| Non-alcoholic fatty liver disease(NAFLD)is a kind of hepatic steatosis characterized by significant lipid deposition in the hepatocytes(steatosis)after the exclusion of significant alcohol intake,viral infection,or other specific liver disease,which manifested as a wide histological spectrum of liver damage,ranging from isolated steatosis to non-alcoholic steatohepatitis.As a serious public health issue,this disease encompasses a spectrum of changes ranging from steatosis and steatohepatitis to fibrosis and hepatic cirrhosis,and is associated with a higher frequency of hepatocellular carcinoma.The precise pathogenetic mechanisms underlying the development and progression of NALFD have not been completely clarified yet.Current therapies for fatty liver disease aimed at reducing body weight and improving insulin sensitivity to alleviate the associated metabolic syndromes are not satisfied.Traditional Chinese medicine(TCM)is commonly used in treating liver diseases worldwide,especially in China.Combinatory therapeutic strategies according to patient symptoms are often applied to various types of chronic diseases.The effects of traditional Chinese herbs on metabolic parameters associated with NAFLD have been studied in vitro and in vivo.Hence,in the current study,we aimed to examine the protecting effect of Hua Tan Qu Shi(HTQS)decoction decoction to liver injury during NAFLD.ObjectivesIn this study,the animal model of SD rats infected with high-fat-diet-induced hyperlipidemia and nonalcoholic fatty liver disease was used to test blood-related indicators and conduct liver histopathological and morphological exploration,and thereby observe the effects of Hua Tan Qu Shi decoction on lipid metabolism in the case of high Hyperlipidemia and nonalcoholic fatty liver disease.Through investigating molecular pathways in autophagy and apoptosis of hepatic cells of SD rats,this study aims to explore the possible mechanism in which Hua Tan Qu Shi decoction improves injury induced by nonalcoholic fatty liver disease.Methods6-week-old male SD rats were fed with high-fat diet for 9 weeks.50 grownup rats,according to their weight and blood fat,were randomly divided into control group,L-HTQS group,M-HTQS group,and Atorvastatin group(10 rats in each group),being fed with original high-fat diet for 10 weeks.The control group was given the same volume of pure water.10 SD rats were used as a normal control group fed with basal diet and equal volume of pure water every day.These rats were weighed at the same time each week,and their reactivity,hair color,diet,eyes,excreta and mental state were closely observed throughout the course of the experiment.After the experiment,the liver was weighed and the liverbody weight ratio was calculated.The levels of TG,TC,HDL-C,LDL-C and FFA in the blood taken from aorta abdominalis were measured;the serum ALT and AST were also measured.Changes of serum liver function related indicators ALT,AST,ALP,endothelial function related indicators NO,NOS,ET-1 were tested.HE and Oil Red O staining were used to observe the lipid accumulation and histomorphological changes in hepatocytes.Immunohistochemistry was used to detect the expression of LC3-b and Caspase-3 in paraffin sections of liver tissue.The mRNA expression of autophagy and apoptosis-related genes Beclin-1,Caspase3,LC3-b,SREBP and Bcl-2 in liver tissue were detected by RT-PCR.And lastly,the method of western blot was used to detect the protein expression of P62,Caspase-3,LC3-b,SREBP and Bcl-2 in the liver.ResultsBody weight and liver weight:the body weight of rats in each group increased continuously,and the body weight and liver-body weight ratio of SD rats were significantly increased by high fat diet.The weight gain of rats in the treatment group was delayed,but there was no significant difference.The high dose of phlegm-dampness prescription significantly reduced the rat liver weight ratio.Lipid Metabolic Index:in terms of serum lipid,TG and FFA(P<0.05)were significantly decreased in the group treated with high dose of phlegm-dampness prescription,which also showed a decreasing trend in TC,LDL-C.In contrast,groups with medium and low doses failed to show significant changes.Liver pathological changes:groups with high dose of phlegm-dampness prescription significantly improved liver cell steatosis,reduced deposition of lipid droplets in the liver and inflammatory cells in the liver infiltration,and maintained normal structure of liver cells.Liver function:Medium and high dose of phlegm-dampness prescription could significantly decrease the levels of AST,ALT and ALP(P<0.05).There was no significant change in the low dose group(P>0.05).Endothelial function:The levels of NO,NOS were significantly increased while that of ET-1 decreased(P<0.05)in the high and low dose group.There was no significant change in the medium dose group.Immunohistochemistry:In high dose group,the leve of LC3-b increased and the expression of Caspase-3 was inhibited.There was no such obvious effects observed in the low dose group.Western blotting and mRNA expression:The expressions of Caspase-3,SREBP and P62 mRNA were significantly inhibited(P<0.05)and those of Bcl-2 and LC3b mRNA increased in the high dose group.No similar effects were found in the medium and low dose groups.In terms of protein expression,the expressions of Caspase-3,SREBP and P62 in the liver tissue were significantly down-regulated in the high dose group compared with the other two groups(P<0.05).Conclusions(1)Hua Tan Qu Shi decoction can reduce the blood lipid levels of SD rats infected with high fat diet induced hyperlipidemia combined with nonalcoholic fatty liver disease.The effect of high dose treatment is better.(2)Hua Tan Qu Shi decoction has a protective effect on liver injury in rats with nonalcoholic fatty liver disease.The effect of high and medium dose treatments is relatively better.(3)Hua Tan Qu Shi decoction has positive functions in the overall mediation of oxidative stress of the experimented rats,improves the body’s antioxidant capacity,and protects the endothelial function.The effect of high dose is the best,followed by the medium dose.(4)Hua Tan Qu Shi decoction can inhibit the expression of apoptosis-related factors and activate the key factors of autophagy pathway,thus preventing liver injury and reducing the accumulation of lipid in hepatocytes.In this regard,the effect of high dose is the best.In this study,non-alcoholic fatty liver disease animal model of SD rats induced by high-fat diet was used to simulate the pathogenesis of human disease.On the basis of this,this study uses the high fat induced animal model to explore the effects of the Hua Tan Qu Shi decoction on the improvement of non-alcoholic fatty liver disease.This is the first time that the Hua Tan Qu Shi decoction and studies on autophagy and apoptosis related pathways are integrated to understand the mechanism of its improvement effects on injury caused by nonalcoholic fatty liver disease from the perspective of autophagy and apoptosis. |
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