal standpoints. And in this process,

the human race, over the countries, has created a vast heritage of knowledge and

experience on medicinal plants in different cultures and civilizations. Most of such

indigenous knowledge was handed down, through the ages, by at first orally and

later in written form as papyri, baked clay tablets, parchments, manuscripts, herbal

and finally printed herbals, pharmacopoeias and other works.

1.6 Contribution of medicinal plants in modern medicine

It is estimated that 80% of the people in Pakistan depend on plants to cure

themselves, a 40% in China. In technologically advanced countries as the United

States, it is estimated that 60% of the population use medicinal plants habitually to

fight certain ailments. In Japan there is more demand of medicinal plants than of

ʺofficialʺ medicines. Modern medicine, through clinical tests, has been able to

validate those plants that the tradition had used with the method of test and error.

Many turned out to be been worth; others demonstrated to be innocuous; others,

potentially dangerous. Biochemical tests have been the ones that determined the

main components of the medicinal plants – the active principles. Few medicinal

plants with their active compounds and therapeutic applications have tabulated in

Table 1.1.

Table 1.1: Plant derived medicinal substance occurring used in modern medicine

5
 Drug/Chemical Action/Clinical Use Plant Source

compound

Acetyldigoxin Cardiotonic Digitalis lanata

Adoniside Cardiotonic Adonis vernalis

Aescin Anti-inflammatory Aesculus

hippocastanum

Aesculetin Anti-dysentery Frazinus rhychophylla

Agrimophol Anthelmintic Agrimonia supatoria

Ajmalicine Circulatory Disorders Rauvolfia sepentina

Allylisothiocyanate Rubefacient Brassica nigra

Anabesine Skeletal muscle, Anabasis sphylla

relaxant

Andrographolide Baccillary dysentery Andrographis

paniculata

Anisodamine Anticholinergic Anisodu stanguticus

Anisodine Anticholinergic Anisodus tanguticus

Arecoline Anthelmintic Areca catechu

Asiaticoside Vulnerary Centellaasiatica

Atropine Anticholinergic Atropa belladonna

Benzyl benzoate Scabicide Oligoneuron rigidum

Betulinic acid Anticancerous Betula alba

Borneol Antipyretic, Cinnamomun burmannii

analgesic, anti-

6
 inflammatory

Bromelain Anti-inflammatory, Ananas comosus

proteolytic

Caffeine CNS stimulant Camellia sinensis

Camphor Rubefacient Cinnamomum

camphora

Camptothecin Anticancerous Camptotheca acuminata

(+)-Catechin Haemostatic Potentilla fragarioides

Chymopapain Proteolytic, mucolytic Carica papaya

Cissampeline Skeletal muscle Cissampelos pareira

relaxant

Cocaine Local anaesthetic Erythroxylum coca

Codeine Analgesic, antitussive Papaver somniferum

Colchiceine amide Antitumor agent Colchicum autumnale

Colchicine Antitumor agent, anti- Colchicum autumnale

gout

Convallatoxin Cardiotonic Convallaria majalis

Curcumin Choleretic Curcuma longa

Cynarin Choleretic Cynaras colymus

Danthron Laxative Cassia species

Deslanoside Cardiotonic Digitalis lanata

Digitalin Cardiotonic Digitalis purpurea

Digitoxin Cardiotonic Digitalis purpurea

7
 Digoxin Cardiotonic Digitalis purpurea

Emetine Amoebicide, emetic Cephaelis ipecacuanha

Ephedrine Sympathomimetic, Ephedra sinica

antihistamine

Etoposide Antitumor agent Podophyllum peltatum

Galanthamine Cholinesterase Lycoris squamigera

inhibitor

Gitalin Cardiotonic Digitalis purpurea

Glaucarubin Amoebicide Simarouba glauca

Glaucine Antitussive Glaucium flavum

Glasiovine Antidepressant Octea glaziovii

Glycyrrhizin Sweetener, Addison's Glycyrrhiza glabra

disease

Gossypol Male contraceptive Gossypium species

Hemsleyadin Bacillary dysentery Hemsleya amabilis

Hesperidin Capillary fragility Citrus species

Hydrastine Hemostatic, Hydrastis Canadensis

astringent

Kaibicacud Ascaricide Digenea simplex

Irinotecan Anticancer, antitumor Camptotheca acuminata

agent

Kawain Tranquillizer Piper methysticum

Kheltin Bronchodilator Ammi visage

8
Lanatosides A, B, C Cardiotonic Digitalis lanata

Lapachol Anticancer, antitumor Tabebuia sp.

a-Lobeline Smoking deterrant, Lobelia inflate

respiratory stimulant

Menthol Rubefacient Mentha species

Methyl salicylate Rubefacient Gaultheria procumbens

Monocrotaline Antitumor agent Crotalaria sessiliflora

(topical)

Morphine Analgesic Papaver somniferum

Neoandrographolide Dysentery Andrographis

paniculata

Nicotine Insecticide Nicotiana tabacum

Nordihydroguaiaretic Antioxidant Larrea divaricata

acid

Noscapine Antitussive Papaver somniferum

Ouabain Cardiotonic Strophanthus gratus

Pachycarpine Oxytocic Sophora pschycarpa

Palmatine Antipyretic, Coptis japonica

detoxicant

Physostigmine Cholinesterase Physostigma venenosum

Inhibitor

Papain Proteolytic, mucolytic Carica papaya

Papavarine Smooth muscle Papaver somniferum

9
 relaxant

Phyllodulcin Sweetner Hydrangea serrata

Picrotoxin Analeptic Anamirta cocculus

Pilocarpine Parasympathomimetic Pilocarpu jaborandi

Pinitol Expectorant Pisonia grandis

Podophyllotoxin Antitumor anticancer Podophyllum peltatum

agent

Protoveratrines A, B Antihypertensives Veratrum album

Pseudoephredrine Sympathomimetic Ephedra sinica

Pseudoephedrine, nor- Sympathomimetic Ephedra sinica

Quinidine Antiarrhythmic Cinchona ledgeriana

Quinine Antimalarial, Cinchona ledgeriana

antipyretic

Qulsqualic acid Anthelmintic Quisqualis indica

Rescinnamine Antihypertensive, Rauvolfia serpentine

tranquillizer

Reserpine Antihypertensive, Rauvolfia serpentine

tranquillizer

Rhomitoxin Antihypertensive, Rhododendron molle

tranquillizer

Rorifone Antitussive Rorippa indica

Rotenone Piscicide, Insecticide Onchocarpus nicou

Sanguinarine Dental plaque Sanguinaria

10
 inhibitor Canadensis

Rotundine Analagesic, sedative, Stephania sinica

traquillizer

Rutin Capillary fragility Citrus species

Salicin Analgesic Salix alba

Santonin Ascaricide Artemisia maritma

Scillarin A Cardiotonic Urginea maritime

Scopolamine Sedative Datura species

Sennosides A, B Laxative Cassia species

Silymarin Antihepatotoxic Silybum marianum

Sparteine Oxytocic Cytisus scoparius

Stevioside Sweetner Stevia rebaudiana

Strychnine CNS stimulant Strychnosnux vomica

Taxol Antitumor agent Taxus brevifolia

Teniposide Antitumor agent Podophyllum peltatum

1.7 Phytotherapy- development of new synthetic drug

Phytotherapy, the treatment of disease by the use of plants, was the beginning of

pharmacotherapy or treatment of disease by means of drugs. Therapeutic uses of

plants had in effect stored at the very beginning of human life on earth when the

primitive man, out of necessity and by intuition, resorted to using plants to alleviate

his sufferings from injuries and diseases. The medicinal plants have been used in

traditional medicine for hundreds of years with reputation as efficacious remedies

although three may not sufficient scientific data to substantiate their efficacy. Of

11
these, surprisingly large number are still of importance in modern medicine. In this

way, phytotherapy laid the foundation stone of all forms of medical treatment that

are practiced today (Ghani, 1998). With the development of human civilization, the

implementation of phytotherapy exhibits a stepwise development, which can be

enumerated as -

 1st stage: Crude drugs were employed, prepared in the roughest manner,

such as powdered willow leaves in the management of pain.

 2nd stage: Crude drugs were converted into more active and manageable

forms, such as extracts or solutions, watery or alcoholic.

 3rd stage: The pure active principles separated from the crude drug were

employed, example - salicylic acid.

