Source: http://biomedpharmajournal.org/vol10no3/evaluation-of-antidiabetic-activity-of-aqueous-and-ethanolic-extracts-of-leaves-of-chloroxylon-swietenia-in-streptozotocin-stz-induced-diabetes-in-albino-rats/
Timestamp: 2019-04-20 04:40:33+00:00

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Kadali S. L .D. V. R. M, Das M. C, Vijayaraghavan R, Kumar M. V. Evaluation of Antidiabetic Activity of Aqueous and Ethanolic Extracts of Leaves of Chloroxylon Swietenia in Streptozotocin (Stz) Induced Diabetes in Albino Rats. Biomed Pharmacol J 2017;10(3).
1Department of Research, Saveetha University, Chennai.
2Department of Pharmacology, NRI Medical College, Guntur, AP.
3Department of Research, Saveetha University, Chennai.
4Department of Pharmaceutical Chemistry, NGSM Institute of Pharmaceutical Science, Deralakatte, Mangalore.
Toevaluate antidiabetic activity of Chloroxylon swietenia in STZ induced diabetes in albino rats Forty two albino rats were randomly divided into seven groups (n=6). Diabetes was induced by intraperitoneal injection of streptozotocin (60mg/Kg). Distilled water, Tween 80, glibenclamide, Chloroxylon swieteniaaqueous extract(CSAE), ethanolic extracts (CSEE) of 200 and 400mg/kg were given orally for 14 days to the normal control, diabetic control, standard group and test groups respectively. Glucose, TC (total cholesterol), TG (triglyceride), HDL (High density lipoprotein), AST (aspartate aminotransferase), ALT (alanine aminotransferase), ALP (alkaline phosphatase), creatinine, direct bilirubin and indirect bilirubinlevels were estimated. ANOVA followed by Student-Newman-Keuls test was used to analyze the data. CSAE of 200mg/kg showed a significantreduction in glucose, ALT, TB and ALP levels in diabetic rats.CSAE of 400mg/kg showed a significant decrease in glucose, AST, ALT, TB, DB and ALP levels in diabetic rats.CSEE of 200mg/kg showed a significant decrease in glucose, ALT, TB,ALPandcreatininelevels in diabetic rats.CSEE of 400mg/kg showed a significant decrease in glucose, AST, ALT, TB, DB and ALP levels in diabetic rats. Both extracts show antidiabetic activity in STZ induced diabetes.
An ethnobotanicalstudy was carried on the medicinal plants often used for the management of diabetes in Warangal district, Andhra Pradesh by traditional healers.Chloroxylon swieteniais the one of the plant used by the traditional healers for diabetes.7,8Even though medicinal plants are widely used, the effective treatment of the disease has not been verified with scientific standards. Only a few plants usedfor diabetesin traditional medicineare scientifically audited in vivo.9Chloroxylon swietenia belongs to the Rutaceae family. Common name – satinwood, Telugu name – billu, bildu,billedu, Tamil name- porasu orvaaimaram.Chloroxylon swietenia has been reported to have anti-inflammatory activity,10 mosquitocidal activity,11-13 antioxidant activity,14 analgesic activity,14 anthelmintic activity15,antimicrobial activity.15-17Antidiabetic activity was reported with this plant, but with different parts of stem, bark and whole plant.18,19Invitro antidiabetic activity was reported with the leaf extract of Chloroxylon swietenia in our previous report.20Based on the claims and available evidence, it was thought worthwhile to investigate Chloroxylon swietenia for diabetes in animal models.
The leaves of Chloroxylon swietenia were collected locally and authenticated by Dr. Shiva Kumari, Department of Botany, Andhra Loyola College.After shade-dried (Temp<40°C.), plant material was grounded into a moderately coarse powder. The aqueous extract was made by maceration and the ethanolic extract was made by using soxhlet apparatus. The extract was allowed to dry. The dried extract was weighed. The % yield of each plant extract was calculated. The % of yield obtained was 8.96 and 9.16% for alcoholic and aqueous extracts respectively. Both the extracts were preserved in the refrigerator till further use.
