Professional Documents
Culture Documents
Pakistan, Philippines, Sri Lanka, Thailand and Vietnam. No published results are
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
while extract at 400 mg/kg showed 36.92% reduction after same time span.
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
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
iii
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
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’
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
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
starting with the 1526 Grete Herball. John Gerard wrote his famous ‘The Herball or
2
Culpeper published his The English Physician Enlarged. Many new plant medicines
Europe, translating and copying classical texts and maintaining herb gardens.
the Islamic Golden Age, scholars translated many classical Greek texts including
Abulcasis (936–1013) of Cordoba wrote The Book of Simples, and Ibn al-Baitar
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
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
medicinal, it is implied that the said plant is useful as a drug or therapeutic agent or
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
health),
3. Tannins (used for gastro-intestinal problem like diarrhea, ulcer and for
inflammation and protect affected tissues against further injury and cause
7. Vitamins and minerals (Fruits and vegetables are the sources of vitamins and
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
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
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
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.
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
hippocastanum
relaxant
paniculata
analgesic, anti-
6
inflammatory
proteolytic
camphora
relaxant
gout
7
Digoxin Cardiotonic Digitalis purpurea
antihistamine
inhibitor
disease
astringent
agent
8
Lanatosides A, B, C Cardiotonic Digitalis lanata
respiratory stimulant
(topical)
paniculata
acid
detoxicant
Inhibitor
9
relaxant
agent
antipyretic
tranquillizer
tranquillizer
tranquillizer
10
inhibitor Canadensis
traquillizer
Phytotherapy, the treatment of disease by the use of plants, was the beginning 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
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
enumerated as -
1st stage: Crude drugs were employed, prepared in the roughest manner,
2nd stage: Crude drugs were converted into more active and manageable
3rd stage: The pure active principles separated from the crude drug were
4th stage: Attempt to synthesize the active drug in the laboratory and indeed
occur (IUCN, 2003). The number of medicinal plants included in the materia
and more than five hundred of such medicinal plants has so far been enlisted as
12
Figure 1.2: Major medicinal plants production zones in Bangladesh
(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
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
ayurvedic companies also reported about some sporadic farming of medicinal plants
in different areas of Bangladesh. One of the ayurvedic company reported that some
Government formed a cell for medicinal plant in the ‘Ministry of Environment and
a) Promoting implantation,
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
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
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
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
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
pure morphine from natural opium in 1803. Pure quinine was isolated from
important plant derived drugs of modern medicine rapidly followed and many
useful drugs have since been discovered and introduced into modern medicine.
Theasinensis (1819), quinine from Chcinchona spp. (1820) and colchicines from
Colchicum autumnale (1820) constitutes some example of such early drugs (Ghani
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
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
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
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
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
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
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
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).
Plants, the molecular architect still offers a great potentiality for drug search. Plants
with pharmacologic activity. Digitalis, opiates and the cinchona alkaloids (quinine
and quinidine) came into modern medicine by this route. Curare was obtained from
Rawolfiaserpentina was used for a variety of illnesses; only in recent years its
practice report numerous medicinal plants, which are still not investigated. These
18
methods can be applied to isolate the responsible agent. Bioactivity guided
ones.
procedures, all fractions are tested, and active fractions are further
possess the claimed properties and exert the desired therapeutic effects. A
programs for drug discovery. Emergence of newer disease also leading the scientists
order to substantiate the above assertion about their validity these medicinal plants
constitute the major constituents of indigenous medicines. Thus, the quality and
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
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
further increased their importance in the field of medicine, for example in the
these it is possible to modify the activity or regulate the properties of the key
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
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,
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
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
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
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.
cure.
21
Cadaba fruticosa L. Capparaceae Shrub Root decoction is
administered during
helminthiasis
internal haemorrhages
dysentery
eczema
Rheumatism
22
Chloroxylon swietenia Rutaceae Tree Leaves used in Rheumatism
skin troubles
troubles
rheumatoid arthritis
asthma,diarrhea
in rheumatism
nervous debility.
piles
23
Cajanus cajan L. Fabaceae Shrub Leaf extract is administered
abdominal pains
cough
diarrhea
asthma
bone fracture
dysentery
24
Quisqualis indica L. Combretaceae Shrub Seeds used for treating
helmenthiasis
administered for
haemorrhoids
snake bite
troubles
and other synthetic chemistry techniques which has been funding by pharmaceutical
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.
pharmacological and clinical tests are pre-requisites for developing drugs from
medicinal plants. Modern drug development from plant source is carried out
fractions.
26
There have no pharmacological studies have been performed to assess the
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
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
rhizomes and stolons are clothed with scales which usually disintegrate leaving
fibrous remains.
2.1.1 Morphology
28
Spikelets of Cyperaceae the individual flowers are subtended by a single
scale.
Fibrous roots.
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.
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
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
30
Scirpodendron, Scirpoides, Scirpus, Scleria, Sphaerocyperus, Stenophyllus,
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
The Plant List for the genus Fimbristylis taken from the version 1.1 (Published on
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
- Scirpus miliaceus L.
35
- Scirpus parviflorus Willd. ex Kunth
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
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
The spikelets flower and then develop tiny fruits, which are brown achenes.
2.2.5 Distribution
Lanka, Thailand and Vietnam. It has also been introduced into Ecuador,
2.2.6 Habitat
Paddy fields, common in moist to swampy places or in shallow water, along ditches,
37
Fimbristylis falcata (Vahl) Kunth; Rhizome (juice) in dysentery.
rheumatism.
treat fever.
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:
30th October 2018 and was identified by the expert of National Herbarium Institute,
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
tests. Chemical constituents from crude plant can be extracted by following two
extraction procedures-
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
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
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
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
41
Chapter 4: Materials & Methods
4.1 Chemicals and drugs
All chemicals and reagents used in this study were of analytical grade. Methanol
Incubator
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,
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.
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
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
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.
mg/kg, p.o) on day 7 after 2 h of administration of the silymarin and the extract.
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.
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
(SGPT) and total bilirubin have been evaluated in the serum obtained from the
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
45
Chapter 5: Results and discussions
5.1 Hypoglycemic effect
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.
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
Control
Glibenclamide
12
FM100
FM200
FM400
8
***
**
4 **
***
0
-1 0 1 2 3
Time (h)
Extract Glucose
administration administration
46
Table 5.1: Percentage reduction of blood glucose concentration by methanolic
extract of leaf of Fimbristylis miliacea
‘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
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.
mechanism of action.
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
increase the serum levels of AST in the animals treated with 200, 400 and 600 mg/
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
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
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
48
Table 5.2: Effect of Fimbristylis miliacea extract in paracetamol induced liver
damage
(gm) (gm)
control (NC) p.o. ±4.81 ±11.79 ±0.96 ±0.52 ±0.02** ±0.01 ±0.01
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.
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
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
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
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
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
50
Chapter 6: Conclusion
To the best of my knowledge, this is the first report about evaluation of in vivo
whole plant. These findings suggest that the plant may be a potential source for 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
51
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