Momordica charantia (MC) a traditional medicine used for the treatment of many disorders was fed to streptozotocin (STZ)-induced diabetic male Wistar rats at 2, 5 and 10% of the standard diet. Rats were then observed for 12 weeks before sacrificed. Immediately, tissues from liver, kidney and pancreas were taken for histopathological examination. Serum samples were analyzed to evaluate liver, kidney and pancreatic functions. In addition, blood samples were collected to analyze hematology parameters. The results of the present study indicate that oral doses of MC at 5% and 10% of the daily diet resulted in alleviation of the pancreatic, hepatic and renal dysfunction induced by diabetes. The improved pancreatic, hepatic and renal functions were judged by histopathological, hematological and serobiochemical parameters. In conclusion, the MC fruits may be used as an antidiabetic herbal medicine.
Diabetes Mellitus (DM) is a major metabolic disorder characterized by chronic hyperglycemia as a result of a relative or absolute lack of insulin or its actions [
It is estimated that in the year 2013 more than 382 million people worldwide had DM and 592 million people will subsequently have the disease in 2035 [
It has been shown that the oral hypoglycemic treatment has characteristic profiles of side effects [
Momordica charantia (MC) (
The hypoglycemic effects of MC have been investigated [
Fresh green whole fruits of Momordica charantia (MC) were purchased from local markets in Al-Ahssa, Kingdom of Saudi Arabia. The fruits were sliced and then oven dried at 60˚C temperature for 24 hours. The dried fruit slices were powdered then added to the powdered feed at 2, 5 and 10%.
Streptozotocin (STZ) single dose (85 mg/kg) (Sigma, S0130-USA) freshly dissolved in 0.9% normal saline solution was injected intra-peritoneal to induce diabetes in all rats [
Fifty male Wistar albino rats weighing (150 - 200 g) were housed in hygienic fiberglass cages. Animals were fedon balanced commercial pellets. All rats were given two weeks adaptation period with free access to food and water before starting experimental procedures.
Fifty rats were allotted at random to five groups, 10 rats each:
Group (0), negative control, given untreated diet and water ad-Libitum.
Group (1), positive control given STZ only.
Group (2), given STZ then fed with MC at 2% of the diet.
Group (3), given STZ then fed with MC at 5% of the diet.
Group (4), given STZ then fed with MC at 10% of the diet.
At the end of the experiment (12 weeks) rats from each group were humanly sacrificed, blood samples were collected for serobiochemical and hematological analysis, gross lesions were recorded and tissue specimens from liver, kidney and pancreas were collected and fixed in 10% neutral buffered formalin for histopathological studies.
Blood samples were collected for biochemical parameters. Serum was separated by centrifugation of the clotted blood and stored at −20˚C till used. Samples were then analyzed for the activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP), as well as the concentration of cholesterol, total protein, albumin, globulin, total bilirubin, blood urea nitrogen (BUN), uric acid and creatinine, using Abaxis Vetscan VS2―America analyzer.
Blood samples were collected in test tubes containing ETDA (Ethylene diamine tetra acetic acid) for determination of hemoglobin concentration (HGB), total erythrocyte count (TEC), packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC) and total white blood cells (WBCs) using Abaxis Vetscan HM5―America analyzer.
Tissue specimens from liver, kidney and pancreas were trimmed and put in the vacuum infiltrating tissue processing machine (Tissue-Tik VIP 5Jr. Japan) and embedded in paraffin wax by SLEE MPS/C machine, Germany. Specimens waxed blocks were sectioned to 5 μm by LEICA RM 2235 microtome, Germany and then stained with Hematoxylin and Eosin (H&E) for histopathological examination [
Data were statistically evaluated with SPSS 7.5 software. All results were expressed as mean ± SD [
During the experimental time, group (1) and group (2) showed polyuria, polydipsia, polyphagia and weakness. Moreover, the mortality was approximately 50% in group (1) and 20% in group (2).
On the other hand, group (3) and group (4) showed gradual alleviation of polyuria and polyphagia without weakness and death in both groups. However, rats of group (0), negative control remained healthy throughout the days of experiment. They showed normal urine excretion, normal water intake and food consumption.
Renal hypertrophy and liver congestion were common findings in group (1) and group (2).
