Investigations On Pharmacodynamics And Mechanism Of NAFLD Treatment By Huganqingzhi Tablet

Research BackgroundHuganqingzhi tablet is made up of the six traditional Chinese medicines:alisma, notoginseng, cattail pollen, the lotus leaf, hawthorn and pericarpium citri reticulatae which is an empirical prescription from Prof. Kuntang Zang, a famous doctor of traditional Chinese medicine, in Zhujiang Hospital. It could treat the non-alcoholic fatty liver disease for its effects in reduction of liver lipid, improvement of hyperlipidemia and insulin resistance. The pathogenesis network signal of NAFLD was complicated. Silent information factor—SIRT1have taken a vital role in NFLD as reported. SIRT1belongs to the Sirtuin family which was defined as the deacetylation enzyme family that NAD+depended on originally. It could cause the deacetylation of Lysine residues in kinds of proteins. Sirtuin was one Class of highly conserved protein in evolution and involved in many biological events in the cell, especially plays an important role in metabolism, cell survival and life. Many evidences show that NAFLD patients always accompany with obesity, lipid metabolic disorder and insulin resistance. NAFLD have been taken as an essential condition of MS (metabolic syndrome, presented by the American society for clinical endocrinology), but the expression and activity of SIRT1was adjusted by ratio of NAD+/NADH. And the ratio represented the status of substances and energy metabolism in organism or cell. To some extent, SIRT1may play a role of so called nutrient sensors sample, to perceive the substance in the cell energy metabolism through NAD+level fluctuation in cells. So it may play an important role in the regulation of metabolism, the body energy and stable internal nutrition environment. Liver was an important organ in regulation of metabolism, energy and stable internal nutrition environment and blood sugar controlling, and NAFLD was the most common liver disease. So it’s an issue worth concerning that whether the NAFLD was connected with the SIRT1. Huganqingzhi is of great significance in the improvement of NAFLD and expression of SIRT1in the liver organization. NAFLD was a complex pathological syndrome caused by certain liver damaged factors except the alcohol. And it was symbolized by diffused hepatocellular bullae fatty changes, including simple fatty liver and nonalcoholic steatohepatitis or hepatic sclerosis developed from it. Epidemiologic studies showed that NAFLD could influence people with any age or race. In different countries, the ratio of NAFLD patients was about14%-24%, highly in the fat people with ratio of57.5%-74%. NAFLD became the second type of chronic liver disease affected human health and be next only to the hepatitis B. The paroxysm of NAFLD related to Insulin resistance and genetic susceptibility. Metabolic syndrome related fatty liver, such as overweight and the faster pace of increase in weight caused by overnutrition, obesity, diabetes and hyperlipidemia, and cryptogenic fatty liver all belong to the aetiological agents of NAFLD. SIRT1, an important histone deacetylase in human body, play a vital role in regulating body metabolism, stability of energy and nutrients inner environment. Chen suggested that calorie restriction could develop the expression of SIRT1in the high-fat model rats’bodies and improve the NAFLD. Resveratrol as the agonist of SIRT1could improve the insulin resistance of NAFLD rats and regulate hepatic fat. Many indicators showed that SIRT1plays an critical role in the mechanism of NAFLD. Huganqingzhi Tablet consists of six traditional Chinese medicines, alisma, notoginseng, cattail pollen, the lotus leaf, hawthorn, dried tangerine and orange peel. After years of clinical practice, Huganqingzhi Tablet became popular with patients, for its exact curative effect on improving blood biochemical indicators in patients with NAFLD, recovering normal weight and curing fatty liver. But mechanism of how the Huganqingzhi Tablet treat the NAFLD was poorly understood, limited its popularization and application. Therefore, it is important to further clarify the molecular mechanism of action, definite the targets.ObjectiveTo build a model of NAFLD rats by the high fat diet. To explore the treatment effect of Huganqingzhi against the NAFLD. Meanwhile, discuss the mechanism to provide some experimental basis in the basic research and clinical applications of treatment of NAFLD by Huganqingzhi Tablet.MethodAccording to the body weight, randomly divide60♂SD rats into6groups, blank group(lml/kg distilled water), model group(lml/kg distilled water), Fenofibrate group(0.1g/kg), Huganqingzhi high(2.16g/kg)\middle(1.08g/kg)\low(0.54g/kg) dose groups, each group have10rats. Blank group was given common feed, other groups using high fat feed to build model, and give each group rats the corresponding drug in the meantime in12weeks. After model having been built, abdominal aortic blood and kill all the rats. Determine liver related biochemical indexes, blood lipids, blood glucose, liver index, insulin resistance, liver antioxidant ability, etc. Take each rat liver into10%formalin to fasten, serial section and frozen section by normal paraffin, HE staining and oil red O staining. To histomorphology observe the liver lesions situation of each group rats. In the molecular mechanism of experiments, we used the RT-PCR and west-blotting experiment to examine the expression of Silent Information Regulator (SIRT1) in every group rat’s liver.ResultsSuccessful reproducing rat model of nonalcoholic fatty liver disease by12weeks high fat feed. Compared with blank groups, there were satistical differences significantly in the liver protection, liver lipid metabolism, oxidative stress/lipid peroxidation damage and insulin resistance of model group. In Huganqingzhi high and middle dose group, there were certain therapeutic effects in non-alcoholic fatty liverdisease treatment. Huganqingzhi tablet could inhibit aminotransferase elevating and liver enlargement, and has certain protective effect to high-fat diet caused liver injury. Biochemical indicators shows that, Huganqingzhi tablet could obviously improve blood lipid metabolism disorders and liver lipid accumulation in the pathological rats, restrain the elevating of fasting blood glucose and fasting insulin level, increase the insulin sensitive index, reduce insulin resistance index, and improve insulin resistance. Huganqingzhi tablet could reduce blood lipid peroxidation products MDA levels, improve antioxidant enzymes SOD vitality in blood, and also has some protecting effects in oxidative stress/lipid peroxidation injury. Morphology index shows that Huganqingzhi tablet could ease the fatty liver lesions, inflammation necrosis in NAFLD rats. In molecular mechanism experiments, PT-PCR and West blotting experiment shows that compared with blank group, expressions of SIRT1mRNA and protein have been significantly lowered (P<0.05) in model group. But it has been up-regulated(p<0.05, p<0.01) in Huganqingzhi high and middle group.ConclusionsHuganqingzhi tablet could effectively improve insulin resistance, restrain cytokine activation and proliferation of inflammatory mediators, and reduce inflammation response. Furthermore, Huganqingzhi tablet could slowdown oxidative stress/lipid peroxidation injury, restrain the liver collagen deposition, and regulate blood lipid metabolism disorders and inhibiting liver lipid deposition. All in all, in this experiment, we found that Huganqingzhi tablet have intervened occurrence and development of NAFLD in different part and different extent. The possible mechanism that the Huganqingzhi tablet could prevent and cure the NAFLD may via up-regulated the expression of SIRT1mRNA and protein in rats’liver.