Antidiabetic activity of Polyherbal formulations from Chhattisgarh State

 

Amit Roy, Pushpa P. Gupta, Shiv Bharadwaj, Shashikant Chandrakar

Columbia Institute of Pharmacy, Raipur (C.G.)

*Corresponding Author E-mail: pushpaprasad81@gmail.com

 

ABSTRACT:

Diabetes mellitus is described as a metabolic disorder of multiple etiologies characterized by insulin resistance, relative insulin deficiency and hyperglycemia with disturbances of carbohydrate, fat and protein metabolism. It is predicted that by 2030 diabetes mellitus may affect up to 79.4 million individuals in India. In the present study four polyherbal formulations were collected from various places of Chhattisgarh state, India. These formulations were selected on the recommendation of local healers. The formulations were designated as F1, F2, F3 and F4. These formulations were administered to the rats in two different doses i.e. 200 and 400mg/kg body weight; glibenclamide was used as standard drug. The antidiabetic effect of these polyherbal formulations was studied in streptozotocin induced diabetic rats. The blood glucose level of diabetic rat significantly increased compared to normal rats. Maximum reduction was seen in animals treated with F4 at its both the doses. All the formulations significantly lowered the levels of cholesterol, triglycerides and LDL and at the same time HDL level increased in rats treated with extract and standard drug compared to untreated rats. The body weight of rats belonging to diabetic control group decreased significantly on induction of diabetes. The rats treated with extract and glibenclamide were found to gain body weight when compared to diabetic control group. It is interesting that in polyherbal formulation treated rats there was significant decrease in blood glucose level and also lipid peroxidation;  the observation shows increased levels of the antioxidant enzymes and decrease in lysosomal enzymes. The potent antidiabetic effect of the polyherbal formulation suggests the presence of such phytoconstitutents that can produce significant antihyperglycemic activity as well as other biochemical changes that can protect from complications of diabetes. The result of present study also suggests that the claims made by the local healers about the efficacy of formulation is correct, however extensive study is required to determine the mechanism by which the activity take place and at the same time the biological markers present in the formulations that are responsible for the activity.

 

KEYWORDS: Antidiabetic activity, polyherbal formulation, Chhattisgarh.

 

 

INTRODUCTION:

Diabetes mellitus describes a metabolic disorder of multiple etiologies characterized by insulin resistance, relative insulin deficiency and hyperglycemia with disturbances of carbohydrate, fat and protein metabolism. India is one among the leading country with highest number of diabetic population with a current figure of 40.9million, followed by China, USA, Russia, Germany, Japan, Pakistan, Brazil, Mexico and Egypt. The etiology of diabetes in India is multifactorial and includes genetic factors coupled with environmental influences such as obesity associated with rising living standards, steady urban migration, and lifestyle changes1,2. It is predicted that by 2030 diabetes mellitus may afflict up to 79.4million individuals in India. Diabetes has become a serious health problem with continuously increasing rates of incidence and mortality. The management of diabetes mellitus is considered a global problem and successful treatment is yet to be discovered. Even though the synthetic drugs, including insulin and oral hypoglycemic agents, control the blood sugar level as long as they are regularly administered, Cost, Complications, limited tolerability and other side effects reduces its wide acceptance. This situation may be the main reason for the shift of common people form allopathic system to herbal formulation nowadays3.

 

In traditional systems of medicine, many plants have been documented to be useful for the treatment of various systemic disorders. Many of the traditional/indigenous systems of medicine are effective than the modern system of medicine, but they suffer from lack of complete standardization which is one of the important challenges faced by the traditional system of medicine. The concept of polyhedral formulation is well documented in the ancient literature. Compared to the single herb, the polyherbal formulation has better and extended therapeutic potential.

 

The four different polyherbal preparations which is widely used in the effective management of diabetes by traditional healers of different places namely Baloudabazar, Mungeli, Amleshwar, Janjgir Champa of Chhattisgarh state. These formulations claimed to maintain associated symptoms such as polyuria, fatigue, constipation, dryness of mouth, polydipsia and excessive sweating. Further, scientific data regarding the antidiabetic effect of four formulations are lacking. Hence, the present study was planned to provide experimental basis to clinical findings, antidiabetic activity at primary level has been undertaken in streptozotocin induced diabetic rats.

 

MATERIAL AND METHODS:

Collection of polyherbal formulation:

The four different polyherbal formulations were obtained from various places of Baloudabazar, Mungeli, Amleshwar and Janjgir Champa of Chhattisgarh state, India. The details of polyherbal formulations are given in table 1. 

