In vivo hypoglycemic effect of methanolic fruit extract of Momordica

charantia L
Nkambo W1, *Anyama NG, Onegi B

Department of Pharmacy, School of Health Sciences, College of Health Sciences, Makerere University, P.O.
Box 7072, Kampala, Uganda

Abstract
Background: Momordica charantia L. is a medicinal plant commonly used in the management of diabetes mellitus.
Objectives: We investigated the blood glucose lowering effect of the methanolic fruit extract of the Ugandan variety of M.
charantia L. in alloxan-induced diabetic albino rats.
Methods: 500g of M. charantia powder were macerated in methanol and the extract administered to two groups of alloxan-
induced diabetic rats. The first group received 125mg/kg, the second 375mg/kg and a third group 7mg/ kg of metformin.
A fourth group received 1ml normal saline. Fasting blood glucose (FBG) levels were measured at 0.5,1,2,3,5,8 and 12 hours
and compared using one-way ANOVA.
Results: There was an initial rise in FBG for 1 hour after administration of extracts followed by steep reductions. Significant
reduction in FBG occurred at 2 hours for 125mg/kg of extract (-3.2%, 31325.9 to 30325.0mg/dL, p = 0.049), 375mg/
kg of extract (-3.9%, 35619.7 to 34220.3mg/dL, p = 0.001), and metformin (-2.6%, 34421.7 to 33521.1mg/dL, p =
0.003) when compared to normal saline. The maximum percentage reduction in FBG by both extracts occurred between 3
and 12 hours post dose.
Conclusions: The methanolic fruit extract of M. charantia exhibits dose dependent hypoglycaemic activity in vivo.
Key words: Momordica charantia, methanolic extract, Diabetes Mellitus, hypoglycaemic effect, in vivo
African Health Sciences 2013; 13(4): 933 - 939 http://dx.doi.org/10.4314/ahs.v13i4.11

Introduction
Diabetes Mellitus (DM) is a leading cause of illness Major risk factors are similar to those of other
and death in developed countries and is epidemic in regions of the world such as urbanization, obesity,
many developing and newly industrialized countries. physical inactivity, or others that are not quite mutable
Its macrovascular and microvascular complications such as increasing age and ethnicity. Most African
are debilitating. The prevalence of diabetes in the countries still face a number of problems related to
world at all ages was estimated to be 2.8% in 2000, the management and treatment of the disease, such
and it is expected to approximate 4.4% in the year as critical shortage of diabetes medicine, the rising
2030.The estimated global number of people of all cost of drugs and treatment, competition for
ages and sex with diabetes in 2000 was 171 million. resources by HIV/AIDs, tuberculosis, and malaria.
This is projected to increase to 366 million by 2030, The general lack of equipment to diagnose the disease
with about 4 million deaths every year attributed to has hampered efforts to manage and control diabetes
its complications1,2. mellitus3,2 . In Uganda, there is a new surge of non-
The estimated number of people with communicable diseases, among them diabetes and
diabetes in sub-Saharan Africa was 10.8 million in this is partly due to changing lifestyle. Many urbanites
2006, and this could rise to 18.7 million by 2025. neither exercise nor do physical work. The number
*Corresponding author: of people with diabetes is now thought to have
Anyama G Norbert passed a million, with 560,000 registered patients and
Department of Pharmacy it is thought an equal number unknowingly have
School of Health Sciences undiagnosed disease4,5.
College of Health Sciences, Makerere University Anti-diabetic treatments or interventions are
P.O. Box 7072 grouped into three major categories; diet and exercise
Kampala, Uganda. which form part of first line treatment of diabetes,
Tel: +256-312213113 insulin and oral hypoglycaemic agents. However, the
Email: nanyama@chs.mak.ac.ug latter are often expensive and inaccessible to many