 4th stage: Attempt to synthesize the active drug in the laboratory and indeed

structural modification, example - aspirin, the wonder drug.

1.8 Medicinal Plants of Bangladesh

South Asian countries have a large number of naturally growing valuable medicinal

plants naturally growing. In Bangladesh 5,000 species of angiosperm are reported to

occur (IUCN, 2003). The number of medicinal plants included in the materia

medica of traditional medicine in this subcontinent at present stands at about 2,000

and more than five hundred of such medicinal plants has so far been enlisted as

growing in Bangladesh. The major geographical locations in Bangladesh that are

rich in medicinal plants have been shown in Figure 1.2.

12
 Figure 1.2: Major medicinal plants production zones in Bangladesh

1.8.1 Medicinal plants, its cultivation and Bangladeshi market

The Bangladeshi herbal medicine market is valued at Tk. 3,300 million

(approximately US$ 60 million) at trade prices. The turnover figures in 2003 were

Tk. 1,000 million, Tk. 1,800 million and Tk. 500 million for ayurvedic, unani and

homeopathy respectively. Medicinal plants though play an important role in

healthcare systems of Bangladesh, unfortunately, there is almost no report of

cultivation of medicinal plants in our country. A study reported some of the

cultivation of medicinal plants in Rajshahi division (Hussain et al, 2004). The

largest patch of medicinal plant cultivation was reported first from Laxmipur union

13
of Natore sadar upazilla in the daily newspaper Protom Alo. The medicinal plant

beparis (middleman) of Natore reported the cultivation of ulatkambalat. The

ayurvedic companies also reported about some sporadic farming of medicinal plants

in different areas of Bangladesh. One of the ayurvedic company reported that some

of their supplied raw materials like badarlathi, ulatkambal, Sharnalataare supplied

from Modhupur, Satkhira which is cultivated there by a small group of farmers.

Government formed a cell for medicinal plant in the ‘Ministry of Environment and

Forest’ to work on-A research center for medicinal plant,

a) Promoting implantation,

b) Medicinal plant seedling production through tissue culture, and

c) Leasing of lands for medicinal plant cultivation.

At the same time, some initiatives are also taken by the Department of Forestry.

Fifty-seven different medicinal plant varieties are planted in the adjacent area of

Salna national park, Gazipur. In 2001-02 financial year, it was only on 2.02 acres

land. Later in the year, they extended it to 35 acres land. The government is also

selling different medicinal plants at a subsidized rate to 400 different government

nurseries all over the country. There are also 450 sub-centers in each upazila under

the 400 government nurseries. There are some private companies like ‘Gemcon

Food Products’ or ‘Nim Foundation’ who have farms of medicinal plants. ‘Gemcon

Food Products’ are preparing some herbal medicine in their cottage industry which

are available in the market. The farms are at Dinajpur. The ‘Nim foundation’ also

has a medicinal farm at Faridpur district. They planted mainly nim plant to produce

beauty products. The whole procedure starts from phytochemistry, where plants

materials are extracted and experimented for their medicinal use.

14
1.9 Medicinal Plants: Wealth of a Country
Medicinal plants constitute and important natural wealth of a country. They play a

significant role in providing primary health care services to rural people. They serve

as important therapeutic agents as well as important raw materials for the

manufacture of traditional and modern medicines. Medicinal plants are rich sources

of bioactive compounds and thus serve as important raw material for drug

production. It has now been established that the plants synthesize and accumulate

some secondary metabolites like alkaloids, glycosides, tannins, volatile oils etc. that

may possess a great potential for biological activity and can be a curative agent in

therapeutic purposes. As therapeutic uses of plants continued with the progress of

civilization and development of human knowledge, scientists endeavored to isolate

different chemical constituents from plants, put them to biological and

pharmacological tests and thus have been able to identify and isolate therapeutically

active compounds. The 19th century saw the scientific revolution in medicine. The

first isolation and crystallization of an active drug from a natural source was the

accomplishment of a pharmacist’s assistant, Sertuner (1783-1841), who obtained

pure morphine from natural opium in 1803. Pure quinine was isolated from

cinchona Leaves in 1820 (Goldstein, Aronow et al. 1990). Isolation of other

important plant derived drugs of modern medicine rapidly followed and many

useful drugs have since been discovered and introduced into modern medicine.

Drugs like strychnine from Strychnosnux-vomeca (1817), caffeine from

Theasinensis (1819), quinine from Chcinchona spp. (1820) and colchicines from

Colchicum autumnale (1820) constitutes some example of such early drugs (Ghani

1998). Facilitated by the rapid development of technology of isolation and

characterization process that is chromatographic and spectroscopic methods, a large

15
number of therapeutically active plant constituents have been isolated during last

two decades. In the later part of the 19th century the German dye industry led the

way toward the deliberate synthesis of new drugs and the molecular modification of

existing ones. Still now the medicinal chemistshave a great interest in the molecular

modification of therapeutic agent isolated from plant source. For example, taxotere

that enjoys an extensive use in present cancer chemotherapy is thesynthetic analog

of paclitaxel, developed by the scientists of Rhone Poulenc Rorer (Goldstein,

Aronow et al. 1990).

1.10 Plant Kingdom: Storehouse of Innumerable Drugs

Plant kingdom, the storehouse of thousands of unexplored compounds, possesses a

great potentiality for drug search even in the day of synthetic chemistry. The

following data shows how the plant kingdom enriches the modern medicinal

practice. About 33% of the drugs produced in the developed countries are derived

from plants (Goldstein, Aronow et al. 1990). An analysis of over 300 million

prescriptions for the year 1960 revealed that 47 percent were for drugs of natural

origin, mostly antibiotics (Gossehn 1962). In the United States, in 1980 alone, the

consumer paid 8 billion dollars for prescription drugs in which the active

ingredients are still derived from plants. If microbes are added, 60% of the modern

medicinal products are of natural origin (Goldstein, Aronow et al. 1990). More than

47% of all drugs used in Russia are obtained from botanical sources. According to

some generous estimates, almost 80% of the present day medicines are directly or

indirectly obtained from plants (Katiyar, Gupta et al. 2012).

16
1.11 Modern Prescription Drugs
Many of the remedies employed by the herbalists provided effective treatments.

Studies of foxglove for the treatment of dropsy (congestive heart failure) set the

standard for pharmaceutical chemistry. In the 19th century, scientists began

purifying the active extracts from medicinal plants (e.g., the isolation of morphine

from the opium poppy). Advances in the field of pharmacology led to the

formulation of the first purely synthetic drugs based on natural products in the

middle of the 19th century. In 1839, for example, salicylic acid was identified as the

active ingredient in a number of plants known for their pain-relieving qualities;

salicylic acid was synthesized in 1853, eventually leading to the development of

aspirin. It is estimated that 25% of prescriptions written in the U.S. contain plant-

derived ingredients (close to 50% if fungal products are included); an even greater

percentage are based on semisynthetic or wholly synthetic ingredients originally

isolated from plants (Katiyar, Gupta et al. 2012).

1.11.1 Herbal Medicine Today

While Western medicine strayed away from herbalism, 75% to 90% of the rural

population of the rest world still relies on herbal medicine as their only health care.

In many village marketplaces, medicinal herbs are sold alongside vegetables and

other wares. The People’s Republic of China is the leading country for

incorporating traditional herbal medicine into a modern health care system; the

result is a blend of herbal medicine, acupuncture, and Western medicine. Plantations

exist in China for the cultivation of medicinal plants, and thousands of species are

thus available for the Chinese herbalist; prescriptions are filled with measured

amounts of specific herbs rather than with pills or ointments. In India, traditional

17
systems have remained quite separate from Western medicine. In addition to

Ayurvedic medicine, which has a Hindu origin, Unani medicine, with its Muslim

and Greek roots, is another widely practiced herbal tradition in India. The renewed

interest in medicinal plants has focused on herbal cures among indigenous

populations around the world, especially those in the tropical rain forests. It is

hoped that these investigations will add new medicinal plants to the world’s

pharmacopoeia before they are lost forever. In addition to the destruction of the

forests, the erosion of tribal cultures is also a threat to herbal practices (Dangol

2008).