 

Table 1: List of antidiabetic polyherbal formulations collected for experimental purpose

Place of collection

Formulation code

Vill. Mudpar, tahsil- Sigma, Dist. Baloudabazar

F1

Vill. Dabari, Dist. Mungeli

F2

Amleshwar

F3

Janjgir Champa

F4

 

Preparation of extract

The Hydroalcoholic (70:30) extract of polyherbal formulations (F1 to F4) were prepared for further studies.

 

Antidiabetic activity: 

The streptozotocin induced diabetic rats model was performed to assess the antidiabetic activity of polyherbal formulation (F1 to F4).

 

Selection of animals:

Animal care and handling:

Studies were carried out using Male Wistar rats weighing 150 to 230gm. The rats were grouped and housed in polyacrylic cages (38 x 23 x 10cm) with not more than six animals per cage and maintained under standard laboratory conditions (temperature 25 ± 2oC) with dark/ light cycle (14 /10 h). They were allowed free access to standard dry pellet diet and water ad libitum. The rats were acclimatized to laboratory conditions for 10 days before commencement of the experiment. All experiments were approved by the institutional ethical committee and were performed according to the animal ethics committee guidelines.

 

Induction of non-insulin dependent diabetes mellitus (NIDDM):

In overnight fasted rats (150 – 230g), Non-insulin dependent diabetes mellitus was induced. Streptozotocin was used to induce diabetes. A single dose of streptozotocin was given at a dose of 60 mg/kg intraperitonally, 15 minutes after nicotinamide administration (120mg/kg i.p.). Nicotinamide was dissolved in normal saline whereas STZ was in citrate buffer at a pH of 4.5 for administration. The raised glucose level in plasma was determined at 72 hours and then on 7th day, after induction. This data confirmed hyperglycemia. The threshold value of fasting plasma glucose level was taken as >126mg/dl for diagnosis of diabetes. The rats which have permanent NIIDM were used only for the study4-6.

 

Experimental design

The animals were divided into seven groups, each group containing six animals (n = 6), and extract of polyherbal formulation were administered for 28 days.

Group I rats were administered drinking water daily, served as normal control rats;

Group II rats were Diabetic control rats;

Group III rats were administered standard drug Glibenclamide (0.5 mg/kg);

Group IV rats were administered sample F1 (200 mg/kg);

Group V rats were administered sample F1 (400mg/kg);

Group VI rats were administered sample F2 (200 mg/kg);

Group VII rats were administered sample F2 (400 mg/kg);

Group VIII rats were administered sample F3 (200 mg/kg);

Group IX rats were administered sample F3 (400 mg/kg);

Group X rats were administered sample F4 (200mg/kg);

Group XI rats were administered sample F4 (400mg/kg)

On 0, 7th, 14th and 28th days of extract administration, the fasting glucose levels were determined. During the experimental tenure, weights of rats were taken daily and mean change was calculated.

 

Estimation of biochemical parameters

The animals were sacrificed by cervical dislocation on 28th day for determining various biochemical parameters. The method of glucose oxidase was used to determine total cholesterol, triglycerides (TGL), high-density lipoprotein (HDL) and low-density lipoprotein (LDL). The kit used was kits of Triglycerides and Cholesterol (Minias Globe Diagnostic kit)7-9. Semi auto-analyzer was used for the analysis of sample.

 

Statistical analysis

The results are expressed as mean±SEM of six independent experiments. Statistical significance between the groups was evaluated by one-way analysis of variance (ANOVA) followed by Dunet’s test. A P < 0.05 value was considered as statistically significant.

 

RESULT:

Antidiabetic Effect of polyherbal formulation on hyperglycemia

The effect of F1, F2, F3, F4 and glibenclamide on serum glucose levels in normal, diabetic, and extract treated rats is presented in Table 2. The blood glucose level of diabetic rat significantly increased compared to normal rats. The rats treated with F1, F2, F3 and F4 at two different doses (200mg/kg and 400mg/kg) to STZ-induced diabetic rats exhibited significant reduction of blood glucose level. The maximum reduction of blood glucose in rats was observed on 28th days. The blood glucose level of F1 at the dose 200mg/kg and 400mg/kg group was found to be 109.2±4.5mg/dl and 88.9±3.2 mg/dl, respectively at the end of study. The F2 treated rats at the dose of 200mg/kg and 400mg/kg exhibited 116.5±5.7mg/dl and 84.9±4.5mg/dl blood glucose level, respectively. The F3 treated rats at the dose of 200 mg/kg and 400mg/kg demonstrated 113.6±4.8mg/dl and 86.5±4.1mg/dl blood glucose level, respectively. The F4 treated rats at the dose of 200mg/kg and 400mg/kg revealed 103.8±5.7mg/dl and 86.4±3.2mg/dl blood glucose level, respectively.