1) Hepatic changes
Liver tissues of the control untreated rats (Group 0) appear normal without any histological alterations, (
2) Renal changes
In group (1), massive vacuolation was seen in cortical and medullar renal tubules,
3) Pancreatic changes
The pancreatic tissues of the control untreated rats (Group 0), showed normal islets cells, (
pancreatic tissue were observed in the islets of Langerhans. Rats in group (1) showed necrosis, degeneration and disappearance of islets cells represented by empty spaces and few numbers of remaining cells (
Rats in group (1) showed a significant decrease in the values of total erythrocyte count (TEC), hemoglobin concentration (HGB) and packed cell volume (PCV). Whereas, all treated rats in groups, (2, 3 and 4) showed a significant gradual, dose-dependent increase in the values of Total Erythrocyte Count (TEC), hemoglobin concentration (HGB) and packed cell volume (PCV). It seems that total erythrocyte count (TEC), hemoglobin concentration (HGB) and packed cell volume (PCV) in rats of groups 3 & 4 were significantly higher than that of group 2, (
Haematological parameters | Group (0) | Group (1) | Group (2) | Group (3) | Group (4) |
---|---|---|---|---|---|
TLC [103/µl] | 12.09 ± 0.52a | 11.49 ± 0.81a | 11.62 ± 0.05a | 11.03 ± 0.26a | 11.81 ± 0.14a |
LYM [103/µl] | 8.32 ± 0.36a | 8.58 ± 0.07a | 8.71 ± 0.14a | 8.68 ± 1.45a | 8.54 ± 1.83a |
MON [103/µl] | 0.35 ± 0.17a | 0.37 ± 0.65a | 0.36 ± 0.23a | 0.37 ± 0.61a | 0.36 ± 0.34a |
NEU [103/µl] | 2.79 ± 0.79a | 2.21 ± 0.38a | 2.46 ± 0.26a | 2.40 ± 0.61a | 2.80 ± a0.80a |
TEC [106/µl] | 8.93 ± 0.25a | 4.74 ± 0.22b | 6.17 ± 0.87bc | 7.73 ± 0.43bd | 8.88 ± 0.14a |
HGB [g/dl] | 14.34 ± 0.43a | 9.43 ± 0.40b | 11.25 ± 0.43bc | 12.65 ± 0.61ac | 15.40 ± 0.36ad |
PCV | 49.3 ± 1.35a | 26.84 ± 1.39b | 42.59 ± 1.48cd | 46.91 ± 4.75ad | 50.33 ± 0.54a |
MCV [fl] | 53.25 ± 1.18a | 56.50 ± 2.53ac | 69.50 ± 0.65b | 59.50 ± 1.69dc | 56.60 ± 0.51ac |
MCH [pg] | 16.09 ± 0.42a | 19.83 ± 0.56bd | 18.23 ± 1.52bd | 16.38 ± 0.64a | 17.34 ± 0.38a |
MCHC [g/dl] | 30.38 ± 0.74a | 35.80 ± 0.47b | 26.15 ± 0.69c | 26.75 ± 0.48c | 30.56 ± 0.43a |
PLT [103/µl] | 521.88 ± 66.18a | 577.00 ± 9.06a | 587.75 ± 27.86a | 547.00 ± 42.01a | 547.80 ± 47.45a |
Values are mean ± standard error. Different letters between group means values are significant (p ≤ 0.05).