1.11.2 Herbal Drug Research: Bioactivity Guided Approach

Plants, the molecular architect still offers a great potentiality for drug search. Plants

contain compounds having interesting skeletons. Bioactivity guided phytochemical

investigation of medicinal plants may yield newer chemical constituents of

remarkable therapeutic interest. Occasionally, native lore provides clue to plants

with pharmacologic activity. Digitalis, opiates and the cinchona alkaloids (quinine

and quinidine) came into modern medicine by this route. Curare was obtained from

a South American plant long used by natives to prepare arrow poison.

Rawolfiaserpentina was used for a variety of illnesses; only in recent years its

transquilizing properties were recognized in Western medicine and its active

principle, reserpine, were isolated (Goldstein, 1974). Books on herbal medicinal

practice report numerous medicinal plants, which are still not investigated. These

plants can be subjected to pharmacologic screening as per their traditional use to

evaluate their utility. In case of significant result, chromatographic andspectroscopic

18
methods can be applied to isolate the responsible agent. Bioactivity guided

approach has three characteristic phases of investigation: -

 Biological activity is detected in crude material, and a bioassay system is set

up to permit the identification of active fractions and discarding the inactive

ones.

 The crude material is fractionated by the most appropriate chemical

procedures, all fractions are tested, and active fractions are further

fractionated, and so on, until pure compounds are obtained.

 The chemical structures of pure compounds are determined (Goldstein,

Aronow et al. 1990).

1.12 Research in Medicinal Plant

It is very important to ensure that these medicinal plants or their products really

possess the claimed properties and exert the desired therapeutic effects. A

phytochemist uncovering these resources is producing useful materials for screening

programs for drug discovery. Emergence of newer disease also leading the scientists

to go back to nature for newer effective molecules. Recently developed genetic

engineering in plants has increased their importance in the field of medicine. In

order to substantiate the above assertion about their validity these medicinal plants

must be subjected to extensive scientific study (Ghani 1998). Medicinal plants

constitute the major constituents of indigenous medicines. Thus, the quality and

effectiveness of medicinal preparations depend solely in case of indigenous

medicines, on the genuineness and quality of the medicinal plants or products that

are used in their preparation. So it is very important to ensure that these medicinal

plants or their products really possess the claimed properties and exert the desired

19
therapeutic effects. But there is hardly any medicinal plant which is used forasingle

therapeutic purpose. Again, all medicinal plants do not really possess the claimed

therapeutic virtues and medicinal properties, because most of these claims are based

on old literature, folk sayings, occasional experiences and traditional uses, but not

on any significant clinical or pharmacological studies and statistical data. Thus the

claims of medicinal properties may not be true or scientifically valid in case of all

the so called medicinal plants (Ghani 1998). A phytochemist uncovering these

resources is producing useful materials for screening programs for drug discovery.

Emergence of newer disease also leading the scientists to go back to nature for

newer effective molecules. Recently developed genetic engineering in plants has

further increased their importance in the field of medicine, for example in the

production of antibiotics by expression of an appropriate gene in the plant. By using

these it is possible to modify the activity or regulate the properties of the key

enzymes responsible for the production of secondary metabolites. Thus, by knowing

the potential resources it is possible to increase the content of the important active

compounds and in the future, genes responsible for very specific biosynthetic

processes may be encoded into host organism to facilitate, difficult synthetic

transformation. Thus, plants are considered as are of the most important and

interesting subjects that should be explored for the discovery and development of

newer and safer drug candidates. In order to substantiate the above assertion about

their validity these medicinal plants must be subjected to extensive scientific study.

Attempts must be made to separate the real medicinal plants from the useless ones,

and for this, extensive phytochemical and pharmacological investigations must be

carried out on these plants. This will help in making the indigenous systems more

scientific and more reliable. At the same time much of the superstitions, wrong

20
 impressions and over-estimations associated with the medicinal plants will be

removed (Ghani 1998).

1.12.1 Rationale for Herbal Drug Research in Bangladesh

The number of plants with medicinal properties included in the traditional medicine

in this subcontinent as present stands at about 2000. More than 500 of such

medicinal plants have so far been established as growing in Bangladesh. This

number of the indigenous medicinal plants is in the increase with the discovery and

introduction of newer plants every day (Ghani 1998). Almost all these indigenous

medicinal plants are extensively used in the preparation of unani, ayurvedic and

homeopathic medicines in Bangladesh. But the fact is that almost all of them are

lacking scientific evaluation of their therapeutic uses. So standard pharmacological

and phytochemical methods should be applied in order to, establish scientific

evidence of their pharmacological use and isolate the active constituent. This may

offer a local natural source of a commonly used drug or a novel therapeutic agent.

1.13 Some plants with their traditional uses

Table 1.2: Some medicinal plants with their traditional uses

Scientific name Family Life form Part used and uses

Annona reticulata L. Annonaceae Tree Leaf paste is applied on the

affected area of scabies.

Polyalthia longifolia Annonaceae Tree Stem bark used as febrifuge

Cocculus hirsutus L. Menispermaceae Climber Leaf juice is applied on the

affected areas of eczema till

cure.

21
Cadaba fruticosa L. Capparaceae Shrub Root decoction is

administered during

helminthiasis

Hybanthus Violaceae Herb Roots are diuretic and given

enneaspermus in the urinary disorders

Cochlospermum Cochlospermace Tree Stem bark paste is applied

religiosum L. ae over the bone fractured areas

Casearia elliptica Flacourtiaceae Tree Leaf paste is applied over the

bone fractured areas

Flacourtia indica Flacourtiaceae Shrub Leaf juice is given internally

to treat jaundice till cure

Urena lobate L. Malvaceae Shrub Roots are diuretic

Waltheria indica L. Sterculiaceae Herb Roots chewed to control

internal haemorrhages

Grewia tiliaefolia Tiliaceae Tree Stem bark is used in

dysentery

Triumfetta rhomboidea Tiliaceae Herb Leaf paste is applied on the

affected areas of scabies and

eczema

Aegle marmelos L. Rutaceae Tree Half ripe fruits used for

diarrhea and dysentery

Atlantia monophylla L. Rutaceae Tree Oil from the fruit used in

Rheumatism

22
Chloroxylon swietenia Rutaceae Tree Leaves used in Rheumatism

Glycosmis pentaphylla Rutaceae Shrub Juice of leaves used in fever,

liver complaints and other

skin troubles

Toddalia asiatica L. Rutaceae Shrub Root bark is diaphoretic and

used in stomach ache.

Azadirachta indica Meliaceae Tree Stem bark used for skin

troubles

Cissus quadrangularis L. Vitaceae Climber Stem paste used to cure

rheumatoid arthritis

Sapindus emarginatus Sapindaceae Tree Fruits useful in treating

asthma,diarrhea

Schleichera oleosa L. Sapindaceae Tree Seed oil is used for massage

in rheumatism

Lannea coromandelica Anacardiaceae Tree Stem bark used as astringent

Mangifera indica L. Anacardiaceae Tree Stem bark is decoction is

administered for diarrhea

Semecarpus anacardium Anacardiaceae Tree Gum used in leprosy and

nervous debility.

Abrus precatorius L. Fabaceae Climber Root decoction used for

coughs and cold, diuretic.

Butea superba Roxb. Fabaceae Climber Decoction of shoots used in

piles

23
Cajanus cajan L. Fabaceae Shrub Leaf extract is administered

for stomach pain

Canavalia gladiata Fabaceae Climber Leaf juice used in case of

abdominal pains

Desmodium gangeticum Fabaceae Herb Root paste used in whooping

cough

Desmodium triflorum Fabaceae Herb Leaves used for dysentery and

diarrhea

Mucuna monosperma Fabaceae Climber Seeds used in cough, asthma

Tephrosia purpurea L. Fabaceae Herb Decoction of roots given in

diarrhea, rheumatism and

asthma

Cassia fistula L. Fabaceae Tree Dried fruits used as laxative

Albizia Lebbeck L. Fabaceae Tree Leaf juice administered orally

to treat night blindness

Dichrostachys cinerea Mimosaceae Tree Root paste is applied during

bone fracture

Xylea xylocarpa Mimosaceae Tree Root bark paste is applied on

the affected areas of syphilis

Kalanchoe lanceolata Crassulaceae Herb Leaf juice given during

dysentery

Tamarindus indica L. Fabaceae Tree Fruit pulp is used as a laxative

Calycopteris floribunda Combretaceae Climber Leaf juice used as laxative

24
Quisqualis indica L. Combretaceae Shrub Seeds used for treating

helmenthiasis

Terminalia bellarica Combretaceae Tree Decoction of fruits is

administered for

haemorrhoids

Woodfordia fruticosa L. Lythraceae Shrub Flowers used in menorrhagia

Coccinia grandis L. Cucurbitaceae Climber Leaf paste applied on the

affected areas of scabies

Opuntia dillenii Cactaceae Shrub Paste of phyllode and stem

bark is applied on the area of

snake bite

Alangium salvifolium Alangiaceae Tree Root bark is applied for skin

troubles

1.14 Drug Preparation Based on Medicinal Plant Study

Although there is much research in molecular modeling, combinatorial chemistry,

and other synthetic chemistry techniques which has been funding by pharmaceutical

companies and organizations, natural products which have much complicated

structural formulas and particularly medicinal plants, remain an important source of

new drugs, new chemical entities (NCEs) and new drug leads (Newman, Cragg et

al. 2003, Butler 2004). According to survey in 2001 and 2002, approximately one

quarter of the best-selling drugs in the world were natural products or derived from

natural products (Butler 2004). It has also been reported that approximately 28% of

