Journal of Applied Pharmaceutical Science Vol. 5 (04), pp. 007-012, April, 2015
Available online at http://www.japsonline.com
DOI: 10.7324/JAPS.2015.50402
ISSN 2231-3354
Evaluation of the cytotoxic activity of extracts from medicinal plants
used for the treatment of malaria in Kagera and Lindi regions,
Tanzania
Ramadhani S. O. Nondo1*, Mainen J. Moshi1, Paul Erasto2, Denis Zofou3,4, Abdel J. Njouendou4, Samuel Wanji4, Moses
N. Ngemenya3, Abdul W Kidukuli1, Pax J. Masimba1, Vincent P.K. Titanji3
1
Department of Biological and Pre–Clinical Studies, Institute of Traditional Medicine, Muhimbili University of Health and Allied Sciences, Dar es Salaam,
Tanzania. 2National Institute for Medical Research, Dar es Salaam, Tanzania. 3Biotechnology Unit, University of Buea, Buea, South West Region,
Cameroon. 4Research Foundation in Tropical Diseases and Environment, Buea, South West Region-Cameroon.
ARTICLE INFO
ABSTRACT
Article history:
Received on: 06/02/2015
Revised on: 18/02/2015
Accepted on: 09/03/2015
Available online: 27/04/2015
A number of medicinal plants used for treatment of malaria in Tanzania have been documented, but information
on their safety and efficacy is still based on traditional knowledge accumulated over years and not on pre-clinical
and clinical evaluation. The present study aimed to assess the cytotoxic activity of extracts of selected plant
species used for treatment of malaria in Tanzania. Ethanol extracts were evaluated for cytoxicity by using MTT
assay on LLC-MK2 cells and by brine shrimp lethality assay. Forty five (93.75%) out of 48 crude extracts
assessed using LLC-MK2 cells were non-cytotoxic while three extracts (6.25%) were cytotoxic with CC50 <30
µg/mL (cut-off point). In the brine shrimp assay 30 (65.2%) out of 46 extracts tested were non-toxic while 16
extracts (34.8%) were toxic (LC50 <100 µg/mL). Antiaris toxicaria stem bark extract was the most cytotoxic to
mammalian cells. This study demonstrates that, most of the antimalarial plants tested were non-toxic. These
observations corroborate with traditional healers’ claims that the herbal medicines used in their areas are safe.
However, further studies using different toxicity models are suggested to further confirm their claims.
Key words:
Toxicity, medicinal plants,
malaria, LLC-MK2 cells,
brine shrimps assay,
Tanzania
INTRODUCTION
The use of plants as source of medicines for treatment
of infectious and non-infectious diseases is an old human
tradition (Petrovska, 2012), and the practice is now increasing
due to increased global health challenges (WHO, 2002). Malaria
is one of diseases treated by herbal medicines originating from
different plant parts such as roots, stem bark, leaves, flowers and
fruits. It is an old life threatening parasitic disease caused by
parasites of the genus Plasmodium. The parasites infect and
destroy red blood cells, leading to fever, severe anaemia,
cerebral malaria and death may occur if the patient is not treated
properly and on time (Fidock et al., 2004; NIAID, 2007).
* Corresponding Author
Ramadhani SO Nondo, Department of Biological and Pre–Clinical
Studies, Institute of Traditional Medicine, Muhimbili University of
Health and Allied Sciences, Dar es Salaam, Tanzania.
Email: nondo75@yahoo.com
Exploration of the accumulated indigenous knowledge on the
treatment of malaria using medicinal plants enabled the isolation of
two important and currently used antimalarial drugs; artemisinin
from Artemisia annua and quinine from the bark of Cinchona spp
(Wells, 2011). Despite plants being a rich source of useful chemical
compounds of various structures and with different
pharmacological properties on biological systems (Butler, 2004;
Moshi et al., 2009), some of them may be toxic to humans. For
example, some of the toxicities associated with the use of medicinal
plants include allergic reactions, irritation of the gastrointestinal
tract, destruction of red blood cells, and damage of body organs
such as the heart and kidney and carcinogenicity (Westendorf,
1999; IARC, 2012). Several medicinal plants have previously been
reported to be toxic. Some of the examples include Symphytum
officinale L. used for wound healing which contains hepatotoxic
pyrrolizidine alkaloids and Valerian officinalis used as a sedative
for treatment of insomnia and anxiety which causes hepatitis
© 2015 Ramadhani SO Nondo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License -NonCommercialShareAlikeUnported License (http://creativecommons.org/licenses/by-nc-sa/3.0/).
