HYPOGLYCEMIC EFFECT OF ACALYPHA INDICA LINN. PLANT EXTRACTS ON STREPTOZOTOCIN INDUCED DIABETES IN RAT

HYPOGLYCEMIC EFFECT OF ACALYPHA INDICA LINN. PLANT EXTRACTS ON STREPTOZOTOCIN INDUCED DIABETES IN RAT

Rajsekhar Saha* and Azhar Ahmed

Department of Pharmacology, RKDF College of Pharmacy, Hoshangabad Road, Bhopal, 462026, Madhya Pradesh, India

ABSTRACT

The plant Acalypha indica Linn.  is commonly known  as Indian Acalypha  and it belongs to the family  Euphorbiaceae, found in all parts of the tropics. This herbs found in fields and wastes places throughout the hotter parts of the world. The plant has wide uses in the traditional   medicines of   various countries and reportedly possesses diuretic, purgative and anthelmintic properties, besides being also used for bronchitis, asthma, pneumonia, scabies and other cutaneous diseases. In the present study different extract  of  Acalypha indica plant was evaluated  on  fasting  and  posts  prandial  blood sugar in both insulin and non insulin dependent diabetes  mellitus  rats. The induction of diabetes was done by using streptozotocin in neonates. Oral administration of petroleum ether, chloroform, acetone and methanol extracts of the bark (100 mg/kg, p.o.) for 21 days caused a decrease in fasting blood sugar (FBS) in diabetic rats. Among all the extracts, methanol extract was found to lower the FBS significantly (P<0.001) in diabetic rats. Methanol extract also caused a significant decrease (P<0.01) in post prandial blood sugar level in diabetic rats. Results of the test were compared with the standard antidiabetic drug glibenclamide (5mg/kg, p.o). Thus the   present study indicates that the methanol extract of Acalypha indica possess antidiabetic activity.

Streptozotocin

INTRODUCTION: Diabetes mellitus is a heterogeneous group of disorders characterized by abnormalities in Carbohydrate, protein, and lipid metabolism⁴. The central disturbance in diabetes mellitus is an abnormality  in  insulin  production  or  action  or  both,  although  other  factors can be involved.  Hyperglycemia is a common end point for all types of diabetes mellitus and is the parameter that is measured to evaluate and manage the efficacy of diabetes therapy.

Diabetes mellitus occurs in several forms, approximately 10% of diabetes patients have type 1 diabetes mellitus, an autoimmune disease that destroys insulin- producing beta cells in the pancreas leading to decrease in the concentration of insulin in the body and the  remainder have type 2 (Non insulin dependent diabetes mellitus).

Type 2 diabetes mellitus is a metabolic disorder characterized by a progressive decline in insulin action (insulin resistance, followed   by   the   inability of   pancreatic ß cells to compensate for insulin resistance) normally compensate for insulin resistance by secreting greater amount of insulin needed to maintain glucose homeostasis ⁵. In non insulin dependent diabetes mellitus this cell function  becomes  impaired  due  to  insulin  resistance leading  to deterioration in glucose homeostasis and  subsequent development of impaired glucose tolerance.

Diabetes mellitus is recognized as a major risk factor for cardiovascular diseases (CVD) such as atherosclerosis, heart attack, stroke, etc. About 75% of deaths among men with diabetes and 57% among women with diabetes are attributable to CVD. Many medicinal plants have been evidenced for having anti diabetic activity having lipid lowering character. Ethnobotanical information also indicates that more than 800 plants are used as traditional remedies for the treatment of diabetes throughout the world.

Acalypha indica Linn. is an annual erect herb  30-75 cm in height. Branches are numerous, long, ascending,   finely pubescent.  It is commonly known   as Indian Acalypha and it belongs to the family Euphorbiaceae, found in all parts of the tropics. This herbs found in fields and   wastes places throughout   the   hotter parts of the world. The plant has wide uses in the traditional medicines of various countries and reportedly possesses diuretic, purgative and    antihelmintic properties, besides being also used for bronchitis, asthma, pneumonia, scabies and other cutaneous   diseases ^{3, 6}.