NCEs between 1981 and 2002 were natural products or natural product-derived

natural products (Newman, Cragg et al. 2003)and another survey during this period

25
20% of NCEs were considered natural product mimics, meaning that the synthetic

compound was derived from the study of natural products (Newman, Cragg et al.

2003). On the bases of this report it has been assumed that research on natural

products accounts for approximately 48% of the NCEs reported from 1981–2.

1.15 Drug Development from Plant Source

Development of drugs from medicinal plants is often an elaborate, laborious, time-

consuming and expensive exercise. Careful phytochemical analysis and

pharmacological and clinical tests are pre-requisites for developing drugs from

medicinal plants. Modern drug development from plant source is carried out

according to a systemic investigation as described.

 Selection of the proper medicinal plant.

 Appropriate consolidation of medicinal plant.

 Extraction with suitable solvent(s).

 Detection of biological activity of crude extract and establishment of a

bioassay system to permit the identification of the active fractions and

rejection of the inactive ones.

 Fractionation of crude extracts using the most appropriate chromatographic

procedures, biological evaluation of all fractions and separation of the active

fractions.

 Repeated fractionation of active fractions to isolate pure compound(s).

 Evaluation of biological activity of pure compound(s).

 Investigation of other pharmacological properties of the active agent(s).

 Toxicological tests with pure compound(s).

 Production of drug in appropriate dosage forms.

26
There have no pharmacological studies have been performed to assess the

Hepatoprotective & Hypoglycemic action of this species as a folk medicine. This

study deals with the pharmacological actions namely Hepatoprotective &

Hypoglycemic effects of a newer source of indigenous medicinal plant Fimbristylis

miliacea.

27
 Chapter 2: Literature review
2.1 Introduction to the Plant Family: Cyperaceae
Cyperaceae is one of the most widely distributed plant groups. It was formally

described by De Jussieu in 1789; the name is derived from the genus name Cyperus,

originally from the Greek kupeiros, meaning sedge. It is the seventh largest family

in the angiosperms and third largest family in the Monocotyledons after Poaceae

and Orchidaceae. Worldwide 109 genera with 5500 species of the family were

reported (Govaerts et al., 2007), of which 38 genera with about 485 species are

known to grow in India (Karthikeyan et al., 1989). The Cyperaceae are grass like

herbaceous plants found especially in wet regions throughout the world. Cyperaceae

members are always herbs except the African genus Microdracoides which is tree -

like. Sedges are characterized by the grass -like or rush -like habit with or without

rhizomes or stolons, the minute, inconspicuous flowers enclosed by the distichously

or spirally arranged glumes on a spikelet and the indehiscent fruits known as nuts or

achenes.

Sedges are usually annual or perennial herbs. Annuals are with fibrous roots and

perennials with short or long creeping rhizomes. The rhizomes are usually small,

woody, but sometimes are long creeping or emitting stolons which often bear tubers

as Bolboschoenus maritimes (Linnaeus) Palla and Cyperus rotundus (L.). The

rhizomes and stolons are clothed with scales which usually disintegrate leaving

fibrous remains.

2.1.1 Morphology

 The stems of Cyperaceae are often triangular and mostly solid.

28
  Spikelets of Cyperaceae the individual flowers are subtended by a single

scale.

 Fibrous roots.

 The flowers are very small.

 The flowers are arranged along a shortened axis, called a rachilla.

 Fruits are achenes.

2.1.2 Habitat

In general, Cyperaceae members are found in varied habitats. There are certain

genera confined to particular habitats. For example, species of Carex are generally

found in hilly forest areas. Cyperus one of the most dominant genera in Southern

Assam is found in different habitats but mostly in lowland, wet or marshy areas.

Many species, especially of Cyperus, Fimbristylis, Mariscus, Eleocharis etc. are

found in weeds in agricultural fields. Dispersal of sedges very much depends on the

habitat of the species. Species growing in and around water is mostly dispersed

through water only. Species found in and around fresh water bodies like marshes,

swampy areas, rice fields and other wet land or along riverbanks, lakes etc. are also

dispersed through water. Most of these nuts sink in water and are often carried by

flowing water or may survive till the water dries up in the seasonal water bodies.

There are few species in which some devices are found for their dispersal. In Carex,

the nut is enclosed by a sac like utricle, which floats on water surface and is

dispersed by water current. Nut of many sedges found as weeds are brought to

agricultural lands, especially rice fields through irrigation water or along with the

crop seeds. Tubers and stolons of some species like Cypenes rotundus are also

dispersed through irrigation water.

29
2.1.3 Genera of the Cyperaceae Family

Cyperaceae family consists of 110 plant genera. The Plant List includes 18,812

scientific plant names of species rank for the family Cyperaceae. of these 5,784 are

species names. It includes:

Abildgaardia, Acriulus, Actinoschoenus, Actinoscirpus, Afrotrilepis, Alinula,

Amphiscirpus, Androtrichum, Arthrostylis, Ascolepis, Baumea, Becquerelia,

Bisboeckelera, Blepharolepis, Blysmopsis, Blysmus Bolboschoenoplectus,

Bolboschoenus, Bulbostylis, Calyptrocarya, Capeobolus, Capitularina,

Carex, Carpha, Caustis, Cephalocarpus, Chaetocyperus, Chaetospora,

Chorizandra, Chrysitrix, Cladium, Cobresia, Coleochloa, Costularia,

Courtoisina, Crosslandia, Cyathochaeta, Cyathocoma, Cymophyllus,

Cyperus, Cypringlea, Dichostylis, Dichromena, Diplacrum, Diplasia,

Dulichium, Ecklonea, Eleocharis, Elynanthus, Epischoenus, Eriophorum,

Eriospora, Evandra, Everardia, Exocarya, Ficinia, Fimbristylis, Fuirena,

Gahnia, Gymnoschoenus, Heleocharis, Hellmuthia, Holoschoenus, Hoppia,

Hymenolytrum, Hypolytrum, Isolepis, Juncellus, Karinia, Khaosokia,

Kobresia, Koyamaea, Kyllinga, Kyllingiella, Lagenocarpus, Lepidosperma,

Lepironia, Limnochloa, Lipocarpha, Loxotrema, Machaerina, Mapania,

Mariscus, Melancranis, Mesomelaena, Microdracoides, Morelotia,

Neesenbeckia, Nelmesia, Nemum, Neoscirpus, Neskiza, Oncostylis,

Oreobolopsis, Oreobolus, Oxycaryum, Paramapania, Phylloscirpus,

Pleurostachys, Principina, Pseudoschoenus, Psilocarya, Ptilothrix, Pycreus,

Queenslandiella, Reedia, Remirea, Rhynchocladium, Rhynchospora,

Schoenoplectiella, Schoenoplectus, Schoenoxiphium, Schoenus,

30
 Scirpodendron, Scirpoides, Scirpus, Scleria, Sphaerocyperus, Stenophyllus,

Sumatroscirpus, Syntrinema, Tetraria, Thoracostachyum, Torulinium,

Trachystylis, Trianoptiles, Trichophorum, Trichoschoenus, Tricostularia,

Trilepis, Uncinia, Vignea, Vincentia, Volkiella, Zameioscirpus.