African Health Sciences Vol 13 Issue 4 December 2013 933

low-income generating individuals in Uganda, given machine and weighed 138.6g.The powder was put
their high cost and the sometimes long distance that in a clean empty bottle and methanol added until it
has to be travelled to the hospitals and health facilities covered the powder, with vigorous shaking to mix
that avail them. Also, these drugs are not without the content. Methanol was added to make 2 litres.
side effects and yet the treatment is life-long, due to The bottle was then corked and kept for 3 days
the chronic nature of disease. Because of this, some with occasional shaking to facilitate extraction of the
patients use affordable and cost effective alternative active component from the powder. A 2 litre
therapy for management of diabetes in the form measuring cylinder and funnel and round bottomed
of traditional medicines, which are both locally flask were cleaned and dried. The cotton was placed
available and cheap2, 4. in the neck of the funnel and placed on top of the
Several herbal remedies used in the management of cylinder. The macerating mixture was poured into
diabetes have been reported to possess hypoglycemic the funnel to filter off the large size marc. The process
effects6-11. Among these is Momordica charantia L Fam. was repeated on the filtrate using Whartman filter
Cucurbitaceae (African cucumber, bitter gourd, bitter paper and the filtrate collected in a round bottomed
melon) a medicinal plant used traditionally as an flask. The dry extract was obtained using a rotary
antidiabetic, an emetic, a laxative, a tonic, and to treat evaporator. The percentage yield was 17 percent.
anaemia, arthritis, colds, fever, gout, infertility, kidney The extract was stored in a vial in a cupboard.
stones, peptic ulcers, stomach ache and intestinal Methanol was used because it is a polar
helminthes12 . It is also used as an antimalarial, together solvent and so hopefully would extract active
with related species, and as an abortifacient13-15. Some principles, which otherwise would have been
pharmacological and safety studies of this herb have extracted using water. Also, it is easier to evaporate
been carried out16,17. In addition to hypoglycaemic compared to water.
activity, M.charantia has been shown to have
antioxidant18-20, anti-tumour21-25, neuroprotective26, Preparation of the extract and metformin
anti-inflammatory27-29 and antimicrobial activity30,31. 5g of extract were suspended in 25ml of normal
It has a resistance modifying effect for saline solution to form a 200mg/ml suspension. One
aminoglycosides against methicillin-resistant tablet of metformin (500mg) obtained from a
Staphylococcus aureus32. The plant is a source of urease pharmacy was powdered and the powder dissolved
for urea determination33. Wan et al34 have also found in 50ml of normal saline to form a 10mg/ml
that M. charantia peroxidase can be used for suspension.
biotransfor mation of piceatannol into
antihyperglycaemic oligomeric stilbenes. Animal preparation
This study aimed at investigating the effects Twenty four male albino rats weighing between 150
of the Ugandan variety of M. charantia L. methanolic and 180g were chosen in order to provide uniform
fruit extract on blood glucose levels in alloxan- results and minimize error that occurs due to
induced diabetic rats. variation in species, sex and weight. The animals were
obtained from the School of Veterinary Medicine
Methods and Animal Resources, Makerere University and
Plant collection and extraction habituated at the Department of Pharmacology,
M. charantia ripe fruits were obtained from College of Health Sciences, in cages for 3 days under
Kabanyolo farm at the beginning of the dry season normal laboratory conditions of; temperature,
(December 2011 to February 2012).The herbarium humidity and light (12 hours day, 12 hours night).They
specimen was prepared and verified at the were fed on standard animal feed and water ad
Department of Botany, Makerere University. Studies libitum.
have shown that hybridization occurs between
cultivated and wild varieties and that there is transfer Induction of Diabetes Mellitus in animals
of genetic material between species35,36. The fresh The animals were made to fast for 18 hours receiving
fruits were washed with tap water to remove dust only water. They were weighed and the Fasting Blood
and other foreign material. They were then air dried Glucose (FBG) of each animal measured by bleeding
in the laboratory. The dry fruits were blended into a the diethyl ether anesthetized animal on the tail and
powder form using a mortar and pestle. The recording the glucose level using a glucometer. 1.5g
powder was weighed using a digital weighing of alloxan powder, purchased from BDH

