To Study The Protective Effect Of Erzhi Pills On Actue Liver Injury Caused By Endoplasmic Reticulum Stress Induced By CCl₄ In Mice

ObjectiveThe aim of the study was to establish the model of carbon tetrachloride?CCl₄?induced endoplasmic reticulum stress resulted in acute liver injury in mice and to study the protective effect of Erzhi pills.The study would provide the theory basis of Erzhi pills for clinical prevention and treatment of liver diseases.Methods:1.The preparation of Erzhi pills:weighting the eclipta and processed fructus ligustri lucidi powder amount in advance in a cooking pot,adding water soaking,decoction,filtering,collecting filtrate,then add a small amount of water,decoction,filtering,collecting filtrate,merging filtrate,moderating concentration.According to Chinese pharmacopoeia test method for quality evaluation.2.50 kunming mice were randomly divided into normal group and 4 groups of CCl4 model groups.The acute liver injury murine model was induced by intraperitoneal injection of 10%CCl₄,while the normal control group received only physiological saline as control.Plasma levels,such as aspartate aminotransferase?AST?,alanine aminotransferase?ALT?,superoxide dismutase?SOD?,glutathione peroxidase?GSH-Px?and malondialdehyde?MDA?,were measured at 16,24,48 and72 h after injecting.Calculating the lover index and spleen index.Histological examination was performed using HE staining and apoptosis was detected using TUNEL.The ERS related proteins,CHOP and GRP78,were detected by immunohistochemistry method.3.A total of 50 mice were randomly divided equally into control group,model group and the low,medium and high dose of Erzhi pills groups?140?280?560mg/ml?.The mice in the drug treatment groups were administered different doses of Erzhi pills via intragastric gavage?0.1ml/10g?once per day for 14 days,while those in the normal control group and model group were administered equal volume of physiological saline.8 h after the last administration,each group of mice was injected with CCl₄?0.1ml/10g?by intraperitoneal injection except for the normal control group.The mice were killed 24 days later and then calculated the lover index and spleen index.The concentration of the serum aspartate aminotransferase?AST?,alanine aminotransferase?ALT?,superoxide dismutase?SOD?,glutathione peroxidase?GSH-Px?and malondialdehyde?MDA?were measured by extracting the eyeball blood.Histological examination was detected by hematoxylin-eosin?HE?staining and apoptosis was detected by TUNEL.The ERS related proteins,CHOP and GRP78,were detected by immunohistochemistry method and Western Blot.Results:1.According to Chinese pharmacopoeia method for the test of Erzhi pills,the results conformed to the standard.2.Compared with normal group,the levels of ALT,AST and MDA of model groups were all increased,and reached the peak at 16 h?P<0.01?,then gradually decreased to normal level.The activities of SOD and GSH-Px in model groups decreased significantly?P<0.01?in 24 h,then increased;the liver index of model groups were significantly increased?P<0.01?in 24h and then decreased,and the spleen index was contrary to it.The results of HE staining showed that the hepatocyte of model groups were swelling,cytoplasm rarefaction,spotty necrosis and ballooning degeneration.The results of TUNEL showed that the apoptosis index of model groups increased significantly?P<0.01?.Immunohistochemical results showed that the expression of CHOP and GRP78 significantly increased?P<0.01?compared with normal group,and reached the peak at 24 hours.3.Compared with model group,the serological indexes in different doses of Erzhi pills groups were significantly improved?P<0.05 or P<0.01?,the extent of liver lesion was decreased and the degree of injury was significantly reduced,and the liver index and spleen index of mice were significantly changed?P<0.05 or P<0.01?.The results of TUNEL showed that the apoptosis index in different doses of Erzhi pills groups decreased significantly?P<0.01?,and the expression of CCAAT/enhancer binding protein homologous protein?CHOP??glucose regulated protein 78?GRP78?were significantly decreased compared with model group?P<0.05 or P<0.01?.Conclusion:It was the most obvious that the acute liver injury in 24 h induced by ERS after treated with 0.1 ml/10 g 10%CCl₄ by intraperitoneal injection.Erzhi pills has a significant protective effect on ERS induced acute liver injury,and the mechanism may be involved in the inhibition of ERS and down-regulation of ERS marker apoptosis protein CHOP expression.