08
Nondo et al. / Journal of Applied Pharmaceutical Science 5 (04); 2015: 007-012
(Abdualmjid and Segi, 2013). Aristolochia spp contain aristolochic
acid I and II that cause renal failure (Debelle et al., 2008); Drimia
sanguinea and Bowiea volubilis which are traditionally used for
headache, oedema, infertility and bladder problems contain
cardiotoxic bufadienolides (Van der Bijl Jr. and Van der Bijl Sen.,
2012). Although the use of herbal medicines is controlled in many
countries, information about their efficacy and safety is based on
traditional knowledge transmitted through generations over years
and not on pre-clinical and clinical evaluation (Chalut et al.,
1999). Tanzania shares the same experience of having a number of
traditional healers who use traditional medicines for treatment of
different diseases and is endowed with over 12,000 plant species,
of which at least 10% have medicinal values (Mahunnah et al.,
2012). Furthermore, Tanzania is among the six African countries
with many reported cases of malaria (WHO, 2012) and because of
the long history of the disease, the practice of using medicinal
plants to treat malaria is very common (Mahunnah, 1987; Gessler
et al., 1995 Kinung’hi et al., 2010). Although several antimalarial
medicinal plants have been documented in Tanzania, their safety
has not been well studied. Therefore in this study the toxicity of
crude extracts of medicinal plants used for the treatment of malaria
in Kagera and Lindi regions, Tanzania, were assessed using the
LLC- MK2 monkey kidney epithelial cell line and the brine
shrimp larvae (Artemia salina L.).
MATERIALS AND METHODS
Materials
Monkey kidney epithelial cells, LLC-MK2 (ATCC®,
USA) were obtained from American Type Culture Collections
(USA) and ethanol (Carlo erba®) was purchased from Techno Net
Scientific (Dar es Salaam, Tanzania). Foetal Bovine Serum (FBS,
BioWhittaker®, Verviers, Belgium), RPMI-1640 medium (Sigma),
MTT
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl
tetrazolium
bromide (Sigma), sodium bicarbonate (sigma), dimethylsulfoxide
(Sigma), cell culture flasks (Corning, NY, USA), 96 well cell
culture plates (Costar®, Corning, NY, USA), Centrifuge tubes
(Corning, NY, USA), Syringe adapted filters 0.22µm (Corning,
NY, USA), Trypsin-EDTA (Sigma) were all purchased from
Sigma (Steinheim, Germany). Plant materials were collected from
Kagera (November, 2012) and Lindi (July, 2012) regions,
Tanzania. Identification of the plants was done by Mr. Haji. O.
Selemani, a Botanist at the Department of Botany, University of
Dar es Salaam, and the voucher specimens are deposited at the
University of Dar es Salaam and at Muhimbili University of
Health and Allied Sciences Herbaria, Tanzania.
Extraction of crude extracts
The powdered plant materials were macerated in 80%
ethanol at room temperature for 24h and then filtered through
cotton wool. The solid plant materials were macerated again in the
same solvent for another 24h and the extracts obtained from the
first and the second extractions were pooled and concentrated
under vacuo using a Heldolph® rotary evaporator (Heldolph
instruments GmbH, Schwabach, Germany) to obtain viscous
extracts which were further dried using a freeze drier (Edwards
High Vacuum International, Crawley Sussex, England). The dry
extracts were stored at -20°C until use.
Preparation of stock solutions
Stock solutions were prepared by dissolving 4 mg of
crude extracts in 100 µL dimethyl sulfoxide and then diluted with
RPMI-1640 cell culture medium to make 400 µg/mL. All solutions
were sterilized by passing through 0.22 µm syringe-adapted filters
and stored at -20°C until use.
