Effect Of Sinomenine And Bone Marrow Mesenchymal Stem Cells On Immune-induced Liver Injury Induced By Con A In Rats

ObjectivesThis study investigated whether the combination of Sinomenine and bone marrow mesenchymal stem cells can treat Con A-induced immune liver injury model,and tried to elaborate its possible mechanism,and provide a theoretical basis for the application of Sinomenine + BMSCs in the treatment of immune hepatitis.Methods1.Con A induced acute immune hepatic injury in rat model Forty-two Wistar rats were randomly divided into 6 groups.N group was normal control group.A,B,C,D and E were injected with 4mg/kg、8mg/kg、16mg/kg、30mg/kg and 40mg/kg Con A respectively at the 8th hour.The levels of ALT、AST、ALB,IL-2 and IL-10、IFN-γ、TNF-α were detected,And HE staining was used to calculate the inflammatory activity,and the number of abnormal deaths was calculated at the end of each experiment;Twenty-one Wistar rats were randomly divided into normal control group,intraperitoneal injection group and tail vein injection group,with 7 rats in each group。Injecte 16mg/kg concanavalin A in rats through the tail vein and abdominal cavity,comparing serum ALT、AST、ALB respectively.2.Sinomenine protects Acute Hepatic Injury Induced by Con A by Immunomodulation Fourty-nine female Wistar rats were randomly divided into7 groups: model group,normal group,SIN group(A 60mg/kg,B 90mg/kg,C 120mg/kg,D 150mg/kg)、Dex,with 7 rats in each group.16 mg/kg Con A intravenously establish a liver injury model,except for the normal group.Dex group,SIN different dose group were administered 1h before Con A injection,SIN group injected one more time after 3h.After 8h injection of Con A,blood was taken from the abdominal aorta and liver tissues were harvested.The levels of ALT、AST and ALB were detected by automatic biochemical analyzer.The levels of IL-2、IL-10 were detected by Elisa.3.Effect of Sinomenine and bone marrow mesenchymal stem cells on immune-induced liver injury induced by Con A in rats Forty-two female Wistar rats were randomly divided into 6 groups: normal group,model group,SINomenine,mesenchymal stem cell group,SINomenine + mesenchymal stem cell group,dexamethasone sodium phosphate group,dexamethasone phosphate Sodium and mesenchymal stem cells were administered 1 h prior to injection of ConA into the caudal vein.SIN administration was performed as before.After 8h injection of Con A,blood was taken from the abdominal aorta and liver tissues were harvested.The levels of ALT,AST,ALB,TBIL,IL-2,IL-10,IL-10 and IFN-γ were detected and the ratio of CD4 / CD8 was detected by flow cytometry.Hematoxylin.Results1.the number of abnormal deaths at the end of each experiment: 0 in group A、B、C 2 in group D and 7 in group E.HE staining showed.A,B group showed a small amount of punctate necrosis,inflammatory cell infiltration,obvious bridging-like necrosis in C and D groups,leaflet structure is basically complete;Serum ALT,AST,ALB is no obvious abnormalities,normal group and intraperitoneal injection group2.Compared with the model group,the levels of ALT,AST and IL-2 in Dex group and SIN group were significantly decreased,while the levels of ALB and IL-10 in the model group were significantly increased(P<0.05),and histopathological analysis showed The levels of ALT,AST and IL-2 in serum of SIN 60mg/kg group and SIN 90mg/kg group were significantly lower than those of SIN 120mg/kg group,while the levels of ALB and IL-10 increased significantly(P <0.05).There was no significant difference in ALT,AST,ALB,IL-2 and IL-10 levels between SIN 120mg/kg and SIN 150mg/kg group(P> 0.05).The Dex group and the SIN 120mg/kg group were compared,each index difference was statistically significant(P<0.05).3.The third generation of rat BMSCs were subcultured and their cells were long fusiform with the typical morphology of MSCs.Compared with SIN group and BMSCs group,the levels of ALT,AST,TBIL,IFN-γ and CD4 + / CD8 in SIN+ BMSCs group decreased 0.05),IL-10,IL-4 increased.Conclusion1.Rat model of acute liver injury was successfully induced by injection of 16 mg / kg concanavalin A.2.SIN can protect Con A-induced acute immunity-induced liver injury and inhibit the activation of T lymphocytes and regulate the secretion of IL-2and IL-10,which may be one of its mechanisms.3.SINomenine and mesenchymal stem cells combined with immune synergies.Possible mechanisms are to suppress the secretion of Th1 cytokines,modulate the Th1 / Th2 ratio,and predispose the immune status to suppress cellular immunity.