934 African Health Sciences Vol 13 Issue 4 December 2013

laboratories, was dissolved in 25ml of 0.9% normal Statistical analysis
saline to form a 60mg/ml solution. This was put in The means of fasting blood glucose levels for the
a vial, autoclaved at 1210C for 3 hours and cooled. test and control groups were compared at different
Specific volumes of the solution were taken off and times by one-way analysis of variance (ANOVA)
injected into the tail veins of diethyl ethyl anesthetized using SPSS 11 software. A p value <0.05 was
animals such that each animal got 65mg/kg body considered statistically significant.
weight37. The animals were then monitored for 5
days and elevation of FBG confirmed after 18 hours. Ethical considerations
Only animals with FBG above 200mg/dL were This study was approved by the Institutional Review
used in the study. Committee of the School of Medicine, College of
Health Sciences, Makerere University. All experiments
Administration of test substances were conducted in accordance with internationally
Twenty four animals were randomly assigned to 4 accepted principles for animal use and care.
groups of 6 each namely I, II, III and IV, and fasted
for 18 hours. Using a syringe and endogastric tube, Results
suspensions of the extract were administered by There was an initial increase in the FBG when the
gavage to restrained animals such that group IV extract was administered, which lasted the first 1 hour.
received 1ml normal saline, group III 7mg/kg body The rise was greater with 125mg/kg (13.0%) than
weight of metformin suspension, and groups I and 375mg/kg (8.9%) of the extract. Thereafter, there
II 125mg/kg and 375mg/kg body weight of M. was a significant reduction in FBG at 2 hours for
charantia respectively. 125mg/kg of extract (-3.2%, 31325.9 to
30325.0mg/dL, p = 0.049), 375mg/kg of extract
Measurement of Fasting Blood Glucose (-3.9%, 35619.7 to 34220.3mg/dL, p = 0.001),
Blood drops were obtained by piercing the tip of and metformin (-2.6%, 34421.7 to 33521.1mg/
diethyl ether anesthetized tails of the rats and FBG dL, p = 0.003) when compared to normal saline
measured using a glucometer. The fasting blood (figure 1). The maximum percentage reduction in
glucose levels were measured at 0, 0.5,1,2,3,5,8 and FBG by both extracts occurred between 3 and 12
12 hours after administration of the substances. hours post dose (table 1).

Figure 1: Mean blood glucose (mg/dL) after administration of methanolic fruit extract of Momordica
charantia and metformin in alloxan-induced diabetic rats (n=6)
SE = standard error of mean

African Health Sciences Vol 13 Issue 4 December 2013 935

Table 1: Percentage glycaemic change after administration of methanolic fruit extract of Momordica
charantia L. in alloxan-induced diabetic rats (n=6)

Treatment Percent glycaemic change*

Time (Hours)
0 0.5 1 2 3 5 8 12
Normal saline 1ml 0 -2.7 -2.8 0.4 -4.1 4.3 -4.5 -4.3
Metformin 7mg/kg 0 -3.5 -2.8 -2.6 -2.4 -3.4 -2.8 -3.3
M. charantia extract 125mg/kg 0 4.3 8.3 -3.2 -5.3 -6.9 -6.2 -10.0
M. charantia extract 375mg/kg 0 4.6 4.1 -3.9 -6.7 -4.4 -5.9 -13.6
*Negative values indicate reductions

The subsequent FBG reductions 2 hours after in fasting blood glucose levels 42 . However, a
administration of extract remained significant up to systematic review of four Randomized Controlled
12 hours for 375mg/kg, while that of 125mg/kg Trials of M. charantia for type 2 DM by Ooi et al43
of extract was significant after the third hour. The showed no difference with placebo, metformin or
percentage reductions for both concentrations of glibenclamide indicating the need for further clinical
the extract were greater than those of metformin studies, standardization and quality control of
between 8 and 12 hours (14% and 10% as compared preparations.
to 3% respectively). The effect on FBG of 375mg/ In this study, the anti-hyperglycaemic effects
kg of the methanolic extract was comparable to that of metformin (7mg/kg) and 375mg/kg extract
of metformin (p > 0.05 between 2 and 12 hours). were more or less similar. While metformin lowers
FBG concentrations by decreasing hepatic
Discussion gluconeogenesis and increasing insulin-stimulated
The antidiabetic effect of M. charantia was glucose uptake by skeletal muscle and adipose tissues,
investigated and the results show that at 2 hours, M. charantia appears to act by repairing damaged
both concentrations of the methanolic fruit extract Beta-cells, increasing insulin secretion, enhancing
exhibited declines in blood glucose, with 375mg/ insulin sensitivity in peripheral tissue by promoting
kg of extract having a greater effect than 125mg/ glucose uptake, inhibition of hepatic
kg of extract. The onset of glucose lowering was gluconeogenesis, decreasing glucose absorption by
not as rapid as with metformin, yet the trajectory inhibiting glucosidase and disaccharidases in the
appeared superior. Both the extract and metformin intestine, and enhancing the activity of AMP-activated
lowered blood glucose levels without inducing protein kinase44 . Indeed some of the constituents
hypoglycaemia. The initial rise in blood glucose could of the extract like oleanolic acid 3-O-glucuronide
be attributed to the carbohydrate content of the plant and momordin exert their anti-hyperglycemic effect
or as a result of a physiological phenomenon38. This by inhibiting glucose transport at the brush border
was not observed in mice administered normal of the small intestine. The aqueous extract of the
saline. This initial rise in FBG seems to offset the unripe fruit of M. charantia has been shown to
early anti-hyperglycaemic effect of the crude extract. partially stimulate insulin release from isolated Beta-
Kolawole et al39 showed that the methanolic cells of the pancreas in rats, while the fruit juice
fruit extract of M. charantia decreased blood glucose significantly increased the number of Beta-cells6. M.
in both normal and diabetic animals, comparable to charantia has also been reported to inhibit 11Beta-
10mg/kg of chlorpropramide in doses of 400 to hydroxysteroid dehydrogenase, a potential anti-
600mg/kg. Mamun 40 also found a significant diabetes target45.
decrease in blood glucose and increase in serum Major active principles in M. charantia are
insulin when powdered fruits of the plant were sterols, triterpenes, glycosides notably momordin Ic,
administered to diabetic rats, while Rathnaker et al 41 charantin, goyaglycosides, momordicosides and
have demonstrated the hypoglycaemic effect of a other cucurbitane glycosides, goyasaponins, the
polyherbal product containing M. charantia. In alkaloid momordicin, phenolic compounds, tannins,
another study, a different species, Momordica cymbalaria flavonoids, carotenoids and bioactive proteins like
was found to produce a time-dependent decrease polypeptide p and alpha-momorcharin12,44,46-50. The