Determination of cytotoxic activity on LLC-MK2 cells
Cytotoxicity of the crude extracts was evaluated on LLCMK2 monkey kidney epithelial cells. Cells were grown in RPMI1640 culture medium with L-glutamine and 25 mM HEPES
(Steinheim, Germany). The medium was supplemented with 2
mg/mL NaHCO3 (sigma), 10 µg/mL hypoxanthine (Sigma), 11.1
mM glucose (sigma), 10% FBS (BioWhittaker®, Verviers,
Belgium) and 5µg/mL gentamicin. The cells were incubated at 5%
O2, 5% CO2, and 90% N2 in humidified incubator (SHEL LAB™,
Sheldon Mfg Inc, OR, USA) at 37°C until confluent before used
for cytotoxicity assay. Trypsinated cells were distributed in 96
well plates at 10,000 cells in 100 µL per well and incubated for 48
h to allow them to attach before adding the extract. After 48 h the
medium was removed completely from each well, and 100 µL of
fresh culture medium was then added. Thereafter 100 µL of crude
extracts (400 µg/mL) were added in row H and then serially
diluted to row B to give concentrations ranging from 200 – 3.125
µg/mL. Cells in row A served as controls without drug (100%
growth). The cells with or without extracts were incubated at 37°C
for 72 h before determining their viability. Each concentration
level was tested in triplicate.
MTT Assay
Cell viability was determined using MTT assay (Niles et
al., 2008; 2009). After 72 h of incubation, the culture medium in
each well with or without extract was removed completely from
the assay plates and replaced by 100 µL of fresh culture medium.
Then 10µL of 5 mg/mL Thiazolyl Blue Tetrazolium Bromide,
MTT (Sigma) were added into each well to achieve a final
concentration of 0.45mg/mL before incubated for 3 h at 37°C.
After 3 h, the culture medium with MTT was carefully removed
followed by addition of 100µ L dimethylsulfoxide to dissolve
formazan crystals and then incubated for 1 h before recording the
optical density (Emax-Molecular Devices Corporation, California,
USA) at 595 nm.
Data analysis
The percentage viability and percentage mortality were
calculated from the OD values using Microsoft Excel 2010. The
mean results of the percentage mortality were plotted against the
logarithms of concentrations using the Fig P computer program
Ver 4.189/07 (Biosoft Inc, USA). Regression equations obtained
Nondo et al. / Journal of Applied Pharmaceutical Science 5 (04); 2015: 007-012
from the graphs were used to calculate the fifty percent cytotoxic
concentration (CC50), which is the concentration killing fifty
percent of the cells. An extract with CC50>30 µg/mL is considered
non-toxic (Fadeyi et al., 2013).
Brine shrimp toxicity assay
The brine shrimp lethality assay is a non-specific toxicity
assay that is used in natural products research to detect the
presence of pharmacologically active chemical constituents. It uses
Artemia salina L. (Artemiidae) larvae (Meyer et al., 1982).
Solutions of plant extracts were made in dimethylsulfoxide. The
brine shrimp toxicity assay was conducted and data analyzed as
previously reported (Nondo et al., 2011). An LC50 (concentration
killing fifty percent of the brine shrimp larvae) value greater than
100 μg/mL is considered to represent a non-toxic compound or
extract (Moshi et al., 2010). Each extract was tested in duplicate
and the concentrations of dimethylsulfoxide were restricted to a
maximum of 0.6% in the final volume.
RESULTS AND DISCUSSION
Forty eight extracts from 38 medicinal plants distributed
into 19 different plant families were evaluated for cytotoxic
activity on mammalian cells (LLC-MK2 cells) as presented in
Table 1. The results revealed that 45 (93.75%) out of 48 extracts
tested were non-cytotoxic and exhibited CC50 values above the
cut-off point which is 30 µg/mL. Of these, 33 extracts (73.3%)
from 24 plant species were found to have CC50 values above 200
µg/mL, the highest concentration tested. Only three (6.25%) out 48
extracts were found to be cytotoxic with CC50 < 30 µg/mL (cut-off
point). The extracts were from Aspilia natalensis aerial parts
(18.57 ± 1.04 µg/mL), Antiaris toxicaria leaves (12.51 ± 0.65
µg/mL) and Antiaris toxicaria stem bark (1.44 ± 0.48 µg/mL)
(Table 1).