2.1.4 The plant genus Fribristylis

Fimbristylis is a genus of sedges. A plant in this genus may be known commonly as

a fimbry, fimbristyle, or fringe-rush. There are 200 to 300 species distributed

worldwide. Several continents have native species but many species have been

introduced to regions where they are not native. Some are considered weeds. These

are typical sedges in appearance, with stiff, ridged stems and cone-shaped terminal

panicles of spikelets. They are found in wet environments, and are most diverse in

tropical and subtropical regions.

2.1.5 Species: Members of the genus Fimbristylis

The Plant List for the genus Fimbristylis taken from the version 1.1 (Published on

the internet), 2012. Retrieved December 16, 2018. [12]

Fimbristylis acicularis R.Br., F. acuminata Vahl, F. adenolepis J.Kern, F. adjuncta

S.T.Blake, F. adventitia Ces., F. aestivalis Vahl, F. aggregate C.E.C.Fisch., F.
alata E.G.Camus, F. albicans Nees, F. albovirens C.B. Clarke, F. alboviridis
C.B.Clarke, F. ambavanensis V.P.Prasad & N.P.Singh, F. ammobia Latz, F.
amplocarpa Govind., F. angamoozhiensis Ravi & Anil Kumar, F. anisoclada Ohwi,
F. aphylla Steud., F. argentea (Rottb.) Vahl, F. argillicola Kral, F. arnhemensis
Latz, F. arnottiana Boeckeler, F. arthrostyloides W.Fitzg., F. aspera (Schrad.)
Boeckeler, F. assamica C.B.Clarke ex Guha Bakshi, F. autumnalis (L.) Roem. &
Schult., F. bahiensis Steud., F. barteri Boeckeler, F. benthamiana Govind., F.
bispicula Govind., F. bisumbellata (Forssk.) Bubani, F. bivalvis (Lam.) Lye, F.
blakei Latz, F. blepharolepis J.Kern, F. borbonica Steud., F. breviculma Govind.,
F. brevivaginata Kral, F. brunneoides J.Kern, F. caesia Miq., F. caespitosa R.Br.,

31
F. calcicole J.Kern, F. caloptera Latz, F. cancellata Cherm., F. capilliculmis Ohwi,
F. cardiocarpa F.Muell., F. caroliniana (Lam.) Fernald, F. carolinii Latz,
F. carpopoda Govind., F. celebica Ohwi, F. cephalophora F.Muell.,
F. cephalotes Steud., F. chingmaiensis S.M.Huang, F. cinnamometorum (Vahl)
Kunth, F. circumciliata Govind., F. clavata S.T.Blake, F. compacta Turrill,
F. complanata (Retz.) Link, F. composita Latz, F. consanguinea Kunth,
F. contorta C.E.C.Fisch., F. corynocarya F.Muell., F. costiglumis Domin,
F. crystallina Govind., F. cuneata Govind. & S.K.Varma, F. cymosa R.Br.,
F. dauciformis Govind., F. debilis Steud., F. decipiens Kral, F. densa S.T.Blake, F.
denudate R.Br., F. dichotoma (L.) Vahl, F. dictyocolea S.T.Blake, F. didrichsenii
Boeckeler, F. diglumoides Govind. & S.K.Varma, F. dimorphonucifera Govind.,
F. diphylloides Makino, F. dipsacea (Rottb.) C.B.Clarke, F. disticha Boeckeler,
F. distincta S.T.Blake, F. dolera S.T.Blake, F. doliiformis Govind.,
F. donticola Hochst. ex Steud., F. dunlopii Latz, F. dura (Zoll. & Moritzi) Merr.,
F. eichleriana Govind., F. elegans S.T.Blake, F. eligulata Govind.,
F. elongata Pires de Lima, F. engleriana Buscal. & Muschl., F. eragrostis (Nees)
Hance, F. eremophila Latz, F. falcata (Vahl) Kunth, F. falcifolia Boeckeler,
F. faulensis Beck, F. fenestrata Kük., F. ferruginea (L.) Vahl, F. fibrillosa Goetgh.,
F. filifolia Boeckeler, F. fimbristyloides (F.Muell.) Druce, F. fordii C.B.Clarke,
F. formosensis C.B. Clarke, F. fuchsiana Govind., F. fulvescens (Thwaites)
Thwaites, F. furva R.Br., F. fusca (Nees) Benth. ex C.B.Clarke,
F. fuscinux C.B.Clarke, F. fuscoides C.B.Clarke, F. gabonica Cherm.,
F. gambleana Boeckeler, F. gentarea Govind., F. gigantea Kük.,
F. glaucophylla (Boeckeler) Beetle, F. glazioviana Boeckeler, F. gracilenta Hance,
F. griffithii Boeckeler, F. hamiltonii Steud., F. hawaiiensis Hillebr.,
F. henryi C.B.Clarke, F. hirsutifolia Govind., F. hookeriana Boeckeler,
F. humerosa Govind., F. humilis S.T.Blake, F. hyalina Govind. & Sasidh.,
F. hygrophila Gordon-Gray, F. inaguensis Britton, F. insignis Thwaites,
F. intonsa S.T.Blake, F. × itaru-itoana T.Koyama, F. jucunda (C.B.Clarke) J.Kern,
F. juncea (G.Forst.) R.Br. ex Roem. & Schult., F. juncocephala Boeckeler,
F. kadzusana Ohwi, F. kernii T.Koyama, F. kingii Gamble ex Boeckeler,
F. kwantungensis C.B.Clarke, F. lanata Roem. & Schult., F. lanceolata C.B.Clarke,
F. lasiophylla J.Kern, F. latiglumifera Govind., F. latinucifera Govind.,

32
F. lawiana (Boeckeler) J.Kern, F. laxiglumis Latz, F. leptoclada Benth.,
F. leucocolea Benth., F. leucostachya Roem. & Schult., F. ligulata Govind.,
F. limosa Poepp. & Kunth, F. lineatisquama Ohwi, F. lithophila Govind.,
F. littoralis Gaudich., F. longibracteata Pires de Lima, F. longispica Steud.,
F. longistigmata Govind., F. longistipitata Tang & F.T.Wang,
F. macassarensis Steud., F. macrantha Boeckeler, F. madagascariensis Boeckeler,
F. magnifica C.B.Clarke, F. malayana Ohwi, F. mangorensis Cherm.,
F. manilaliana Govind., F. maracandica Zakirov, F. merguensis C.B.Clarke,
F. merrillii J.Kern, F. mexicana Palla, F. micans S.T.Blake,
F. microcarya F.Muell., F. modesta S.T.Blake, F. monospicula Govind.,
F. monticola Hochst. ex Steud., F. mozambicensis Gand., F. multicephala Govind.,
F. multinervia Govind., F. mycosa Govind., F. nagpurensis V.P.Prasad &
N.P.Singh, F. naikii Wad.Khan & Lakshmin., F. nanningensis Tang & F.T.Wang,
F. narayanii C.E.C.Fisch., F. neilsonii F.Muell., F. nelmesii J.Kern,
F. neocaledonica C.B.Clarke, F. nigrescens Steud., F. nigritana C.B.Clarke,
F. nigrobrunnea Thwaites, F. nuda Boeckeler, F. nutans (Retz.) Vahl,
F. oblonga T.Koyama, F. obtusata (C.B.Clarke) Ridl., F. odontocarpa S.T.Blake,
F. onchnidiocarpa J.Kern, F. ovata (Burm.f.) J.Kern, F. oxystachya F.Muell.,
F. pachyptera S.T.Blake, F. palauensis Ohwi, F. pallida S.T.Blake,
F. pandurata Govind., F. parvilenta T.Koyama, F. pauciflora R.Br.,
F. paupercula Boeckeler, F. pentastachya Boeckeler, F. perlaxa Ohwi,
F. perpusilla R.M.Harper. ex Small & Britton, F. perspicua Govind. & Sasidh.,
F. phaeolepis J.Kern, F. phaeoleuca S.T.Blake, F. pierotii Miq.,
F. pilifera W.Fitzg., F. pilosa Vahl, F. planifolia Steud., F. polytrichoides (Retz.)
Vahl, F. prabatensis D.A.Simpson, F. prolifera Steud., F. psammocola Tang &
F.T.Wang, F. psammophila J.Kern, F. pseudomicrocarya Govind.,
F. pseudonarayanii Ravi & Anil Kumar, F. pterygosperma R.Br., F. puberula Vahl,
F. pubisquama J.Kern, F. punctata R.Br., F. pustulosa Govind.,
F. quadriflora (Boeckeler) Beetle, F. quinquangularis (Vahl) Kunth, F. rara R.Br.,
F. ratnagirica V.P.Prasad & N.P.Singh, F. raymondii T.Koyama,
F. recta F.M.Bailey, F. rectifolia Govind., F. rhizomatosa Pires de Lima,
F. rhodesiana Rendle, F. rhyticarya F.Muell., F. rigidiuscula Govind.,
F. rigidula Nees, F. rugosa Govind., F. rupestris Latz, F. sachetiana Fosberg,