 

 

 

Table 2: Effect of polyherbal formulation on fasting plasma glucose level in rats

Group

Fasting plasma glucose concentration (mg/dl)

Day 0

Day 7th

Day 14th

Day 28th

Normal Control

77.2±2.6

81.3±5.1

76.7±3.6

78.4±6.5

Diabetic control (Streptozotocin)

142.5±5.1a

197.6±3.4a

271.2±2.7a

353.6±4.1a

Diabetic + Standard Glibenclamide (0.50 mg/kg)

136.7±6.2

116.2±4.7*

98.5±3.4*

82.9±5.2*

Diabetic  + F1 (200 mg/kg)

145.2±3.1

141.5±2.8*

121.5±6.7*

109.2±4.5*

Diabetic  +  F1 (400 mg/kg)

131.8±5.6

126.5±3.9*

103.2±4.2*

88.9±3.2*

Diabetic  + F2 (200 mg/kg)

138.1±4.3

152.7±6.7*

127.3±3.9*

116.5±5.7*

Diabetic  +  F2 (400 mg/kg)

142.7±3.7

128.1±5.3*

97.2±2.6*

84.9±4.5*

Diabetic  + F3 (200 mg/kg)

147.2±2.4

146.8±2.9*

122.3±5.1*

113.6±4.8*

Diabetic  +  F3 (400 mg/kg)

139.5±3.1

122.5±4.7*

99.1±6.4*

86.5±4.1*

Diabetic  + F4 (200 mg/kg)

130.9±4.2

139.2±4.8*

118.6±5.6*

103.8±5.7*

Diabetic  +  F4 (400 mg/kg)

135.8±5.6

121.4±6.5*

96.1±3.8*

86.4±3.2*

Values are expressed as mean ± SEM (Number of animals, n=6); significantly different at aP<0.05 when compared with normal control group, *P<0.05 when compared with diabetic control group

 

Table 3: Determination of biochemical parameters after treatment with polyherbal formulation

Group

Lipid Profile (mg/dl)

Triglyceride

Total Cholesterol

HDL

LDL

Normal control

93.2±2.7

88.6±5.2

79.2±2.3

53.7±3.8

Diabetic control (Streptozotocin)

213.6±4.1a

172.1±3.7 a

21.2±4.6a

148.7±2.6a

Diabetic + Standard Glibenclamide (0.50 mg/kg)

88.1±3.8*

81.5±4.3*

86.2±2.1*

55.4±5.1*

Diabetic  + F1 (200 mg/kg)

102.3±5.2*

96.1±4.9*

51.8±3.7*

79.5±3.2*

Diabetic  +  F1 (400 mg/kg)

94.6±3.8*

86.5±2.1*

83.7±4.7*

59.1±3.5*

Diabetic  + F2 (200 mg/kg)

98.3±2.7*

93.4±3.8*

53.6±5.2*

82.7±4.1*

Diabetic  +  F2 (400 mg/kg)

83.7±4.6*

84.9±5.1*

81.2±2.4*

62.4±3.6*

Diabetic  + F3 (200 mg/kg)

99.1±3.7*

95.4±4.7*

52.9±5.9*

76.3±4.2*

Diabetic  +  F3 (400 mg/kg)

85.8±4.9*

81.5±2.1*

78.3±4.3*

59.4±5.6*

Diabetic  + F4 (200 mg/kg)

96.4±3.4*

97.5±5.9*

59.2±2.5*

81.3±2.8*

Diabetic  +  F4 (400 mg/kg)

88.2±2.1*

83.1±3.8*

73.7±5.4*

55.6±4.5*

Values are expressed as mean ± SEM (Number of animals, n=6); significantly different at aP<0.05 when compared with normal control group, *P<0.05 when compared with diabetic control group

 

 

Table 4: Effect of polyherbal formulation on changes in body weight in rats

Group

Change in Body weight (gm)

Before Induction

After Induction

After Treatment

Normal control

202.4±2.3

198.5±1.8

208.3±2.8

Diabetic control (Streptozotocin)

192.1±1.4

168.3±2.6

132.6±3.5

Diabetic + Standard Glibenclamide (0.50 mg/kg)

207±3.4

157.2±1.7

210.5±2.1

Diabetic  + F1 (200 mg/kg)

193.7±1.9

152.7±2.4

191.2±4.2

Diabetic  +  F1 (400 mg/kg)

188.4±2.4

171.6±1.8

196.3±2.5

Diabetic  + F2 (200 mg/kg)