The brine shrimp toxicity assay showed that thirty
extracts (65.2%) out of the 46 extracts tested had LC50 values
greater than 100 µg/mL; the cut-off point. Among these, 8 extracts
had LC50 values greater than 1000 µg/mL, while the remaining had
LC50 values between 100 and 800 µg/mL. Only Sixteen extracts
(34.8%) showed LC50 <100 µg/mL, and therefore classified as
toxic. Maesa lanceolata leaf extract was the most toxic with LC50
= 1.55 µg/mL, followed by the extracts from Dalbergia
malangensis leaves (16.47 µg/mL), Aspilia natalensis aerial parts
(34.93 µg/mL), Desmodium salicifolium stem (36.87 µg/mL), and
Dalbergia malangensis stem extract (47.59 µg/mL) (Table 2). The
high toxicity of M. lanceolata leaf extract on brine shrimp larvae
may be due to the effect of saponins. Previous studies revealed that
the leaves of M. lanceolata are rich in triterpenoidal saponins and
these compounds were reported to have high molluscicidal and
hemolytic activities (Sindambiwe et al., 1998; Apers et al., 2001).
According to the American National Cancer Institute (NCI), a
crude plant extract is considered to be cytotoxic if its CC50 value
on mammalian cells is <30 µg/mL (Fadeyi et al., 2013). On the
other hand the cut-off point to consider a crude plant extract non-
09
toxic in the brine shrimp toxicity assay is LC50>100 µg/mL (Moshi
et al., 2010). Based on the results obtained in the two bioassays,
three extracts were found to be toxic against LLC-MK2 cells and
16 extracts were found to be toxic on brine shrimp larvae. Of these
only one extract from A. natalensis aerial parts was found to be
toxic in both assays, which may be an indicator of consensus for
cytotoxicity.
The extract from A. toxicaria stem bark was ranked the
most toxic on the mammalian cells (LLC-MK2 cells) but it was
ranked as exceptionally non-toxic using the brine shrimp toxicity
assay (with LC50 >1,000 µg/mL). On the other hand M. lanceolata
leaf extract was ranked as the most toxic on brine shrimp assay
with LC50 = 1.55 µg/mL but ranked as non-toxic on LLC-MK2
cells test (Table 1 and 2). These observations suggest that the two
models used in this study complement each other for the detection
of toxic compounds that may be attributed to different mechanisms
of toxicity; although the brine shrimp bioassay was found to be
more sensitive in detecting toxic extracts than LLC-MK2 cells.
The difference may be explained partly by the non-specificity of
the brine shrimp assay in detecting toxic compounds (Meyer et al.,
1982) and the differences in the criteria set to define a toxic
substance, although in some studies brine shrimp assay has been
reported to demonstrate some correlation with cell line results for
detecting cytotoxic compounds/extracts (Meyer et al., 1982;
Carballo et al., 2002).
The cytotoxicity of A. natalensis aerial parts extract was
predicted by both assays; but it exhibited higher toxicity to
mammalian cells than to brine shrimp larvae. The cytotoxic
activity of A. natalensis (CC50 = 18.57 ± 1.04 µg/mL) on LLCMK2 cells was comparable to that of the standard cytotoxic drug
used in this study (Imatinib, Gleevec) which had CC50 of 18.61
µg/mL. A previous study revealed that an infusion and paste
prepared from leaves of A. natalensis are used topically in South
Africa to treat skin diseases (Mabona et al., 2013), but information
regarding its toxicity was limited. Information from the traditional
healers who reported these plants indicated that the decoctions of
A. natalensis leaves and A. toxicaria leaves and stem bark are used
orally for malaria associated with high fever (‘‘Malaria kali’’).
They, however, emphasized that the decoction of A. natalensis
should be consumed in small quantity because if taken in large
quantities it causes stomach pain. These results may support the
safety concern raised by traditional healers regarding oral
administration of extracts from this plant.
Antiaris toxicaria is a known poisonous plant used in
arrow poisoning associated with the presence of a number of
cardiac glycosides which are inhibitors of Na+/K+ -ATPase pump
(Kopp et al., 1992; Shi et al., 2010). In addition, the cardiac
glycosides and coumarins isolated from A. toxicaria were reported
to have cytotoxic activity on various cancer cell lines (Dai et al.,
2009; Liu et al., 2013; Shi et al., 2014). In this study we found that
ethanolic extracts of the leaves and stem bark of A. toxicaria were
very toxic to non-cancer cells (LLC-MK2). However, these results
do not support the questionnaire-based toxicity information
collected from traditional healers.