Both sexes of albino rats weighing 250-300g were used. Rats werefedwith a standard pellet diet and water ad libitum. Animals were kept in a controlled environment (12 h/12 h light/night) and temperature (27±2°C).Before starting the experiment, rats were allowed to acclimatize to the laboratory conditions. All the animal experiments were approved by the institutional animal ethics committee(36/IAEC/NRIMC/2013-14) in accordance with the guidelines of the Committee for the Purpose of Control and Supervision of Experimentation on Animals.
STZ was freshly prepared by dissolving in citrate buffer (0.01M, PH-4.5) and kept on ice prior to practice. The overnight fasted rats were made diabetes with a single intraperitoneal injection of STZ (60 mg/kg). After 4hrs STZ administration 5% glucose was administered orally in drinking water for a day to overcome the early hypoglycemic phase.Rats were allowed to stabilize for three days.On the third day (72hrs)blood samples were drawn to estimate the blood glucose concentration to confirm the development of diabetes.Rats with plasma glucose above 250 mg/dL were considered as diabetic and used in the study.Both the test extracts and standard drug treatment were given orally for 14days. Blood was collected by the retro-orbitalpuncture under light ether anesthesia on 1, 7 & 14th day of treatment schedule for biochemical estimations.
Rats were randomly allocated into 7groups (n=6) (Table-1).
The data were expressed as mean ± standard error (SE). The Significance of differences among the groups were assessed by using ANOVA, followed by Student-Newman-Keuls test. p<0.05 (5%) were considered as significant.
The effect of extracts on glucose level is illustrated in Graph-1. Statistical analysis at ‘0’ (zero) dayby One-way ANOVA revealed that there was no significant (P>0.05) difference among the groups. Further, statistical analysis on the 7thday of medication showed a significant (P<0.05) difference among the groups. There was a significant elevation of blood glucose level in diabetic control as compared to normal control rats.Student-Newman-Keulstest revealed that glibenclamide, CSAE1, CSAE2, CSEE1 and CSEE2treated groups shows asignificant reduction in blood glucose level as compared to the diabetic control. Similarly, statistical analysis at 14thday showed that there was significant (P<0.05) difference among the groups. Student-Newman-Keuls test revealed that glibenclamide, CSAE1, CSAE2, CSEE1 and CSEE2treated groups shows asignificant reduction in blood glucose level as compared to the diabetic control. Further analysis by ANOVA followed by Student-Newman-Keuls test revealed that glibenclamide, CSAE1, CSEE1 and CSEE2treated groups shows no significant difference between the groups when test groups are compared with standard (glibenclamide) control. This result indicates that test groups produced the effect almost equal to the standard group.
Graph 1: The effect of glibenclamide, CSAE and CSEE on plasma glucose (FBS) in diabetic rats.Mean + SE (n = 6). a Statistically significant from control group. b Statistically significant from STZ group.
The changes in the AST and ALT levels of all the groups are illustrated in Graph-4. Statistical analysis by One-way ANOVA revealed that there was asignificant difference among the groups [p<0.05].There was a significant elevation of AST and ALT level in diabetic control as compared to normal control rats. Glibenclamide, CSAE1, CSAE2, CSEE1 and CSEE2treated groups show asignificant reduction in ALT level as compared to the diabetic control. CSAE1 and CSEE1reduced the AST level when compared to the diabetic control, butthe reduction is not statistically significant (p>0.05). Both the extracts of 400mg/kg dose shows significant(p<0.05) reduction in the AST levels.
Graph 2: The effect of CSAE and CAEEon TC, TG, HDL, LDL and VLDL in diabetic rats.Mean + SE (n = 6).a Statistically significant from control group. b Statistically significant from STZ group.
