The Mechanism Of Autophagy In NASH Hepatocyte Mitochondria And The Intervention Effect Of Zhi Gan Prescription

Objective: This subject aims to investigate the NASHmodel rat hepatocyte mitochondrial autophagy mechanism and the intervention effect of Zhi Gan prescription.Methods:80 SD rats are randomly divided into 8 groups: ten 8w former group and ten 12 w former group.One of the former groups will be killed at the end of 8 weeks,and the other will be put to death at the end of 12 th week,both of them were fed with standard diet.The remaining 60 rats were fed with high fat diet: 10 of the them are picked out randomly as 8W Model group and put to death After 8 weeks' high-fat diet.Then the rest rats were randomly divided into five groups: high-dose Zhi Gan Prescription group,low-dose Zhi Gan Prescription group,mid-dose Zhi Gan Prescription group,and Western medicine group and the 12 W model group.The rats were kept fed with high-fat diet at the same time.Make interventions during the 8th week to the 12 th week by give a gavage of Lipitor tablets in western medicine group,and Zhi Gan prescription in high,mid,low dose to the rats in Chinese Medicine group.These 50 rats will be kept fed with high-fat diet until 12 th week.At the end of 12 th week,the blood samples and liver specimens of rats were taken formeasuring the serum levels of AST,ALT,TG and TC by automatic biochemical analyzer.Liver specimens were observed by microscope and ultras-tructure of mitochondrial autophagy was observed by transmission electron micro-scopy.Detecting protein expression levels of PINK1,Parkin,Drp1 and LC3-II through Western Blot test.Results: 1.Two model groups of serum TG,TC,AST,ALT was significantly higher than the same period in the same period of blank group(P < 0.01),and the degree of inflammation of the liver cells and lipid deposition in the same period of blank group is more serious;The hepatic histopathological changes in the 12 W model group were significantly higher than that in the 8W model group,with a large number of hepatic tissue inflammatory responses.It was suggested that the high-fat feeding established the success of NASH,and 8W to 12 W was the progression of NAFLD to NASH.Compared with the 12 W model group,serum AST,ALT,TC and TG levels and pathology in the rats after the intervention were improved in different degrees.It was suggested that both liver and western medicine could improve the liver lesion in NASH rats,and the effect was most obvious in the high-dose group and theremaining treatment group(P < 0.05).2.Observation under the transmission electron microscope: compared with the blank group,model group 12 w hepatocyte mitochondrial morphology change increase,fragmented mitochondria number increased,since the3.decrease in the number of phage,prompt NASH liver cell damage increase in the number of mitochondria and mitochondrial autophagy problems;Each treatment group compared with model group 12 w mitochondria damage degree reducing,fragmented mitochondria decreased,autophagy increases,reducing mitochondrial injury and drug intervention by promoting autophagy was increased liver cell survival rate.The rate of high dose of Zhi Gan prescription group improved significantly.4.Western Blot test showed that the two model group compared with normal group PINK1 protein expression,Parkin,LC3-II are reduced,and elevated Drp1 expression,prompt NASH rat hepatocyte mitochondrial autophagy is restrained,the separatist activities of mitochondria increases;Use after drug treatment,each group rats PINK1,Parkin,LC3-II protein expression and in the same period of the rise of the model group compare to some extent,reduce Drp1 expression,the fat liver side high dose group of each related protein expression changes than the rest of the treatment group is more apparent,prompt Chinese and western medicine treatment can reduce the NASH mitochondrial damage in cells of rats,promote the mitochondrial autophagy,protect liver cell function,reduce the cell death.Conclusion:The NASH rat model can be established by long-term high fat feed feeding.The rats in NASH model had lipid metabolism disorder,blood lipid,liver enzymology and pathological inflammation changes.Damage to mitochondria and inhibition of autophagy may be a key factor in theprogression of the disease.The experimental results showed that the PINK1/Parkin pathway mediated mitochondrial autophagy to participate in the development of NASH.Lipid liver has improved lipid metabolism,liver index and pathology in NASH model rats.This experiment show that the inhibition ofmitochondrial damage and the promotion of mitochondrial autophagy may be one of the mechanisms of Zhi Gan Prescription in treatment of NASH.