010
Nondo et al. / Journal of Applied Pharmaceutical Science 5 (04); 2015: 007-012
Table 1: Cytotoxic activity of crude extracts on LLC-MK2 cells (CC50 ± SD in µg/mL).
CC50 ± SD (µg/mL)
80% EtOH crude extract
Acanthaceae
Acanthus pubescens (Oliv.) Vatke
Amatoju
>200
Funtumia africana (Benth) Staff
Mwezamaino/omwelamaino
>200
Apocynaceae
Funtumia africana (Benth) Staff
Mwezamaino/omwelamaino
>200
Holarrhena pubescens (Huch-Ham)
Nalupande
>200
Holarrhena pubescens (Huch-Ham)
Nalupande
>200b
Burseraceae
Canarium schweinfurthii Engl.
Omubafu wa kike/muubani wa kike
>200
Celastraceae
Salacia lovetii N. Halle & B. Mathew
Omzindabikaka
>200
Aspilia mosambecensis (Oliv.) Wild
Eshurwa rusharila/Esisa
>200
Compositae
Aspilia natalensis (Sond) Wild
Kanyamoisa
18.57 ± 1.04
Guizotia scabra (Vis.) Chiov
Echihongosheija
>200
Vernonia glabra (Steetz) Vatke
Msangusangu
100.75 ± 16.69
Convolvulaceae
Ipomoea rubens choisy
Kataba
>200
Bridelia micrantha (Hochst.) Bail
Omushamako
156.80 ± 0.44
Euphorbiaceae
Phyllanthus nummulariifolius Poir
Karungi
>200
Phyllanthus nummulariifolius Poir
Karungi
>200a
Cassia singueana
Mlewelewe
>200b
Cassia singueana
Mlewelewe
>200
Dalbergia malangensis E.P. Sousa
Omugorora
107.29 ± 11.04
Dalbergia malangensis E.P. Sousa
Omugorora
>200
Fabaceae
Desmodium salicifolium (Poir) DC
Batengeliange/Omukongoranwa
>200
Erythrina sacleuxii Hua
Mlindimila/mnungunungu
>200
Erythrina schliebenii Harms
Mlindimila
>200
Erythrina schliebenii Harms
Mlindimila
>200a
Erythrina schliebenii Harms
Mlindimila
>200
Macrotyloma axillare (E. Mey) Verdc
Akaihabukuru
>200
Leonotis nepaetifolia (L.) R. Br
Ekitatelante
130.04 ± 0.23
Labiatae
Leonotis nepaetifolia (L.) R. Br
Ekitatelante
137.80 ± 2.29
Leonotis nepaetifolia (L.) R. Br
Ekitatelante
124.13 ± 11.86
Logamiaceae
Anthocleista grandiflora Gilg
Omubagaigana/mbagaigana
>200
Dissotis brazzae Cogn
Bulitulo
134.47 ± 2.7
Dissotis melleri Hook. f.