As shown in
In the present study, polyuria, polydipsia, polyphagia, weakness and death were the common clinical signs observed in group 1 (given only STZ) and group 2 (given STZ then treated with 2% MC). However, alleviation of these clinical signs was observed in group 3 (given STZ then treated with 5% MC) and group 4
Biochemical parameters | Group (0) | Group (1) | Group (2) | Group (3) | Group (4) |
---|---|---|---|---|---|
ALT [IU/L] | 39 ± 1.96a | 272 ± 10.75b | 180.25 ± 6.87c | 88.17 ± 5.65d | 86.00 ± 8.70d |
AST [IU/L] | 67.54 ± 3.73a | 119.75 ± 7.36b | 96.63 ± 9.76c | 89.17 ± 6.14cd | 80.56 ± 2.17ad |
ALP [IU/L] | 131.11 ± 6.16a | 1003.13 ± 25.83b | 803.75 ± 23.20bd | 776.50 ± 30.30cd | 715.00 ± 23.26cd |
T. Bilirubin [mg/dl] | 0.30 ± 0.00a | 0.33 ± 0.03a | 0.30 ± 0.00a | 0.32 ± 0.02a | 0.30 ± 0.00a |
Cholesterol [mg/dl] | 60.78 ± 3.41a | 127.75 ± 8.87b | 101.25 ± 6.34c | 72.71 ± 5.38a | 67.17 ± 4.33a |
Glucose [g/dl] | 117.67 ± 3.21a | 582.00 ± 18.47b | 478.00 ± 15.88c | 399.83 ± 10.51d | 291.00 ± 13.19e |
T. Protein [g/dl] | 7.77 ± 0.15a | 7.55 ± 0.49a | 7.18 ± 0.09a | 7.50 ± 0.23a | 7.79 ± 0.23a |
Albumin [g/dl] | 4.49 ± 0.07a | 3.50 ± 0.13ac | 4.05 ± 0.17ac | 3.57 ± 0.15ac | 3.63 ± 0.40ac |
Globulin [g/dl] | 3.30 ± 0.14a | 4.05 ± 0.60ac | 3.08 ± 0.24ad | 3.92 ± 0.21ad | 4.19 ± 0.26ac |
BUN [mg/dl] | 17.67 ± 0.71a | 37.14 ± 3.89b | 32.25 ± 0.85c | 31.81 ± 2.60c | 28.50 ± 3.97d |
Creatinine [mg/dl] | 0.40 ± 0.04a | 0.85 ± 0.26bc | 0.63 ± 0.06ac | 0.58 ± 0.03ac | 0.51 ± 0.06a |
Uric acid [mg/dl] | 5.16 ± 0.26a | 15.60 ± 0.85b | 13.55 ± 0.52bd | 12.20 ± 0.18cd | 9.30 ± 0.73ce |
Values are mean ± standard error. Different letters between group means values are significant (p ≤ 0.05).
(given STZ then treated with 10% MC). This finding may indicate that MC has promising effects in prevention as well as delay in progression of diabetic complications in rats. These results agree with that obtained by [
Anemia is the most common blood disorder in diabetes mellitus [
Rats in group 1 (given only STZ) showed an increase in concentration of blood glucose (hyperglycemia). Whereas, all treated rats with 2%, 5% and 10% MC, groups 2, 3 and 4 respectively, showed remarkable recovery represented by gradual reduction of blood glucose. This finding is consistent with the results obtain by [
In the present study, all treated rats with 2%, 5% and 10% MC, groups 2, 3 and 4 respectively, showed remarkable decrease in the levels of ALT, AST, ALP, cholesterol BUN, creatinine and uric acid. The reduced levels of these intracellular enzymes after administration of MC to rats may be due to the alleviation of cell plasma membrane damage produced by diabetes. These findings were parallel with the results obtained by [
Rats in group 1 (given only STZ) showed hepatocytomegaly and coagulative necrosis. These findings agree with the results obtained by [
In diabetic rats, hyperglycemiais the main cause of nephropathy, indicated by glomerular hypertrophy [
In addition, [
The islets of Langerhans in group 1 (given only STZ) showed necrosis, degeneration and disappearance indicated by empty spaces and few numbers of remaining cells. These results are parallel with the results observed by [
In conclusion, Momordica charantia (MC) fruits given at 5% and 10% of the daily standard diet for 3 months seem to possess beneficial effects on diabetic rats through alleviation of tissue injury and improvement of hematological and serobiochemical parameters. However rats given 2% MC showed lower degree of improvement in hematological and biochemical parameters compared to the groups given 5% and 10% MC.
One of the limitations of such studies is the use of STZ at (85 mg/kg) to induce diabetes. This dose may cause harmful effects in organs other than the pancreas and may affect the serobiochemical findings specially glucose level. Therefore, the use of MC in animals with naturally occurring diabetes is preferred.
The researchers are grateful to Deanship of Scientific Research, King Faisal University (KFU), Saudi Arabia for funding this project.
Moqbel, M.S., Al- Hizab, F.A. and Barakat, S.M. (2017) Clinicopathological Study on the Effects of Momordica charantia on Streptozotocin-Induced Diabetic Wistar Rats. Open Journal of Veterinary Medicine, 7, 49-62. https://doi.org/10.4236/ojvm.2017.75006