33
F. salbundia (Nees) Kunth, F. sanjappae W.Khan & Solanke,
F. savannicola J.Kern, F. scaberrima Nees, F. scabrida Schumach.,
F. scabrisquama Govind., F. schoenoides (Retz.) Vahl, F. schultzii Boeckeler, F.
schulzii Boeck., F. schweinfurthiana Boeckeler, F. semarangensis Ohwi, F.
semidisticha Govind., F. semihirsuta Boeckeler, F. sericea (Poir.) R.Br.,
F. shimadana Ohwi, F. sieboldii Miq. ex Franch. & Sav., F. signata S.T.Blake,
F. simaoensis Y.Y.Qian, F. simplex S.T.Blake, F. simpsonii V.P.Prasad &
N.P.Singh, F. simulans Latz, F. singularis Govind., F. sleumeri J.Kern, F.
smitinandii T.Koyama, F. solidifolia F.Muell., F. spadicea (L.) Vahl, F. spartium
Ham., F. sphaerocephala Benth., F. spicigera J.Kern, F. spiralis R.Br., F.
splendida C.B.Clarke, F. squarrosa Vahl, F. squarrulosa F.Muell., F.
stauntonii Debeaux & Franch., F. stenostachya S.T.Blake, F.
stigmatotecta Govind., F. stolonifera C.B.Clarke, F. straminea Turrill, F. strigose
Govind., F. striolata Napper, F. subalata J.Kern, F. subaphylla Boeckeler, F.
subaristata Benth., F. subdura Ohwi, F. subinclinata T.Koyama, F.
subtrabeculata C.B.Clarke, F. subtricephala T.Koyama, F. sumbaensis Ohwi, F.
swamyi Govind., F. tamaensis Steyerm., F. tenera Schult., F. tenuicula Boeckeler,
F. tenuinervia J.Kern, F. tetragona R.Br., F. thermalis S.Watson, F.
thomsonii Boeckeler, F. tortifolia Govind., F. trachycarya F.Muell., F.
trichocaulis C.B.Clarke, F. trichoides J.Kern, F. trichophylla Ridl., F. triflora (L.)
K.Schum. ex Engl., F. trigastrocarya F.Muell., F. tristachya R.Br., F. tumida
Govind., F. tunquinensis Boeckeler, F. turkestanica (Regel) B.Fedtsch., F.
uliginosa Hochst. ex Steud., F. ultragluma Govind., F. umbellaris (Lam.) Vahl, F.
unispicularis Govind. & Hemadri, F. urakasiana Kük., F. vagans S.T.Blake, F.
vaginata (R.Br.) Domin, F. vahlii (Lam.) Link, F. vanoverberghii Kük., F.
variegate Gordon-Gray, F. villosissima Steud., F. virella Govind., F. warmingii
(Boeckeler) Malme, F. wetarensis Ohwi, F. willdenowiana Govind., F. woodrowii
C.B.Clarke, F. xyridis R.Br., F. zatei W.Khan & D.P.Chavan, F. zeylanica
T.Koyama.

34
2.2 Investigating Plant: Fimbristylis miliacea

2.2.1.1 Plant Information

 Scientific name: Fimbristylis miliacea (L.)

 Local name: ‘Fringerus’, ‘Ghueen’ or ‘Chhoti bhoin’.

 Other scientific names -

- Fimbristylis angularis Link

- Fimbristylis benghalensis (Pers.) Roem. & Schult.

- Fimbristylis fauriei Ohwi

- Fimbristylis miliacea (L.) Vahl

- Fimbristylis mucronata Vahl

- Fimbristylis quinquangularis var. bistaminifera Tang & F.T.Wang

- Fimbristylis quinquangularis var. elata Tang & F.T.Wang

- Fimbristylis quinquangularis subsp. quinquangularis

- Fimbristylis quinquangularis var. quinquangularis

- Iria angularis (Link) Kuntze

- Iria miliacea (L.) Kuntze

- Iria quinquangularis (Vahl) Kuntze

- Isolepis angularis Schrad. ex Schult.

- Isolepis miliacea (L.) J.Presl & C.Presl

- Isolepis miliacea var. major J.Presl & C.Presl

- Isolepis pentagona (Roxb.) Schult.

- Isolepis tetragona Schult.

- Scirpus benghalensis Pers.

- Scirpus miliaceus L.

- Scirpus niloticus Blanco

35
 - Scirpus parviflorus Willd. ex Kunth

- Scirpus pentagonus Roxb.

- Scirpus plantagineus Roxb.

- Scirpus quadrangularis Thouars

- Scirpus quinquangularis Vahl

- Scirpus salbundius Buch.-Ham. ex Wall.

- Scirpus tetragonus Roxb.

- Trichelostylis angularis (Link) Nees

- Trichelostylis miliacea (L.) Nees

- Trichelostylis miliacea var. microstachya Nees

- Trichelostylis pentagona Nees

- Trichelostylis quinquangularis (Vahl) Nees

- Trichelostylis tetragona (Schult.) Nees

2.2.2 Taxonomy of Fimbristylis miliacea

Domain: Eukaryota

Kingdom: Plantae

Phylum: Angiosperms

Subphylum: Monocots

Class: Commelinids

Order: Poales

Family: Cyperaceae

Genus: Fimbristylis

Species: Fimbristylis

miliacea

36
2.2.3 Plant type

Fimbristylis miliacea, is a grass like herb with a fibrous root system. The culms are

slender, 40 to 60cm tall; their transversal section is four-angled and somewhat

flattened. Leaves are linear, threadlike and stiff, two ranked, 1.5 to 2.5mm wide, up

to 40cm long, basal leaves are half as long as culm. The leaf bract is shorter than

inflorescence. Fruit of the plant is achene white, yellowish, less than half the length

of the glume, three angled, biconvex, broadest above the middle, very finely warty,

somewhat sugarcoated.

2.2.4 Morphology

 Stem is occupied by an array of spikelets, each borne on a long peduncle.

 The spikelet is spherical to ovate and reddish brown in color.

 The spikelets flower and then develop tiny fruits, which are brown achenes.

2.2.5 Distribution

The species is found throughout many countries with a tropical or sub-tropical

climate in southern and south-east Asia including: Australia, Bangladesh, Bhutan,

Cambodia, India, Indonesia, Malaysia, Myanmar, Nepal, Pakistan, Philippines, Sri

Lanka, Thailand and Vietnam. It has also been introduced into Ecuador,

Madagascar, Nicaragua, Peru and Suriname. In Bangladesh, this plant is mainly

found in Manikgonj, Rangpur, Dinajpur, Nababgonj.

2.2.6 Habitat

Paddy fields, common in moist to swampy places or in shallow water, along ditches,

drains and streams, a common weed of the paddy fields.

2.2.7 Traditional use of Fimbristylis

 Fimbristylis squarrosa Vahl; Whole plant (juice) for sore throat.

37
  Fimbristylis falcata (Vahl) Kunth; Rhizome (juice) in dysentery.

 Fimbristylis ovata (Burm.f.) J.Kern; Culms with flower (paste) in

rheumatism.

 Fimbristylis miliacea (L.) Vahl; gumi Leaves (juice) as a form of poultice to

treat fever.

2.2.8 Literature survey of Fimbristylis miliacea

No published results are available in the literature reporting phytochemical

investigation and pharmacological activities of this plant

38
 Chapter 3: Preparation of plant extract
3.1 Methods for Extract Preparation
The plant parts selected for present work were full part of Fimbristylis miliacea.

Pharmacological screening of this plant can be divided roughly into the following

major steps:

 Collection and proper identification of the plant.