Ekituntun/Etuntun
83.33 ± 3.31
Melastomataceae
Dissotis rotundifolia (Sm) Triana
Obwehehe/Obwee
125.90 ± 1.86
Melastomatrum capitatum (Vahl) A.& R. Fern)
Katuntun/akatuntun
>200
Melianthaceae
Bersama abyssinica
Omujalya
>200
Antiaris toxicaria (Pers) Lesch
Omujuju
1.44 ± 0.48
Moraceae
Antiaris toxicaria (Pers) Lesch
Omujuju
12. 51 ± 0.65
Pycnanthus angolensis (Welw.) Warb
Omunonoba
136.32 ± 3.09
Myristicaceae
Pycnanthus angolensis (Welw.) Warb
Omunonoba
>200
Pycnanthus angolensis (Welw.) Warb
Omunonoba
>200
Myrsinaceae
Maesa lanceolata Forsk
Omuzilanyama/omuhanga
141.86 ± 2.02
Myrtaceae
Syzygium cordatum Krause
Omugege
>200
Rosaceae
Eriobotrya japonica (Thunb.) Lindl
Musharazi/Omusharazi
>200
Pentas bussei (K. Krause)
Rusharila kibira
>200
Hallea rubrostipulata (K. Schum) J.F.Leny
Mchunguchungu
141.69 ± 0.61
Hallea rubrostipulata (K. Schum) J.F.Leny
Mchunguchungu
>200
Rubiaceae
Oxyanthus speciosus DC
Omwanikibira
>200
Rhytignia obscura Robyns
Omulokola/lulokola
>200
Rutaceae
Teclea amaniensis
>200
Standard drug: Gleevec (Imatinib)
18.61 ± 1.30
a= aqueous extract, b= methanol extract. All other extracts are extracted by 80% ethanol. CC50= cytotoxic concentration fifty percent (mean ± SD, n =3). R = root,
S = stem, SB = stem bark, L = leaves, AP = aerial parts (stem + leaves), F = fruits, FL = flowers, WP = whole plant
Plant family
Plant species
Vernacular name
During our ethnobotanical survey, traditional healers
reported that decoctions of leaves and stem bark were non-toxic
when taken orally for treatment of malaria. This information from
the reporting traditional healers is supported by animal studies.
Kang et al., (2008) reported that aqueous and ethanolic leaf
extracts of A. toxicaria were not toxic to mice even at high doses
when given orally. But toxicity was observed when these extracts
were administered by intra-peritoneal route. This may suggest that
the bioavailability of the cardiac glycosides present in leaves and
stem bark is low when given orally compared to when given
through other routes. Apart from the toxicity evaluation reported in
Plant
part
R
SB
L
R
R
SB
L
AP
AP
WP
L
L
SB
WP
WP
R
R
L
S
AP
SB
SB
SB
R
AP
FL
L
AP
SB
AP
AP
AP
AP
SB
SB
L
F
SB
L
L
SB
L
AP
R
SB
L
L
R
this study, the plant extracts reported in this study were previously
evaluated for in vitro antimalarial activity against P. falciparum
Dd2 strains. At a single concentration of 100 µg/mL, ethanolic
extracts from A. toxicaria stem bark, M. lanceolata leaves, A.
natalensis and D. salicifolium aerial parts inhibited the growth of
malaria parasites in vitro (Nondo et al., 2015). Since the LLCMK2 cells are normal mammalian cells, toxicity against these cells
most likely predicts lack of selectivity and thus it will be toxic to
mammalian cells, and therefore the traditional healers and patients
should be informed on the risk of toxicity that might arise
following use of extracts from these plants.
Nondo et al. / Journal of Applied Pharmaceutical Science 5 (04); 2015: 007-012
011
Table 2: Brine shrimp toxicity results.
Plant species
Part
LC50 (µg/mL)
95% CI (µg/mL)
Acanthus pubescens (Oliv.) Vatke
R
140.94
113.20 – 175.47
Athocleista grandiflora Gilg
SB
> 1,000
Antiaris toxicaria (Pers) Lesch
L
154.24
112.18 – 212.08
Antiaris toxicaria (Pers) Lesch
SB
>1,000
Aspilia mosambecensis (Oliv.) Wild
AP
122.17
93.98 – 158.82
Aspilia natalensis (Sond)
AP
34.93
25.68 – 47.50
Bersama abyssinica
SB
729.14
433.75 – 1,225.68
Bersama abyssinica
R
184.35
126.88 – 267.86
Bridelia micrantha (Hochst.) Bail
SB
>1,000
Canarium schweinfurthii Engl.
SB
273.51
203.66 – 367.32
Cassia singueana
R
> 1,000
332. 36
241.19 – 458.0
Cassia singueana
Rb
Dalbergia malangensis E.P. Sousa
L
16.47
10.78 – 25.17
Dalbergia malangensis E.P. Sousa
Stem
47.59
39.89 – 56.77
Desmodium salicifolium (Poir) DC
AP
36.87
28.96 – 46.94
Dissotis brazzae Cogn
AP
244.39
187.69 – 318.17
Dissotis melleri Hook. f.