936 African Health Sciences Vol 13 Issue 4 December 2013

oleanane-glycoside momordin Ic and cucurbitane- 4. Hjelm K, Atwine F. Health-care seeking behavior
type triterpenoid glycosides especially charantin and among persons with diabetes in Uganda: an
polypeptide p have been shown to have interview study. BMC International Health &
hypoglycaemic activity44,51-54. While Harazika et al55 Human Rights 2011; 11:11
have demonstrated that momordicilin a triterpene, 5. Mutebi E, Nakwagala FN, Nambuya A, Otim
is a potent inhibitor of glycogen synthase kinase-3, M. Undiagnosed diabetes mellitus and impaired
an enzyme involved in glucose homeostasis and glucose tolerance among hypertensive patients
potential target for anti-diabetic compounds. in Mulago Hospital, Kampala, Uganda. African
These findings provide further evidence for Journal of Diabetes Medicine 2012; 20(1): 20-23
hypoglycemic activity of M. charantia similar to that 6. Grover JK, Yadav S, Vats V 2002. Medicinal
seen in other members of the Cucurbitaceae Family. plants of India with anti-diabetic potential.
Journal of Ethnopharmacology 81: 81100.
Limitations 7. Macedo M, Ferreira AR. Plantas
It is noteworthy that initial blood glucose levels were hipoglicemiantes utilizadas por comunidades
slightly different for extract, metformin and saline tradicioanais na Bacia do Alto Paraguai e Vale o
groups at baseline. This was difficult to control. Guapore, Mato Grosso-Brasil. Rev Bras Farmacogn
However, we determined the rate and extent of 2004; 14(supl. 01): 45-47
decrease in blood glucose, which was greater for 8. Djomeni Dzeufiet PD, Tedong L, Asongalem
the metformin and extract groups compared to EA, Dimo T, Sokeng SD, Kamtchouing P.
normal saline. Hypoglycaemic effect of methylene chloride/
methanol root extract of Ceiba pentandra in
normal and diabetic rats. Indian Journal of
Conclusion
Pharmacology 2006; 38: 194197
The study revealed that the methanolic fruit extract
9. Nwaegerue E, Ifeoma NN, Ezeala CC,
of M. charantia exhibited anti-hyperglycaemic effects
Unekwe PC. Glucose lowering effect of leaf
comparable to those of metformin, in appropriate
extract of Viscum album in normal in normal
doses, in alloxan-induced diabetic rats, but the initial
and diabetic rats. J Res Med Sci 2007; 12(5): 235-
effect appears to be offset by the carbohydrate
340
content of the extract. The anti-hyperglycaemic
10. Rajasekhar MD, Ramesh Babu K, Vinay K,
activity increased with an increase in dose of extract.
Sampath MR, Sameena SK, Apparao C.
Antihyperglycemic and antioxidant activities of
Acknowledgement active fraction from the aqueous extract of
We would like to thank Aloysius Lubega for his Momordica cymbalaria fruits in Streptozotocin
assistance in the laboratory studies. induced diabetic rats. Phcog Res 2009; 1: 352-8
11. Verissimo LF, Bacchi AD, Zaminelli T, Henrique
Funding O. de Paula G, Moreira EG. Herbs of interest
This research project was funded by the to the Brazilian Federal Government: female
Pharmaceutical Society of Uganda reproductive and developmental toxicity studies.
Rev Bras Famacogn 2011; 21(6): 1163-1171
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