Preparation and Evaluation of Novel Antidiabetic Polyherbal Formulation

 

Santosh Kumar Mahapatra1, Seema Verma2*

1Institute of Pharmacy and Technology, Salipur, Cuttack.

2Royal College of Pharmacy, Raipur, Chhattisgarh.

*Corresponding Author E-mail: sv729938@gmail.com

 

ABSTRACT:

Polyherbal formulation having more than one plant in the particular dosage form. These formulation shows greater therapeutic efficacy compared to single plant. In the present study. It was intended to formulate antidiabetic polyherbal formulation incorporating different ratio of the Annona squamosa (leaves), Withania somnifera (roots), Tinospora cordifolia (stems)and Azadirachta indica (leaves). The seven different polyherbal formulation (HF1 to HF7) were prepared and administered orally to the streptozotocin induced diabetic rats. The experimental rats treated with the HF1 to HF7 and reference drug Glibenclamide significantly reduced the blood glucose level compared to the diabetic control group. Further, the HF1 to HF7 showed antihyperlipidemic activity by lowering the total cholesterol, triglycerides and low-density lipoprotein level as well asenhanced the high-density lipoprotein. The polyherbal formulations and reference drug improve the body wights of the rats. The HF6 demonstrated greater antidiabetic and antihyperlipidemic activity compared to other polyherbal formulations.

 

KEYWORDS: Polyherbal formulation, antidiabetic, antihyperlipidemic, streptozotocin.

 

 


INTRODUCTION:

The astronomic increment in the commonness of diabetes has made diabetes a noteworthy general wellbeing challenge for India and is turned out to be significant human disease harassing numerous from different backgrounds in various nations and indeed the entire world being viewed ayurvedic the most established recuperating arrangement of medication for the treatment of diabetes. In spite of the fact that there are numerous synthetic medicines developed for patients, yet the reality it has never been accounted for that somebody had recouped absolutely from diabetes. The latest oral hypoglycemic medications are associated with a number of unwanted and adverse consequences. So much effort has been put into considering the antidiabetic properties of medicinal plants and herbal formulations in the treatment of disease in recent years that it is now a major focus of research1,2.

 

When it comes to Ayurvedic medication details, two things must be considered: Usage as a single medication or in combination with other treatments, the latter being known as Polyherbal formulation. In this fundamental traditional curative herbal technique, which is also known as polypharmacy or polyherbalism, the combining of a handful medicinal herbs in order to obtain more beneficial effectiveness is exploited to create greater beneficial viability3,4.

 

According to the Ayurvedic text Sarangdhar Samhita, polyherbalism can be used to create increasingly evident therapeutic sufficiency over time. Individual plants do not contain enough active compounds to produce the desirable and beneficial effects that are desired. Whenever the numerous plants are combined to a specific extent, this will have a greater beneficial effect and will reduce the toxicity of the combination5.

 

Science has found the fact that when herbs of various strength are mixed, the result may theoretically be more significant than whenever the plants are used individually, and that the sum of their separate effects could theoretically be more significant than when the plants are used individually. Synergism is the term used to describe the phenomena of beneficial herb-herb interplay. A number of the pharmacological actions of botanical active compounds are notable primarily when they are potentiated by the effects of different herbs, but are not noticeable if they're used alone. For example, combining ginger with dark pepper and long pepper enhances therapeutic efficacy. Black pepper, Cuminand asafoetida are historically combined to reduce edoema caused by poor digestion, while guduchi and turmeric enhance immunity6,7.

 

Polyherbalism offers several benefits that aren't available in individual herbal formulations due to synergistic effects. It is self-evident that a single multi formulation will have a greater therapeutic effect. In order to achieve enticing pharmacological activity, a lesser dose of the herbal concoction would be required, hence minimising the likelihood of adverse side-effects. Furthermore, polyherbal formulations enhance patient comfort by eliminating the requirement to take more than one distinct single herbal formulation at the same time, that results in greater consistency and curative impact as a result of this. When compared to single herbal formulation, each of those benefits has contributed to the increased popularity of polyherbal formulation on the trade8,9.

 

As a result, it was decided to create an anti-diabetic polyherbal polyherbal formulation containing different ratio of containing Annona squamosa(leaves),Withania somnifera(roots), Tinospora cordifolia(stems)and Azadirachta indica(leaves).

 

MATERIAL AND METHODS:

The leaves of Annona squamosa and Azadirachta indica, roots of Withania somnifera and stems of Tinospora cordifolia were collected, after cleaning plants parts were shade dried. The plant parts were further processed for the coarsely powdered and kept in air tight container for experimental work.