197.3±1.7

168.2±2.3

201.6±1.9

Diabetic  +  F2 (400 mg/kg)

212.6±3.2

179.3±1.7

209.1±3.5

Diabetic  + F3 (200 mg/kg)

193.5±2.5

174.2±4.1

197.8±2.7

Diabetic  +  F3 (400 mg/kg)

199.4±2.7

181.6±3.9

205.8±1.6

Diabetic  + F4 (200 mg/kg)

184.7±3.9

174.9±1.6

185.3±3.2

Diabetic  +  F4 (400 mg/kg)

207.1±2.5

181.6±3.2

202.9±2.8

Values are expressed as mean ± SEM (Number of animals, n=6)

 

Effect of polyherbal formulation on lipid profile:

The F1, F2, F3 and F4 (200mg/kg and 400mg/kg) significantly lowered the levels of cholesterol, triglycerides and LDL in diabetic rats when compared with the diabetic control group (Table 3). Consequently, the HDL significantly increased in groups treated with extracts and standard in comparison to diabetic control group.

 

Effect of polyherbal formulation on body weight

The body weight of rats belonging to diabetic control group decreased significantly on induction of diabetes. The extract and glibenclamide treated rats were found to gain body weight when compared to diabetic control group as shown in Table 4.

 

DISCUSSION:

As per reported data, in India the indigenous system of medicines and ethno flora comprises 2532 medicinal plants. In India, approximately 45,000 plant genus are available and lots of them have been deliberate for their healing properties. About 2000 figures are available in the literature and commonly 500 species are used by indigenous systems. The Indigenous system of medicines is very popular, but its uses are not desirable due to lack of scientific evidence of herbal medicines. Polyherbal formulations are used extensively in Indian traditional medicine system for the management of type 2 diabetes to get synergistic effects. In the present study, polyherbal formulation collected from various places of Baloudabazar, Mungeli, Amleshwar and Janjgir Champa of Chhattisgarh state, India was selected for antidiabetic studies owing to its reported antidiabetic activity by local healers.  

 

The findings of present study indicate the significant effect of polyherbal formulation on Streptozotocin induced type 2 diabetic animals. Streptozotocin, a monofunctional nitrosourea derivative, derives diabetogenic activity due to its ability to induce oxidative stress and damage in β-cells. Streptozotocin can selectively attack pancreatic β-cells by producing free radicals of oxygen, nitrogen monoxide, and reducing intracellular NAD and NADP, which are crucial for the electron delivery and energy metabolism in β-cells10.

 

The significant oral glucose tolerance observed with administration of polyherbal formulation by decreasing blood glucose level in animals. The blood glucose lowering effect may be due to a decrease in glucose absorption, increased peripheral glucose utilization and glycolysis, decrease in glycogenolysis and gluconeogenesis. This indicates that polyherbal formulation has got appreciable glucose absorption decrement and postprandial glucose lowering by slow absorption of carbohydrate and inhibits glucose transport in gut11.

 

The diabetes was induced on rats after administration of Streptozotocin. The polyherbal formulations significantly reduced the blood glucose level in Streptozotocin -induced-diabetic rats as compared to the diabetic control group. The possible mechanism by which polyherbal formulation brings about its hypoglycemic action in diabetic rat may be by potentiating the insulin effect of plasma by increasing either the pancreatic secretion of insulin from the existing beta cells or by its release from the bound form.

Increased lipid peroxidation seen in diabetic conditions is attributed to increased oxidative stress in the cells as a result of the depletion of antioxidant systems. In support to this view levels of both enzymatic antioxidants decreased and lysosomal enzymes increased in diabetic rats. It is interesting that in polyherbal formulation treated rats there was decrease in lipid peroxidation, increased levels of the antioxidant enzymes and decrease in lysosomal enzymes. This shows that polyherbal formulation can reverse all these abnormalities either by pancreatic or hepatic mechanism12, 13. The antidiabetic effect of the plyherbal formulation suggests the presence of phytochemicals present in polyherbal formulations, which needs further investigation.

 

 

 

ACKNOWLEDGEMENT:

The authors are thankful to Chhattisgarh Council of Science and Technology, Raipur, Chhattisgarh for the grant provided for this project and Columbia Institute of Pharmacy, Raipur, Chhattisgarh, for providing necessary man power and infrastructure for completion of this project.

 

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Received on 25.07.2020           Modified on 14.08.2020

Accepted on 29.09.2020         © RJPT All right reserved

Research J. Pharm. and Tech. 2021; 14(3):1375-1379.

DOI: 10.5958/0974-360X.2021.00245.6