The Effects Of Autophagy In Rat On CCl₄-induced Acute Liver Injury And It's Mechanism

Liver is the body's vital organs,with a variety of physiological and biochemical functions,such as the metabolism,the use of amino acids,bile acid synthesis and transformation,and transformation biosynthesis,oxidation protection,detoxification and other physiological functions,which are necessary to maintain the body's healthy functioning.Many science,chemistry,biology and other factors can damage the liver,leading to a series of liver disease,such as liver failure,liver fibrosis and cirrhosis.Autophagy is a lysosomal pathway that degrades and recycles intracellular organelles and proteins to maintain energy homeostasis during times of nutrient deprivation and to remove damaged cell components.Recent studies have identified that autophay plays an important role in the process of liver injury,many functions of the liver-dependent autophagy react.However,for carbon tetrachloride?carbon tetrachloride,CCl₄?in the acute liver injury induced by autophagy and the effect of regulation mechanism is unclear.First of all,to survey the expression level of autophagy in rats' liver regeneration,qRT-PCR,western blotting,immunofluorescence?IF?analysis were used to investigate expression of the autophagy-related proteins,and autolysosomes were investigated by MDC staining in the injury liver induced by CCl₄.After HE staining acute liver injury and liver morphology changes and oil red O staining for fat droplets generated situation.The results indicated that the levels of LC3-? and Atg5 were elevated in 3-6h,then decreased in 12-24h;The levels of SQSTM1/p62 was invreased in 3-6h,then decreased in 12-24 h.LC3,an autophagy-specific protein were detected by IF,the signals were weak and dispersed in normal liver.However,the levels of LC3 proteins became elevated,specific punctate distribution of endogenous LC3-? increased,and MDC fluorescence accumulation spots were also increased after CCl₄ induced liver injury.In summary,autophagy in CCl₄ induced acute liver injury early significantly increased,and in a certain period of time has obvious time-effect manner.To explore the effect of autophagy on liver regeneration,the rats intraperitoneal injection with hydroxychloroquine?CQ,an autophagy inhibitor?for a week co-treatment with CCl₄ induced acute liver injury.We used immunofluorescence analysis to investigate the expression of LC3,MDC staining to exhibit autolysosomes,liver coefficient and HE staining to investigate growth and structure of the remnent liver,Oil red O staining to survey of fat droplets.At the same time take inferior vena venous blood of ALT and AST in serum enzyme activity detection.Immunofluorescence proved that CQ treatment group with a small amount of LC3 protein spots in liver tissue accumulation,CQ joint CCl₄ treatment after 6 h,Furthermore,co-treatment with CCl₄ induced liver injury increased autophagy.CQ treatment group compared with control group,HE staining results found that there is no significant change in shape and structure of the normal liver,however,CQ with CCl₄ group,with the increase of processing time,liver gradually disappear,lobular structure of turbulence;Oil red O staining results showed that the CQ joint CCl₄ treatment group,fat drops significantly increased,the nucleus fragmentation.Enzyme activity test results showed that after CCl₄ hepatic injury of ALT and AST enzyme activity showed a trend of obvious rise,CQ treatment group of enzymes increase degree is more obvious.Finally,immunofluorescence further testing p21 protein expression changes in liver damage.The results show that autophagy is restrained,p21 protein expression level increase?Can be seen from the above results,inhibition of autophagy,the structure and function of the liver and further destruction,damage degree increases,autophagy can be adjusted by suppressing the expression of liver injury p21 protein.In summary the following conclusions,early acute hepatic injury in CCl₄-induced autophagy levels were significantly up-regulated in a time within a certain time-effect manner;the process of acute liver injury induced autophagy in CCl₄-treated play a protective role;autophagy can be adjusted by suppressing the expression of liver injury p21 protein.