 

Preparation of Polyherbal Formulations:

There were a total of seven distinct polyherbal formulations (HF1, HF2, HF3, HF4, HF5, HF6 and HF7) created by blending varying ratios of plant powders of Annona squamosa (leaves), Withania somnifera (roots), Tinospora cordifolia (stems)and Azadirachta indica (leaves). Table 1 lists the constituents of various polyherbal formulations in varying proportions.

 

Table 1: Polyherbal formulations are composed of a variety of herbs

Formulations

Ratio

Annona squamosa

Withania somnifera

Tinospora cordifolia

Azadirachta indica

HF1

1

1

1

1

HF2

1

2

1

1

HF3

1

2

2

1

HF4

1

2

2

2

HF5

2

1

2

2

HF6

2

1

1

2

HF7

2

1

1

1

 

Decoction Preparation:

The mixtures of 20 g of every composition with 150 ml of distilled water have been macerated for 24 hours at room temperature. Decoction were obtained by boiling about 45 minutes then filtering with muslin cloth the drug macerate that had been left for 24 hours. Adjustments were made to the decoction's content so that 20 g of mixture yielded 50 ml of decoction10.

 

Antidiabetic Activity:

Polyherbal Formulation's Oral Glucose Tolerance Test:

The oral glucose tolerance test was carried out in the experimental rats that had been fasted for18 hours. The rats were placed into ten groups for the experiment, and each group have six rats. Group I named as a normal control, group II have glucose control rodents, group III treated with reference drug Glibenclamide(0.5 mg/kg), while group IV to Group X treated with HF1 to HF10, respectively at the dose of 20 ml/kg body weight. 

 

Thirty minutes before the administration of the extracts and the reference medication, rats in Groups II to X were given glucose (2 g/kg). Blood being taken from the retro-orbital sinus at 0, 30, and 90 minutes following introduction of the extract and reference medication. The plasma obtained after centrifugation of blood at 3000 rpm, was used to assess fasting plasma glucose levels using a glucose oxidase–peroxidase kit11,12.

 

Non-Insulin-Dependent Diabetes Mellitus Onset:

A virtually overnight starved adult rat weighing 170–220 g were used to develop non-insulin dependent diabetes mellitus (NIDDM). A single intraperitoneal injection of 60 mg/kg Streptozotocin 15 minutes after administering 120 mg/kg nicotinamide to develop NIDDM in these rats. The presence of diabetes was verified by the appearance of raised glucose levels in plasma, that were measured at 72 hours and again on day 7 after administration. The fasting plasma glucose level of > 126 mg/dl was established as the diagnostic threshold for diabetes. Specifically, those rodents being employed in the experiment who were confirmed to be have persistent NIDDM.

 

Antidiabetic Activity of Polyherbal Formulation:

The rodents have been divided into 10 groups of six rats each, for a total of 120 rats. The polyherbal formulation and reference drug was given to the animals for a total of 28 days. Group I named as a normal control administered only drinking water, group II have diabetic control rodents, group III treated with reference drug Glibenclamide (0.5 mg/kg), while group IV to Group X treated with HF1 to HF10, respectively at the dose of 20 ml/kg body weight for 28 days.

 

The blood glucose levels were measured on the first, seventh, fourteenth, and twenty-eighth days after the extract administration. Over the course of the study, the rodents have been weighed on a regular basis, and the average difference in body mass was computed13,14.

 

Estimation of Biochemical Parameters:

It was on day 28 that the biochemical variables was measured afterwards the rats were sacrificed through spinal displacement. The glucose oxidase technique was used to quantify total cholesterol, triglycerides (TGL), high-density lipoprotein (HDL), and low-density lipoprotein (LDL) employing an auto-analyzer15,16.

 

Statistical Analysis:

The data are presented as the average of six independent experiments with standard deviations (SEM). The statistically significant difference of the variations were observed was determined using one-way analysis of variance (ANOVA) and Dunet's test. A p-value of less than 0.05 was deemed statistically significance in this study.

 

RESULTS AND DISCUSSIONS:

Effects of polyherbal formulation on oral glucose tolerance

Table 2 illustrates the impact of polyherbal formulations on the amount of glucose in the blood plasma of animals. Following the ingestion of sugar, an increase in blood glucose was detected in the glucose control, polyherbal formulation (HF1 to HF7) medicated, and referece group animals. When comparing the animals treated with polyherbal formulation to the control group, a substantial drop in hyperglycaemia was noted. The glibenclamide-treated group also experienced a substantial reduction in plasma glucose levels, according to the findings. The polyherbal formulation showed glucose tolerance efficacy in following orders HF6 > HF3 > HF2 > HF4 > HF7 > HF5 > HF1. The HF6 has greatest glucose tolerance properties compared to other polyherbal formulation.