 Preparation of plant sample.
 Drying and Grinding
 Extraction
 Cold extraction of plant material
 Evaporation of the solvent
 Storage

3.1.1 Collection and Identification

Fimbristylis miliacea was collected from rural region of Manikgonj, Bangladesh on

30th October 2018 and was identified by the expert of National Herbarium Institute,

Mirpur, Dhaka, Bangladesh. (Accession number: DACB-44932).

Figure 3.1: Fimbristylis miliacea

39
3.1.2 Preparation of Plant sample

The collected plant leaves were sun-dried for 10 days. The plant parts were ground

into a coarse powder with the help of a suitable grinder. The powder was stored in

an airtight container and kept in a cool, dark and dry place until analysis

commenced.

3.1.3 Extraction

Crude plant drugs find their way in modern medicine system through continuous

extraction, followed by different isolation techniques and different pharmacological

tests. Chemical constituents from crude plant can be extracted by following two

extraction procedures-

3.1.3.1 Cold Extraction Procedure

In this process powered plant materials are submerged in a suitable solvent or

solvent system in an air tight flat bottom container for several days, with occasional

shaking and stirring. The major portion of plant materials will be dissolved in the

solvent. Solvent is then separated from dispersed materials and evaporated to get

desired extract. In our current study we used cold extraction method.

3.1.3.2 Hot Extraction Procedure

In this process powdered plant materials are successively extracted to exhaustion in

a Soxhlet at elevated temperature with various solvents of increasing polarity.

Individual extract is then filtered through several means. All the extracts are

concentrated with rotary evaporator at low temperature (40-50 °C) and reduced

pressure. Concentrated extract finally obtained is known as crude extract. After cold

or hot extraction, the extract obtained has to be stored in the refrigerator in order to

avoid loss of material.

40
3.1.3.3 Cold extraction of Fimbristylis miliacea
About 500 g of powdered material of the plant was taken separately in a clean, flat-

bottomed glass container and soaked in 2L of 80% Methanol. The container with its

contents was sealed and kept for a period of 21 days accompanying occasional

shaking and stirring. The whole mixture then underwent a coarse filtration by a

piece of clean filter cloth. After that the filtrate was again filtered through cotton to

remove undesirable materials. Then it was filtered through whatman filter paper to

get a fine clear extract solution. The filtrate (methanolic extract) obtained was

evaporated by rotary evaporator until a highly concentrated mixture was found.

Then the concentrate was dried through artificial air to remove the rest solvent and

to get a brownish black color concentrate. That concentrate was designed as crude

methanolic extract of FImbristylis miliacea.

41
 Chapter 4: Materials & Methods
4.1 Chemicals and drugs

All chemicals and reagents used in this study were of analytical grade. Methanol

(Merck, Germany) was used as a solvent during extraction. Glibenclamide (Square

Pharmaceuticals Limited, Bangladesh) was used as standard and Dextrose (Acme

Laboratories Ltd, Bangladesh) were used to produce hyperglycemia. Silymarin

(Square Pharmaceuticals Limited) and Paracetamol (Beximco Pharmaceuticals Ltd.)

were also used in this research.

4.2 Instruments and apparatus

 Rotary Evaporator: Lab Tech-EV311-V

 Sonicator: POWER SONIC 505

 Vacuum pump: Lab Tech-VP30

 Water Heater: Lab Tech-HB-03

 Water Bath: XMTD-204

 Digital Balance: Axis-AGN-220C

 Incubator

 Digital Centrifuge machine

4.3 Experimental Animals

All animal procedures and experimental protocols were approved by the Research

Ethics Committee, Noakhali science and Technology University and were carried

out in accordance with the Guide for the Care and Use of Laboratory Animals

(Dragstedt, 2002). Six-seven weeks old Swiss albino mice of both sexes with mean

body weight 25 ± 5.0 g were procured from Jahangir Nagar University, Savar,

Bangladesh. The animals were housed as 4 in 1 polycarbonate cage in a temperature

42
(23 ± 1) ºC and humidity (55-60%)-controlled room with a 12-h light-dark cycle.

Animals were fed with a commercial rat pellet diet ad libitum during the entire

experimental period. Ethical review board has approved to collect human blood.

4.4 Oral glucose tolerance test

Hypoglycemic effect of leaf extract in mice model was performed according the

method described by Joy and Kuttan (Joy and Kuttan 1999). A group of six mice

was used for each test sample. For Group I (control), mice were given only distilled

water (10 mL/kg of mouse body weight) while Group II was used as standard group

and treated with glibenclamide (10 mg/kg). Group III, IV and V were treated with

leaf extract at doses of 100 mg/kg, 200 mg/kg and 400 mg/kg of mouse body weight

respectively. All animals were fasted overnight before the experiment. The

experiment was started by measuring blood glucose concentration (fasting glucose

concentration) followed by immediate administration (p.o.) of test sample to

respective group and then rested for next 1 h. After this period, dextrose (2 g/kg)

solution was administered to all groups. Blood was collected from mouse tail vein

after 1 h, 2 h and 3 h of glucose administration. Glucose concentration was

measured by Accu-Check electronic glucometer (Roche, Germany). Percentage

reduction in glucose concentration was calculated as (fasting glucose concentration

- glucose concentration at specific time point) x 100/fasting glucose concentration.

4.5 Hepatoprotective activity

4.5.1 Experimental design and animal grouping

Hepatotoxicity was induced by paracetamol induced liver damage model by

Sreedevi et al. (2009). Mices were divided into six groups (n = 5). Group I (normal

control) animals were administered a single dose of water 105 ml/kg, p.o.) daily for

43
7 days and received paracetamol (250 mg/kg, p.o) on day 7. Group II (paracetamol

control) received water (10 ml/kg, p.o.) once daily for 7 days and received

paracetamol (250 mg/kg, p.o) on day 7. Group III received standard drug silymarin

(100 mg/kg, p.o.) once daily for 7 days. Groups IV–VI were administered orally a

dose of 200, 400 and 600 mg/kg of the extract once daily for 7 days respectively.

The Groups III–VI animals were administered simultaneously paracetamol (250

mg/kg, p.o) on day 7 after 2 h of administration of the silymarin and the extract.

After 24 h of treatment, animals were anaesthetized using chloroform inhalation jar.

Than animal were sacrificed and blood was collected through cardiac puncture and

the serum was separated. Liver and kidney were also collected for studies.

Sreedevi, C.D., Latha, P.G., Ancy, P., Suja, S.R., Shyamal, S., Shine, V.J., Sini, S.,

Anuja, G.I., Rajasekharan, S., 2009. Hepatoprotective studies on Sida acuta Burm.

f. J. Ethnopharmacol. 124, 171–175.

4.5.2 Preparation of serum from blood

Blood was drawn by puncturing the cardiac puncture under chloroform anesthesia.

Blood was collected, allowed to clot and serum was separated by centrifugation at

6000 × g for 15 min and analyzed for various biochemical parameters. Serum was

stored in the -800C freezer before they were analyzed.

4.5.3 Determination of key liver function biochemical markers

Liver function biochemical markers such Alkaline phosphatase (ALP), serum

glutamic oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase

(SGPT) and total bilirubin have been evaluated in the serum obtained from the

experimental animals according to the sup-plier’s specifications from the standard

kits. Liver and both kidney weight was also determined in gram.

44
4.6 Statistical analysis
Data were calculated as mean ± SEM values. One-way ANOVA with Dunnett’s test

was done using GraphPad Prism (version 8.0). A probability level of 0.05 or less

was accepted as significant; *p < 0.05, **

p < 0.01, ***
p < 0.001 vs. control.

45
 Chapter 5: Results and discussions
5.1 Hypoglycemic effect

Effect of methanol leaf extract on hyperglycemia is shown in Figure 1 and Table 1.

Leaf extract at all doses showed significant reduction (p < 0.01 for 100 mg/kg, p <

0.001 for 200 mg/kg and p < 0.01 for 400 mg/kg) in blood glucose concentration.

Extract at 400 mg/kg produced time-dependent effect that gradually increased as

time elapsed; similar phenomenon was observed for 100 mg/kg dose. Extract at

doses of 200 mg/kg showed prompt effect at 1 h but effect was slightly reduced

over time. Maximum hypoglycemic effect (36.92%) was observed at 400 mg/kg

extract after 3 h while glibenclamide produced 46.69% reduction.

