POLYHERBAL PREPARATION AND COMPARATIVE STUDIES ON DIET-INDUCED HYPERLIPIDEMIA

Objective: The objective of this research article is to develop and evaluate polyherbal preparation and comparative studies on diet-induced hyperlipidemia. Methods: After the extraction, pharmacognostical and phytochemical screening was done. The lipid-lowering activity of polyherbal formulation (T1, T2, T3, T4, and T5) may be attributed to the phytoconstituents present such as alkaloids, carbohydrates, steroids, proteins, tannins, carbohydrates, flavonoids, phenols, glycosides, and triterpenes. In acute oral toxicity study, there were no behavioral changes seen up to 4 h and no mortality was observed up to the end of 24 h even at the maximum tested dose level of 2000 mg/kg per oral. It was considered maximum safe dose. Male and female albino rats weighing 150–200 g were used for the study. Hydroalcoholic extract of all plants was prepared having a dose of 2000 mg/kg. The doses were selected according to the Organisation for Economic Cooperation and Development guideline no. 425. The procedure was divided into two phases: Phase I (observation made on day 1) and Phase II (observed the animals for the next 14 days of drug administration). Animals received a single dose of 2000 mg/kg. After the administration of Healthcare Administration, food was withheld for 3–4 h. In case animal dies, we have to again perform the test for the determination of LD50. The study was conducted by measuring various parameters, namely, daily feed intake (g), water intake (ml), body weight (g), lipid profile high-density lipoprotein (HDL), low-density lipoprotein (LDL), Cholesterol (CHL) level (mg/dl), and blood glucose level (mg/dl). Results: Results showed a significant decrease in blood glucose level and serum lipid profile such as total cholesterol, LDL, and increasing serum HDL level, so could be useful in the treatment of hypolipidemia. Conclusion: Polyherbal formulations (T1, T2, T3, T4, and T5) have hypoglycemic activity and significantly improve lipid profile levels in diet-induced experimental rats.


INTRODUCTION
Hyperlipidemia is known as a condition of an increased level of serum total cholesterol (TC), low-density lipoprotein (LDL), very LDL (VLDL), and reduced high-density lipoprotein (HDL) [1]. Cardiovascular diseases (CVDs) are a major risk factor caused by hyperlipidemia. Hyperlipidemia disease has afflicted humankind since antiquity. Hyperlipidemia, or hyperlipoproteinemia, is defined as abnormally elevated levels of one or more of triglycerides, cholesterol, cholesterol esters, and phospholipids and plasma lipoproteins including VLDL and LDL, and reduced HDL levels. These lipoproteins are deposit in the interstitial space of arteries arising from aorta, restricting the blood supply to the heart. This phenomenon is known as atherosclerosis. Due to the privilege deposition of lipoproteins, blood supply to the heart gets blocked, and thus, myocardial infarction occurs, which is usually well known as heart attack.
Among ischemic heart disease and the high mortality rate, there is a strong relation. In addition, high plasma cholesterol levels cause more than 4 million deaths in a year. Secondary hyperlipidemia often mimics familial forms of hyperlipidemia and can have similar effects. Due to secondary hyperlipidemia, risk of premature atherosclerosis increased or, when happen with severe hypertriglyceridemia, may cause pancreatitis and other consequences of the chylomicronemia syndrome [2,3].