The changes in the total bilirubin and direct bilirubin levels of all the groups are illustrated in Graph-5.There was a significant elevation of total bilirubin and direct bilirubin levels in diabetic control as compared to normal control rats.CSAE1, CSAE2, CSEE1 and CSEE2 treated groups shows asignificant reduction in the total bilirubin level as compared to the diabetic control. CSEE of 400mg/kg dose shows the significant reduction inthe direct bilirubin level when compared to the diabetic control.
Graph 3: The effect of glibenclamide, CSAE and CSEE on AI, CRI and % protection in diabetic rats.Mean + SE (n = 6). a Statistically significant from control group. b Statistically significant from STZ group.*Statistically significant from glibenclamide group.
ALP and creatinine levels are significantly elevated in diabetic controls as compared to the normal control (Graph-4 and 5). Both extracts of all the doses significantly reduced the elevated ALP levels. CSEE of 200mg/kg showed asignificant decrease in the creatinine levels.
Graph 4: The effect of glibenclamide, CSAE and CSEE on AST, ALT and ALP in diabetic rats.Mean + SE (n = 6). a Statistically significant from control group. b Statistically significant from STZ group.
Graph 5: The effect of glibenclamide, CASE and CSEE on total bilirubin (TB), direct bilirubin (DB) and creatinine (Cr) in diabetic rats.Mean + SE (n = 6). a Statistically significant from control group.b Statistically significant from STZ group.
There was a significant rise in TC, TG, VLDL and LDL levels in the diabetic control in comparison to the normal rats(Graphs-6). Both doses of CSAE showed asignificant reduction in TC level. CSEE of 200mg/kg showed a significant reduction in the TG levels. CSAE (400mg/kg), CSEE(200mg/kg) and CSEE(400mg/kg) showed a significant elevation of HDL levels in comparison to the normal and diabetic control. Both extracts of all the doses significantly reduced the LDL levels in comparison to diabetic control.
AI and CRI values are significantly higher in diabetic control compared to the normal rats (Graphs-2). Standard drug and test extracts significantly reduce the AI and CRI values. Glibenclamide, CSAE1, CSAE2, CSEE1 and CSEE2 showed 86.5, 64.6, 89.3,94 and 94.5 % protection respectively (Graphs-3).
The serum lipid levels are generally high in diabetes; mapping a major risk factor for coronary heart disease.6 Excess levels of TC and LDL are major coronary risk factors. The C. swietenia leaf extractreduced the TC, TG and LDL levels, where as it increased the cardioprotective lipid HDL levels significantly. It has been proved that raise in HDL levels is associated with a reduction in coronary risk.28In the present study, it has been observed that the C. swietenia leaf extract mitigated the raised TC and LDL levels in diabetic rats. Further, it has been indicated that TG itself is independently linked to coronary heart disease29and in the present study, the plant extracts lowered TG levels in diabetic rats. The atherogenic index and the coronary risk index were very high in the diabetic rats.28Standard drug and plant extracts significantly reduced the AI, CRI as to the normalrats. % protection was increased with the dose, CSEE has shown more protection than the CSAE.
Diabetes is one of the common causes for a liver disease which includes abnormal liver enzymes, cirrhosis, hepatocellular carcinoma and acute liver failure.The AST, ALT, ALP , TB, and DB levels were raised in lever injury.27 These enzymes are considered as asensitive indicator of liver injury.Chloroxylonswietenia leaf extracts reduced the AST level, it shows the protective effect on theliver. The rise in ALP levels,indicates bone disease, bile tract obstruction or liver disease. C. swieteniaextracts lowered the ALP levels, suggesting its protective effect on liver function.
Diabetes affects the kidney, result in the development of diabetic nephropathy. Serum creatinine levels reveal the kidney function.30CSEE (200mg/kg) significantly reduced the creatinine levels.
It is evident that C. swietenia leafextracts contain antihyperglycemic agents capable of reducing the blood glucose level.
The authors are thankful to the colleagues and management for facilitating to accomplish this project.
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