Effect of plant extract

16 in hyperglyceamic condition
Blood glucose concentration mmol/L)

Control
Glibenclamide
12
FM100
FM200
FM400
8

***
**
4 **
***

0
-1 0 1 2 3
Time (h)
Extract Glucose
administration administration

Figure 5.1: Effect of methanol leaf extract of Fimbristylis miliacea on

hyperglycemic condition. ‘FM’ stands for Fimbristylis miliacea and 100, 200 and
300 indicate doses in mg/kg. A probability level of 0.05 or less was accepted as
significant; *p < 0.05, **p < 0.01, ***p < 0.001 vs. control i.e. distilled water.

46
Table 5.1: Percentage reduction of blood glucose concentration by methanolic
extract of leaf of Fimbristylis miliacea

Reduction of blood glucose concentration

(from fasting glucose concentration)

After 1 h After 2 h After 3 h

Glibenclamide 28.47% 46.69% 46.69%

FM100 0.59% 23.70% 24.87%

FM200 25.70% 23.74% 20.68%

FM400 14.78% 31.18% 36.92%

‘FM’ stands for Fimbristylis miliacea and 100, 200 and 300 indicate doses in
mg/kg.
Different plant extracts of diverse genera and families have been reported to show

hypoglycemic effect due to the presence of various classes of phytoconstituents

such as terpenoid (Uddin, Hasan et al. 2014), alkaloid (Patel and Mishra 2011),

flavonoid (Lü, Chen et al. 2009), glycoside (Chen, Li et al. 2008), etc.

Hypoglycemic effect of plant extracts may be due to stimulation of insulin release

by β-cells or inhibition of glucose absorption from intestine. Leaf extract of

Fimbristylis miliacea is expected to contain important phytoconstituents that offer

potent hypoglycemic effect. Further phytochemical and biological investigations are

required to isolate phytoconstituent responsible for hypoglycemic effect with

mechanism of action.

5.2 Hepatoprotective effect

The estimation of enzymes in the serum is a useful quantitative marker of the extent

and type of hepatocellular damage. The mice were administered with overdose of

paracetamol (250 mg/kg) caused significant liver damage and necrosis of cells as

47
evidenced by the elevated serum hepatic enzymes (ALT, AST and ALP) and

increased level of total bilirubin. The level of enzyme markers ALT, AST and ALP

in normal mice were found to be 42.48 ± 4.81, 186.20 ± 11.79, 9.14 ± 0.96 (U/l)

respectively. Paracetamol intoxication made their elevation to 2.09, 1.79 and 1.32-

fold increment with the values of 88.77 ± 9.42, 332.66 ± 40.13 and 12.03 ± 0.67

U/L respectively compared to the normal control group. This indicates the hepatic

injury and loss of structural integrity. Treatment of animals with MEFM at the doses

of 200, 400 and 600 mg/kg, p.o. or silymarin 100 mg/kg, p.o., significantly

decreased the level of serum marker enzymes (ALT and ALP) and total bilirubin

compared to the Paracetamol intoxicated group. However, there was significant

increase the serum levels of AST in the animals treated with 200, 400 and 600 mg/

kg, respectively compared to the Silymarin. Pre-treatment with extracts significantly

reduced their elevations with the normal values in the range of 17-77, 54-298, and

35-96 U/l for ALT, AST and ALP respectively. Treatments with methanolic

extracts of Fimbristylis miliacea indicate the stabilization of plasma membrane as

well as repair of hepatic tissue damage caused by paracetamol. Similarly, a distorted

pattern for other markers by 5.89-fold increased bilirubin content was observed in

PC mice compared to the NC mice (14.20 vs. 2.41 mg/dl for bilirubin). Right and

left kidney weight was enlarged in Paracetamol intoxicated group compared to

normal group but liver, right and left kidney are significantly regained its normal

size when the animals were treated with MEFM at the doses of 200,400 and 600

mg/kg or silymarin (100 mg/kg), paracetamol control (250 mg/kg) and normal

control (10 ml/kg) mice.

48
Table 5.2: Effect of Fimbristylis miliacea extract in paracetamol induced liver
damage

Treatment Doses ALT AST ALP Total Wt. of Wt. of Wt. of

(U/L) (U/L) (U/L) bilirubin Liver right left

(mg/dl) (gm) kidney kidney

(gm) (gm)

Normal 10 ml/kg, 42.48 186.20 9.14 2.41 0.94 0.13 0.12

control (NC) p.o. ±4.81 ±11.79 ±0.96 ±0.52 ±0.02** ±0.01 ±0.01

Paracetamol 250 88.77 332.66 12.03 14.20 0.80 0.16 0.14

control (PC) mg/kg, p.o ±9.42 ±40.13 ±0.67 ±3.78 ±0.01 ±0.01 ±0.01

100
6.78 129.40 5.12 0.20 0.94 0.13 0.12
PC+Silymarin mg/kg,
±.77** ±1.208 ±1.02** ±0.06 ±0.04 ±0.01* ±0.01
p.o.

34.02 137.02 15.14 0.15 1.11 0.15 0.15

200 mg/kg
±1.94* ±49.69 ±1.23 ±0.06 ±0.13 ±0.14 ±0.01

21.44 207.6 9.65 0.20 1.16 0.16 0.15

PC + MEFM 400 mg/kg
±5.95** ±55.07 ±1.37 ±0.04 ±0.08 ±0.01 ±0.01

12.59 609 8.55 0.51 0.84 0.13 0.12

600 mg/kg
±5.29** ±65.94 ±1.39 ±0.13 ±0.19 ±0.01* ±0.01

MEFM stands for methanol extract of Fimbristylis miliacea and data was presented
as mean ± SEM. ANOVA was employed, followed by Dunnett’s test and significant
differences were represented by *p<0.05, **p<0.01, ***p<0.001 vs paracetamol
control.
It is known that many toxic compounds accumulate in the liver where they are

detoxified (Clarke and Clarke, 1977). Liver transaminases such as AST (aspartate

transaminase) or SGOT (serum glutamic oxaloacetic transaminase), and ALT

(alanine transaminase) or SGPT (serum glutamic pyruvic transaminase) have still

remained the gold standards for the assessment of liver injury, and have been used

as biomarkers of choice for decades (Howell et al., 2014). A study of liver function

tests may therefore prove useful in assessing especially the toxic effects of

49
medicinal plants on the liver. These tests involve mainly the determination of AST

and ALT (Tilkian, 1979) and any marked necrosis of the liver cells leads to a

significant rise of these enzymes in the blood serum. When liver cells are damaged,

these enzymes leak into the bloodstream from liver tissue and produce markedly

elevated serum levels [30]. Both SGOT and SGPT are associated with liver

parenchymal cells. SGPT is found predominantly in the liver with negligible

quantities found in heart, kidneys and skeletal muscles, whereas SGOT is found in

liver, cardiac muscles, skeletal muscles, brain, kidney and red blood cells. Thus

SGPT is a more specific indicator of liver intoxication as levels of SGOT may also

be increased in diseases affecting other organs. On the other hand, serum ALP and

bilirubin levels are related to the functions of hepatic cell. Elevation in level of

serum ALP is due to increased synthesis, in presence of increased biliary pressure.

Our experiment showed that rats which are intoxicated with Paracetamol develop a

significant liver necrosis which was evidenced by increased level of hepatic marker

enzymes (ALT, AST and ALP) and the levels of total bilirubin, whereas levels of

total protein were decreased due to liver injury. The values were well comparable to

the control group. Treatment with F. miliaceae at the concentration of 600 mg/kg

revealed a comparable activity with the reference standard silymarin, a potent

hepatoprotective drug. The result of this study showed that after administration with

methnolic extract of Fimbristylis miliaceae the levels of the serum marker enzymes

(ALT and AST) were restored to normal level, thus indicated that MEFM preserved

the structural integrity of hepatocellular components and protected the liver from

the harmful effect of this hepatotoxin.

50
 Chapter 6: Conclusion
To the best of my knowledge, this is the first report about evaluation of in vivo

hypoglycemic and hepatoprotective of methanol extract of Fimbristylis miliacea

whole plant. These findings suggest that the plant may be a potential source for the

development of new hypoglycemic and hepatoprotective drug which support the

traditional use of this plant. Therefore, the study shows that there is a prospective

future in the use of plants as a source of natural medicine for curing various diseases

due to the presence of medicinally important phyto-constituents in plants.

51
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