Study On The Preventive Effect And Mechanism Of Jincao Disinfection Granule On DEN-Induced Hepatocellular Carcinoma In Rats

Objective:Jincao Xiaodu Granules(JCG)is an experienced prescription of Guangxi Zhuang Medicine.Previous studies have proved that it has significant effects on acute liver injury and liver cancer.The purpose of this study was to investigate the therapeutic mechanism of Jincao Xiaodu Granules on rats hepatocellular carcinoma(HCC)induced by diethylnitrosamine(DEN),a potent chemical carcinogen.Method:All experiments were carried out in rats where DEN-induced hepatocellular carcinoma model was constructed.The rats were randomly divided into 5 groups:control group,DEN group,and JCG-L,M,H groups(1700,3400,and 6800 mg/kg).The physiological state of rats in each group was observed after 6 week-continuous administration.After the last dose,the rats could not fast from water for 12-16 hours.Blood samples were collected from the abdominal aorta and liver samples were achieved after the rats anesthetized with chloral hydrate.The hepatocyte structure and pathological changes were observed by hematoxylin-eosin(HE)staining.The liver function indicators of ALT,AST and TBIL were detected by automatic biochemical analyzer.Serum alpha fetoprotein(AFP)and transforming growth factor beta 1(TGF-?1)were determined by ELISA.The activity of superoxide dismutase(SOD)was determined by WST-1 method,and malondialdehyde(MDA)was determined by TBA method.Expression of programmed death receptor-1(PD-1)and programmed death ligand-1(PD-L1)in liver tissues was detected by Western-blot.The expressions of PD-1,PD-L1 and TGF-?1 were detected by immunohistochemistry(IHC-P)of paraffin sections.Results:Compared with the control group,some cells in the liver pathological sections of rats in the DEN group were lysed and transformed into vacuoles.Increasing number of heteromorphic cells and basophilic nuclei and enlarged volume of nucleoli could also be observed.In the meantime,pathological mitosis had existed and ALT,AST,and TBIL levels were significantly increased(P<0.01).SOD activity decreased significantly(P<0.01),and MDA content increased significantly(P<0.01).The expressions of PD-1,PD-L1,AFP and TGF-?1 were up-regulated and statistically significant(P<0.01).Compared with DEN group,ALT,AST and TBIL levels were significantly decreased in the three doses of JCG groups(P<0.01).According to HE staining,JCG significantly improved DEN-induced Histopathological status of rat HCC.The expressions of PD-1,PD-L1,AFP and TGF-?1 were down-regulated.Among them,JCG-H group played the most significant role.Conclusion:Experimental studies have shown that JCG can effectively improve DEN-induced HCC in rats,and plays a therapeutic role by blocking the immune escape pathway of PD-1/PD-L1 to reduce oxidative damage.It may be related to the improvement of liver function,the promotion of liver tissue self-healing and the enhancement of the body's antioxidant capacity.