 

Table 2: Effect of oral glucose tolerance test of polyherbal formulationson rats

Group

Blood glucose (mg/dl)

0 min

30 min

90 min

Normal Control

73.1±5.3

76.7±6.5

75.5±3.8

Glucose control

79.4±3.2

214.3±2.1a

153.4±2.2a

Glucose + Glibenclamide

77.3±5.6

109.7±2.5*

77.2±5.2*

HF1

78.2±4.1

159.1±5.3*

90.6±1.4*

HF2

74.8±6.4

118.1±4.1*

77.2±5.4*

HF3

77.6±1.8

112.3±7.1*

75.9±7.2*

HF4

73.1±5.1

122.4±3.6*

79.5±3.3*

HF5

76.4±3.8

136.2±7.1*

85.1±6.5*

HF6

75.2±2.3

108.6±6.1*

75.1±4.2*

HF7

78.5±4.2

129.7±3.5*

80.6±3.7*

Findings are demonstrated as mean ± SEM (n=6); statistically difference at aP<0.05once comparing with the normal control group, and *P<0.05 once comparing with the diabetes control group

 

Effect on Niddm of Polyherbal Formulation:

The observation of elevated plasma glucose levels in glucose treated rats validated the establishment of hyperglycemia in these animals. On the other hand, the efficacy of a HF1 to HF7 on serum glucose levels in normal and STZ-induced diabetic rodents is demonstrated in Table 3.

 

When comparing the STZ treated diabetic control rodents to the normal control rodents, a substantial increased serum glucose levels was seen on the first, seventh, fourteenth, and twenty-eighth days after treatment with streptozotocin. A considerable reduction in serum glucose levels was observed in the rodents administered with theHF1 to HF7 andreference drug glibenclamide (0.5 mg/kg p.o.) as compared to STZ induced diabetic rats. When HF6 is compared to other polyherbal formulations, the findings show that HF6 has more significant antidiabetic effects.

 

Table 3: Antidiabetic effect of polyherbal formulationsin STZ treated rodents

Groups

Blood glucose (mg/dl)

0Day

7thDay

14thDay

28thDay

Normal Control

80.3±7.2

77.1±4.2

79.6±4.7

80.7±4.5

Diabetic control (Streptozotocin)

153.8±2.5a

198.2±6.7a

254.7±6.3a

291.3±5.3a

Glibenclamide

152.1±4.2

104.7±5.7*

87.2±3.8*

72.8±5.1*

HF1

163.4±5.7

149.8±5.3*

121.7±4.5*

96.9±4.3*

HF2

154.7±5.3

119.1±5.1*

92.5±4.3*

78.7±5.9*

HF3

149.7±1.8

108.6±6.2*

90.3±5.2*

78.2±4.3*

HF4

160.2±3.5

121.7±2.8*

98.4±6.8*

79.6±7.5*

HF5

139.7±6.7

141.4±4.1*

116.9±5.1*

90.4±7.2*

HF6

143.5±5.8

106.5±4.2*

88.1±6.7*

73.9±7.5*

HF7

151.8±4.2

132.1±4.2*

109.7±3.2*

83.1±4.5*

Findings are demonstratedas mean ± SEM (n=6); statistically difference at aP<0.05once comparing with the normal control group, and *P<0.05 once comparing with the diabetes control group

 

Anti-Hyperlipidaemic Activity of Polyherbal Formulation:

As shown in Table 4, the results of the blood lipids of control and experimental rodents were comparable. As comparison to normal rodents, diabetic control animals had a large rise in total serum cholesterol, LDL cholesterol, and HDL cholesterol, despite having a substantial decrease in HDL cholesterol. When comparing to the diabetic control group, the rodents administered with glibenclamide and polyherbal formulations had lower TGL, lower total cholesterol, lower LDL, and higher HDL. All of such impacts were found on the 28th day of the experiment. When HF6 was contrasted to other polyherbal formulations, the results of the blood lipids revealed that HF6 has the greatest antihyperlipidemic effect.