Qureshi et al.
From times of yore, several studies have shown that dietary modifications such as high-fiber diets, low-fat diets, and diets rich in flavonoids and phenolic acids can reduce metabolic syndrome risk factors. Statins and synthetic antioxidants such as probucol are modern antihyperlipidemic drugs which are widely used to treat atherosclerosis. Regrettably, these drugs are not free of side effects. To provide novel treatments for hyperlipidemia, it has been focused on the natural products that have very few side effects [4][5][6].
Herbs have a defined kind of potency through which they can stimulate the human body to protect itself against the diseases. These medicinal herbs serve as a great source of remedies in the treatment of human and animal diseases [7].
Here, we are in progress to prepare a polyherbal formulation which consists of the extract of Allium sativum, Moringa oleifera, Cicer  Garlic (A. sativum L., family: Alliaceae) has played significant medicinal and dietary roles throughout the ages. A. sativum (Garlic) preparations are commercially available in the form of garlic oil and garlic powder, and pills are widely used for certain therapeutic purposes, including improving lipid profile and lowering blood pressure. Extract of A. sativum is alleged to possess valuable effects for the prevention of CVDs. Garlic contains active hypocholesterolemic and hypoglycemic components, known as diallyl disulfide and dipropyl disulfide; it was proven by several studies [8][9][10].
Leaves of M. oleifera Lam., Moringaceae, are claimed to possess several pharmacological activities such as cholesterol-reducing effect and are used to treat patients with heart disease and antiobesity potential that protects the body against adverse effects of high-fat diet-induced obesity. The presence of β-sitosterol in crude extracts of M. oleifera possesses potential hypolipidemic properties [11,12].
C. arientum reported a rich source of vitamins, minerals, and phytoestrogens. Seeds of C. arientum were used as stimulant, aphrodisiac, tonic, anthelminthic, appetizer, relieving burning sensation in stomach, and in the treatment of obesity and in patients who consume excess oily and heavy foods. Cholesterol-lowering effects of C. arientum in different types of hyperlipidemias such as induced by diet are demonstrated in several research studies [13].
H. rosa-sinensis Linn. flowers exhibited a significant reduction in serum lipid parameters such as triglycerides, TC, LDL, VLDL, and increase in HDL [14].
Cholesterol diet and passive smoking raise the lipid and cholesterol levels with reducing the HDL level which causes hypercholesterolemia and hyperlipidemia existing heart disease such as heart attack and heart stroke in the future. Q. indica Linn. raised the HDL level which is good cholesterol and produced a significant reduction in harmful lipids. Q. indica extracts contain flavonoids and phenolic compounds helpful in CVD [15,16].

Authentication of plant material
The plant has been identified and authenticated by Janata PG College, A.P.S. University, Rewa, Madhya Pradesh, voucher specimen no. is Number/J/BOT/H-332 to H-336.
• Soxhlet extraction: Soxhlet apparatus was used for the extraction and hydroalcoholic solvent (1:1) was selected as a solvent for extraction and calculated percentage yield of the extract.     aqueous) before orally administered to the rats. Standard drug was dissolved in suspending agent (1% CMC) before orally administered to the rats [19].
These diets were fed along with normal diet for a total period of 6 weeks to rats.

Experimental protocols
In the HFHSD model, the animals were divided into five groups and each group composed of six animals ( Table 1).
All the treatments were carried out for 42 days. Before and after the treatment, the animals were fasted for 2 h to improve the absorption rate. Parameters studied for this test were body weights, blood glucose, and total high-density lipoprotein cholesterol levels.
• Antihyperlipidemic activity of polyherbal formulation from diet-induced model on experimental rats Acute toxicity studies (LD 50 ): In both Phase I and Phase II procedures, none of the animal mortal or any signs of behavioral changes or show any toxicity on the single administration of Healthcare Administration (2000 mg/kg p.o.). Thus, 300 mg/kg dose was selected for the present study (Table 7).

Evaluation parameters
• Effect on feed (g) and water (ml) intake ( Table 8) • Effect on body weight (g) ( Table 9) • Effect on HDL level (mg/dl) ( Table 10) • Effect on LDL level (mg/dl) ( Table 11) • Effect on CHL level (mg/dl) ( Table 12) • Effect on blood glucose level (mg/dl) ( Table 13) Graphical representation • Effect on total feed and water intake • Effect on body weight • Effect on HDL level

DISCUSSION
The study was carried out to evaluate antihyperlipidemic activity of polyherbal formulation (T1, T2, T3, T4, and T5) in HFHSD-induced model of rats.
In acute oral toxicity study, there were no behavioral changes seen up to 4 h and no mortality was observed up to the end of 24 h even at the maximum tested dose level of 2000 mg/kg per oral. It was considered maximum safe dose.
The study was conducted by measuring various parameters, namely, daily feed intake (g), water intake (ml), body weight (g), lipid profile HDL, LDL, CHL level (mg/dl), and blood glucose level (mg/dl).

CONCLUSION
We can say that polyherbal formulations (T1, T2, T3, T4, and T5) have hypoglycemic activity and significantly improve lipid profile levels in diet-induced experimental rats. Results showed significant decrease in blood glucose level and serum lipid profile such as TC, LDL, and increasing serum HDL level, so could be useful in the treatment of hypolipidemia as mentioned in traditional medicine. However, further studies required to isolate the phytochemicals those responsible for hypolipidemic activity.