Establishment Of Mouse Model Of Acute Liver Injury Induced By Sodium Cyclamate And Intervention Of Diammonium Glycyrrhizinate Capsule

Objective:The present study is to investigate the intervention effect of diammoniumgly cyrrhizinate enteric-coated capsule(DGEC)in mouse model of acute liver injury induced by intraperitoneal injection of sodium cyclamate.Method: Mouse model of acute liver injury was established by intraperitoneal injection of different doses of sodium cyclamate.The levels of alanine aminotransferase(ALT)and aspartate Amino-transferase(AST)at different time points were determined and liver histology of the mice was examined by Hematoxylin and Eosin staining.Mice were divided into blank group,model group,low-DGEC group,medium-DGEC group and high-DGEC group,and polyene phosphatidyl choline group.Seven days after intraperitoneal injection of sodium cyclamate in all groups but the blank group,mice in model group were gavaged with 0.9% normal saline,those in low-,medium-and high-DGEC groups were treated with respective doses of DGEC,and those in polyene phosphatidyl choline group were treated with polyene phosphatidyl choline for 7 days.On day 14,body weight,liver index,serum levels of ALT and AST,levels of glutathione(GSH),glutathione-peroxidase(GSH-Px),malondialdehyde(MDA)and nitric oxide(NO)in liver tissue were recorded.Then,liver immunohistochemistry was tested by HE staining and ultrastructure of liver cells was observed by electron microscopy.ELISA was performed to determine the expression of tumor necrosis factor-?(TNF-?)in serum and liver.Immunohistochemistry was used to detect the expression of transforming growth factor-?1(TGF-?1)in the liver.Results: 1.Modeling group results: mouse model of acute liver injury was successfully constructed by intraperitoneal injection of sodium cyclamate(4000mg/kg.d)for more than seven days.2.Experimental group results:(1)Liver index,ALT,AST,MDA,and NO levels in sodium cyclamate model group were significantly increased compared with blank group(P < 0.01),while levels of GSH and GSH-Px in model group were significantly decreased compared with blank group(P<0.01).Compared with model group,liver index,ALT,AST,MDA and NO levels of drug intervention groups were all significantly decreased(P < 0.05),while the levels of GSH and GSH-Px in all drug intervention groups were all significantly increased(P < 0.05).Of note,high-DGEC group had the most obvious effect among all drug intervention groups.(2)HE staining showed degeneration of hepatocytes and inflammatory cell infiltration in of the sodium cyclamate model group,while normal states were observed in blank group.In addition,drug intervention groups had different degrees of improvement,and high-DGECgroup had the greatest improvement among all drug intervention groups.(3)Electron microscopy showed decreased mitochondria and endoplasmic reticulum in sodium cyclamate model group,while normal states were observed in blank group.Compared with model group,drug intervention groups had different degrees of improvement.However,the indexes in drug intervention groups failed to return to normal,and differences among drug intervention groups were not significant.(4)ELISA data showed that TNF-? levels in serum and liver of sodium cyclamate model group were significantly increased compared with blank group(P < 0.01).Moreover,drug intervention groups had different degrees of improvement(P < 0.05),and high-DGEC group had the most obvious improvement.(5)Immunohistochemistry showed that TGF-?1 expression in model group was significantly increased compared with blank group(P < 0.05).The levels of TGF-?1 in all drug intervention groups were significantly reduced compared with that of model group(P < 0.05).Of note,there was no difference in the expression of TGF-?1 among drug intervention groups.Conclusion: 1.Intraperitoneal injection of sodium cyclamate can built acute liver injury model in mouse,and present study demonstrates that the occurrence of acute liver injury may be related to oxidative stress,as well as inflammatory reaction mediated by TNF-? and TGF-?1.2.DGEC has therapeutic effect on mouse model of acute liver injury induced by sodium cyclamate.