The activities delivered by the plant reported are analgesic, Anti-inflammatory, Antihelmintic activity, Anti bacterial and anti fungal activity, Anti tubercular activity, antioxidants ^8-19. The  plant  reported  to  consist of Alkaloids, Catachols,  Flavonoids,  Phenolic  compounds, Saponins,  Steroids,  Tannins ⁷. Therefore, the present study was conducted to evaluate the sugar lowering potential of bark extract of this plant on streptozotocin induced NIDDM rats.

MATERIAL AND METHODS:

Plant and Chemicals: Acalypha indica plants were collected from the forest of Raisen district of Madhya Pradesh in   the   month of    august. The plant was identified and authenticated and deposited in the department. Streptozotocin   was   purchased   from   Calbiochem,    Germany.     Standard antidiabetic drug   glibenclamide was obtained from Ranbaxy Research Laboratories, Gurgaon, India. Analytical grade chemicals including various organic solvents (petroleum ether, chloroform, acetone and methanol) from E. Merck India Ltd., and Ranbaxy Laboratories India, were used for the extraction and phytochemical study of the constituents.

Preparation of Different Plant Extracts: Whole  plant  was  shade  dried  at  room temperature, ground into fine powder and then extracted  (amount 450g)  with    solvents   of   increasing     polarity    such as petroleum ether, chloroform, acetone   and  methanol  for  24h with each solvent by hot extraction using Soxhlet apparatus  at  a  temperature of 60ºC. The   extracts   were concentrated under reduced pressure using a rotary evaporator to constant weight and collected and preserved in desiccator for further studies.

Phytochemical Study: A portion of residue from each extract was subjected to phytochemical analysis in order to see the presence of sterols, alkaloids, carbohydrates, tannins, phenols, etc. in the plant extracts ²⁰.

Determination of Blood Sugar Level in Diabetic Rats:

Animals: Wistar albino rats of either sex were randomly bred in the Institutional animal house. The   animals   were   housed   in   standard   polypropylene  cages and maintained under controlled room   temperature (22^o+2^oC)  and  humidity (55+5%) with 12:12 h light and dark cycle. All the animals were provided with commercially available rat normal pellet diet and water ad libitum. The guidelines of the Committee for the Purpose of Control and Supervision of     Experiments on Animals (CPCSEA) of the Govt. of   India were followed and prior permission and   clearance were obtained from the institutional animal ethics committee for conducting the animal experiments.

Induction of diabetes: Diabetes mellitus was induced in five day old neonates (50 animals) by intraperitoneal injection of streptozotocin (90 mg/kg in 0.1M citrate buffer pH 4.5). The control group received equivalent amount of citrate buffer. The animals were allowed to live with their respective mothers and weaned from breast feeding at 4 weeks of age. Eight weeks after injection of streptozotocin, the rats were checked for fasting blood sugar (FBS) level by glucose oxidase-peroxidase method. Animals showing FBS more than 150 mg/dl were considered as diabetic (38 animals) and included for the study ²¹.

Treatment protocol: The diabetic animals were divided into six groups, each containing six animals and one group of normal non diabetic animals. All the extracts of A. indica plant was given at a dose of 100 mg/kg, orally for a period of 21 days as a suspension in Tween 80 to different groups of diabetic animals.

Gr I: Normal animals received Tween 80 in a dose of 1% suspension in distilled water.

Gr II: Diabetic animals received Tween 80 in a dose of 1% suspension in distilled water.

Gr III: Diabetic animals received standard antidiabetic drug glibenclamide (5mg/kg, p.o.)

Gr IV: Diabetic animals received Petroleum ether extract (100mg/kg, p.o.)

Gr V: Diabetic animals received chloroform extract (100mg/kg, p.o)

Gr VI: Diabetic animals received acetone extract (100mg/kg, p.o)

Gr VII: Diabetic animals received methanol extract (100mg/kg, p.o.)