AP
116.75
89.26 – 152.71
Dissotis rotundifolia (Sm) Triana
AP
>1,000
Eriobotrya japonica (Thunb.) Lindl
L
>1,000
Erythrina schliebenii Harms
SB
729.14
433.75 – 1,225.68
Erythrina schliebenii Harms
R
93.26
74.97 – 116.02
Funtumia africana (Benth) Staff
SB
223.06
175.64 – 283.29
Funtumia africana (Benth) Staff
L
348.56
223.29 – 544.10
Guizotia scabra (Vis.) Chiov
WP
60.14
43.49 – 83.17
Hallea rubrostipulata (K. Schum) J.F.Leny
SB
125.45
99.01 – 158.95
Hallea rubrostipulata (K. Schum) J.F.Leny
R
>1,000
Holarrhena pubescens (Huch-Ham)
Ra
291.58
204.76 – 415.21
Holarrhena pubescens (Huch-Ham)
R
63.16
54.77 – 72.82
Holarrhena pubescens (Huch-Ham)
Rb
135.24
111.40 – 164.19
Ipomoea rubens choisy
AP
97.01
73.05 – 128.83
Leonotis nepaetifolia (L.) R. Br
AP
128.74
90.92 – 182.30
Leonotis nepaetifolia (L.) R. Br
FL
91.75
68.62 – 122.67
Macrotyloma axillare (E. Mey) Verdc
AP
123.91
89.60 – 171.37
Maesa lanceolata Forsk
L
1.55
0.59 – 4.08
Melastomatrum capitatum (Vahl) A.& R. Fern)
AP
390.17
267.79 – 568.48
Oxyanthus speciosus DC
L
229.48
152.28 – 345.83
Pentas bussei (K. Krause)
AP
729.14
433.75 – 1,225.68
Phyllanthus nummulariifolius Poir
WP
86.14
67.83 – 109.40
87.39
68.02 – 110.03
Phyllanthus nummulariifolius Poir
WPa
Pycnanthus angolensis (Welw.) Warb
F
81.01
52.74 – 124.43
Pycnanthus angolensis (Welw.) Warb
L
78.55
68.19 – 90.49
Pycnanthus angolensis (Welw.) Warb
SB
93.58
74.98 – 116.79
Rhytignia obscura Robyns
L
489.33
296.20 – 608.37
Syzygium cordatum Krause
SB
99.93
80.33 – 124.31
Vernonia glabra (Steetz) Vatke
L
>1,000
LC50 = lethal concentration fifty, CI = confidence interval, a= aqueous extract, b= methanol extract. All other extracts were extracted by 80% ethanol. R = root,
S = stem, SB = stem bark, L = leaves, AP = aerial parts (stem + leaves), F = fruits, FL = flowers, WP = whole plant.
CONCLUSION
Most of the antimalarial medicinal plants tested were
non-toxic, and hence support the traditional healers’ claims who
believe that the herbal medicines they use are safe. However,
further studies using different toxicity models are suggested to
confirm their claims. Only the extracts of A. natalensis and
A. toxicaria were categorized as toxic to mammalian cells. The
evidence of A. natalensis toxicity obtained in this study supports
the cautionary note that was given by the collaborating traditional
healers.
The authors also wish to acknowledge traditional healers
(Mr. Mohamed Ngalanga, Mr. Didas Ngemera, Mr. Dominic
Mushwahili, Mr. Buchadi Tibikunda and Mr. Papianus
Rwechungura) and the Botanist (Mr. Haji.O. Selemani) for their
assistance in collecting plants used in this study. Extraction of the
plant extracts and brine shrimp toxicity assay were done at the
Institute of Traditional Medicine in Tanzania; whereas toxicity
study on LLC-MK2 cells was done at the Research Foundation in
Tropical Diseases and Environment, and at the University of Buea
in Cameroon.
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How to cite this article:
Nondo R.S.O., Moshi M. J., Erasto P., Zofou D., Njouendou A. J.,
Wanji S., Ngemenya M.N., Kidukuli A.W., Masimba P.J., Titanji
V.P.K. Evaluation of the cytotoxic activity of extracts from
medicinal plants used for the treatment of malaria in Kagera and
Lindi regions, Tanzania. J App Pharm Sci, 2015; 5 (04): 007-012.