 

Table 4: Biochemical parameters following administration of polyherbal formulations

Group

Biochemical Lipid Profile

TGL (mg/dl)

TCL (mg/dl)

LDL (mg/dl)

HDL (mg/dl)

Normal control

71.3±3.9

79.5±7.1

61.4±1.9

82.7±6.6

Diabetic control (Streptozotocin)

225.1±6.1a

196.6±5.9a

176.8±7.5 a

22.9±7.5 a

Glibenclamide

72.8±5.6*

73.9±3.2*

65.4±4.7*

84.1±2.6*

HF1

93.7±4.9*

90.1±4.6*

88.3±2.6*

60.3±5.8

HF2

74.6±5.3*

79.4±2.5*

65.3±4.9*

80.6±6.5*

HF3

73.7±5.3*

75.8±7.3*

64.2±5.8*

83.6±2.1*

HF4

74.2±4.6*

80.1±7.2*

68.1±5.5*

77.9±4.9*

HF5

89.1±6.5*

86.2±3.5*

81.7±1.8*

68.5±2.9*

HF6

73.2±6.3*

74.1±1.7*

63.9±3.8*

85.7±7.5*

HF7

77.6±6.2*

83.9±2.4*

75.8±7.9*

72.3±6.5*

Findings are demonstratedas mean ± SEM (n=6); statistically difference at aP<0.05once comparing with the normal control group, and *P<0.05 once comparing with the diabetes control group; Triglyceride-TGL; Total Cholesterol- TCL; low-density lipoprotein-LDL;high-density lipoprotein; high-density lipoprotein-HDL

 

Effect of Polyherbal Formulation on Rats Body Weight:

Throughout the course of the investigation, the body weights of rodents have been determined before and after production of hyperglycemia, including afterwards treatment (Table 5). Rats' body weight reduced after being diagnosed with diabetes, and their weight recovered after being treated with polyherbal formulations, according to the findings of the study.

 

Table 5: Variations in body weight of rodents using polyherbal formulations

Group

Body weight of rats

Before Initiation (gm)

After Induction of diabetics(gm)

After Treatment(gm)

Normal control

189.2±5.3

185.7±4.5

186.3±6.2

Diabetic control (Streptozotocin)

182.4±4.8

121.6±7.5

115.1±4.6

Glibenclamide

179.5±2.8

140.2±4.1

180.9±3.7

HF1

185.3±5.2

146.7±3.5

172.5±4.8

HF2

172.1±6.3

141.2±5.5

173.4±3.2

HF3

180.9±2.5

137.4±4.2

178.2±5.2

HF4

176.7±4.9

139.3±5.4

177.5±6.7

HF5

183.8±5.2

143.1±5.2

180.4±6.7

HF6

178.1±3.5

146.7±5.2

176.2±5.7

HF7

188.6±6.2

152.5±4.5

185.3±6.7

STZ is indeed a poisonous glycoside derived from the bacterium Streptomyces achromogenes. It is used to treat a variety of diseases. It aggregates in pancreas cells through the glucose transporter 2 (GLUT2) and suppresses the production of the enzymes involved. The alkylating characteristics of the STZ cause cellular components to be modified, DNA to be fragmented, and cell to be destroyed, resulting in insulin-dependent hyperglycemia. In the diabetes control group, substantial weight loss has been noted, that may have been caused by enhanced muscles wastage and protein loss from the body's cells. The current study revealed revealed the treated group experienced considerable weight loss, indicating that the polyherbal formulation and glibenclamide are effective in preventing hyperglycemia-induced muscle loss. In other cases, a decrease in glucose concentrations might be attributed to an increment in plasma insulin levels or an enhancement in the transit of glucose into peripheral tissues17,18. Our findings demonstrate that the polyherbal formulation lowers plasma glucose levels and also shows promise antidiabetic efficacy, which is consistent with previous research.

 

The chronic hyperglycemia caused by STZ results in changed plasma biochemistry levels of Total Cholesterol, Triglyceride, LDL, and HDL, each of that are regarded to be major indicators of diabetes mellitus and mortality. The polyherbal formulation that was administered to the rats had the ability of reversing the effects of STZ on the lipid profile. It could be attributable to the specific plants in the polyherbal formulation's anti-diabetic function.

 

CONCLUSION:

In the present study, the polyherbal formulation namely HF1 to HF7 incorporating different ratio of the Annona squamosa (leaves), Withania somnifera (roots), Tinospora cordifolia (stems)and Azadirachta indica (leaves) were prepared for the evaluation of the antidiabetic activity. The HF1 to HF7 significantly reduced the blood glucose level in the diabetic rats and also monitor the lipid profile. The findings suggested, HF6 having higher antidiabetic and antihyperlipidemic activity compared to other formulation. Looking on antidiabetic efficacy of the polyherbal formulation, planned to carry out the safety profile study in the future. Also this study has scope to illustrate the possible mechanism of antidiabetic activity of this polyherbal formulation.

 

CONFLICT OF INTEREST:

None

 

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Received on 09.05.2022           Modified on 08.06.2022

Accepted on 29.06.2022         © RJPT All right reserved

Research J. Pharm. and Tech. 2022; 15(7):3015-3019.

DOI: 10.52711/0974-360X.2022.00503