At  the end of the experimental period the animals were fasted overnight for 8h and blood was taken  from  the  retro orbital plexus under mild ether anesthesia, serum was separated out and blood  sugar  level  was  evaluated  by  the method of glucose oxidase-peroxides method using span diagnostic kits.

Antihyperglycaemic Assay:

Oral Glucose Tolerance Test: The    six    groups of   diabetic animals and one group of normal animals were fasted overnight. Hyperglycaemia  was  induced  by  oral administration of 75% glucose solution .The standard  oral  glucose  tolerance  test  was  performed  on  all animals before and after giving the treatment²¹.

Statistical analysis: The results were expressed as Mean ± SEM. The unpaired t-test was used for analyzing the   data between two groups. Statistical   analysis of data was initially performed by using analysis of variance (ANOVA) when  the  overall ANOVA was significant, unpaired t-test was applied to study the difference among the groups.

RESULTS AND DISCUSSION:

Phytochemical study: The whole   plant   extract   of   Acalypha indiac Linn. was assayed for the presence of Anthroquinone, Alkaloids, Catachols, Flavonoids,   Phenolic compounds, Saponins, Steroids, Tannins, Triterpenoids.

The    result   concluded   the   presence of Alkaloids, Catachols, Flavonoids, Phenolic compounds, Saponins, Steroids ⁷.

Effect on fasting blood sugar and post prandial Blood Sugar Level of Diabetic Rats:    The  effect  of  different  extracts  on  serum  glucose  level  in  the diabetic rats shown in Table 1 revealed that all the extracts caused  reduction in blood glucose level but maximum reduction was found.

TABLE 1: EFFECT OF A. INDICA PLANT EXTRACTS ON FASTING BLOOD SUGAR OF DIABETIC RATS (n=6)

Results are expressed as Mean ± SEM. Results of the test and standard groups are compared with the control group. *P<0.05, **P<0.01, ***P<0.001

In the group treated with methanol extract. Methanol extract showed 51% reduction (P<0.01) as compared to glibenclamide which showed 67% reduction in fasting blood sugar. Table 2 summarizes the effect of the plant extracts on post prandial sugar level. There is no significant decrease in the glucose level in animals   without treatment. A. indica and glibenclamide   treatment   caused   significant   decrease   in   blood sugar level after 2 hours. Administration of streptozotocin caused rapid destruction of pancreatic ß cells in rats, which led to impaired glucose-stimulated insulin release and insulin resistance, both of which are marked feature of type II diabetes. The hypoglycaemic effect of plant extracts is generally dependent upon the degree of pancreatic ß cell   destruction and useful in moderate streptozotocin   induced   diabetes. The lesser the degree of pancreatic ß cells destruction, the more useful the herb is in treating diabetes in animals.

TABLE 2: EFFECT OF A. INDICA PLANT EXTRACT ON POST PRANDIAL BLOOD SUGAR LEVEL OF DIABETIC RATS (n=6)

Results are expressed as Mean ± SEM. Results of the test and standard groups are compared with the control group. *P<0.05, ** P<0.01, ***P<0.001.

CONCLUSION: Among all the extracts tested, methanol extract caused significant reduction in the serum   blood   glucose   level   as   compared to glibenclamide. Methanol extract caused 51% reduction in FBS level in diabetic animals whereas glibenclamide caused 67% reduction in sugar level. Methanol   extract also caused   a significant (P<0.01) decrease in post prandial blood sugar level in diabetic animals.

Preliminary phytochemical screening revealed the presence of    Alkaloids, Catachols, Flavonoids, Phenolic compounds, Saponins,   Steroids. Estimation of insulin level and insulin receptor may give more insight into the mechanism of its antidiabetic action. There is ongoing research to isolate and characterize the bioactive compound(s) responsible for the antidiabetic activity of A. indica.

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