| Objective:To investigate the dynamic changes of PD-1, PD-L1, ICOS, ICOSL in the liver and IL-4, IL-2, IFN-Y in the serum in acute liver injury in HBV transgenic mice, in order to study changes mechanism in immunity in HBV transgenic mice with acute liver injury. Methods:20healthy male C57BL/6mice were used as normal control group (group A),20healthy male C57BL/6mice as carbon tetra-chloride (CCL4) group (group B),20male transgenic mice expressing HBsAg negative for transgenic HBsAg negative CCL4group (group C),20male HBsAg positive expression in transgenic mice transgenic HBsAg positive CCL4group (group D), B, C, D were made acute liver injury model, group Awas injected intraperitoneally by0.5ml· kg-1olive oil, sacrificed at Oh,24h,48h,72h to obtain the specimens. B, C, D each group leaving5as control,0.5ml· kg-1intraperitoneal injection of olive oil solution, Oh sacrificed, and15were divided randomly to groups, one-time given and injected by0.5ml· kg-1intraperitoneally10%CCL4olive oil solution, respectively,24h,48h,72h five mice in each group were randomly sacrificed to obtain the specimens, detecting AST, ALT, ALP, TP, ALB, IL-4, IL-2, IFN-γ by serum, while removing the liver, some specimens were used for HE staining and immunohistochemical,others were used for RT-qPCR method of identifying the expression of PD-1, PD-L1, ICOS, ICOSL in the liver.Results:1, Compared group B with group A,24h AST, ALT levels were significantly increased,the difference was statistically significant (P<0.01). group B and D quickly reaches a peak,compared with group C,the difference was statistically significant (P<0.01); at48h, group C peaked, but group B AST has slowly went down, compared with group C and D,the difference was statistically significant (P<0.01).2, Pathology observed:at Oh, group B, C and D, showed no significant pathological changes.24h, group B, C and D were in widespread liver cell degeneration and necrosis mild hepatic lobule may have scattered punctate necrosis, osteoporosis in the cytoplasm and ballooning of the most common;48h, group C disease most severe diffuse of large areas of necrosis;72h, group B, C and D had liver cell cytoplasm and large areas of necrosis and osteoporosis of fat vacuoles, lobular and portal area has a large number of lymphocytes and macrophages dominated inflammatory cell infiltration, but the most severe lesions in C group.3, Immunohistochemistry:group B was highly expressed ICOS and ICOSL at48h, PD-L1at72h; group C was continuing highly expression PD-1from24h to72h, ICOS high expression at24h, PD-L1sustained high expression from48h; group D was highly expressed PD-1, ICOSL at72h.4, RT-qPCR: group B was continuing highly expression ICOS from24h to72h, PD-1expression at24h; group C was continuing highly expression PD-1from24h to72h, ICOS high expression at24h, ICOSL high expression at48h; group D was highly expressed ICOSL at72h.5, CBA: IL-2,group D was highly expressed at Oh,group C was increased significantly at24h,group B and C continued to rise rapidly but decreased in group D at48h, compared D with group B and C, the difference was statistically significant (P<0.01). Group B IL-2peaked at48h,72h down quickly; group C began to increase at24h,48h peaked,72h down quickly. IFN-γ, group B was highly expressed at Oh,group B,C and D were highly expressed at24h, group B and C continued to rise rapidly but decreased in group D at48h, group B, C were reduced at72h,while group D increased, compared D with group B and C,the difference was statistically significant (P<0.01). In the group B IFN-γ began to increase at24h,48h peaked,72h down quickly.The group C Oh,24h,48h and always highly expressed,72h down quickly, group D48h decreased, compared with Oh,24h,72h the difference was statistically significant (P<0.01). IL-4,compared group B with C, D at Oh, the difference was statistically significant.IL-4of expression increased at24h, group B and C continued to rise rapidly but decreased in group D at48h, group B and C decreased rapidly, while group D increased at72h, compared D with group B and C, the difference was statistically significant (P<0.01).In the group B and C IL-4began to increase at24h,48h peaked,72h down quickly. In Group D24h IL-4began to rise,48h minimum,72h rapidly recovered.Conclusion:HBV transgenic mice can occur acute liver injury at24h, especially HBsAg-negative transgenic mice injury was more serious than positive mice. PD-1, PD-L1, ICOS, ICOSL and serum IL-4, IL-2, IFN-γ were increased in normal mouse and transgenic HBsAg negative mouse when were stimulated, and generated strong immune response, but transgenic HBsAg positive mouse had lower expression, may be stimulated by the long-term HBsAg, produced immune tolerance in the body,so the host was in the low level of immune response after CCL4attacked, made host weaker than HBsAg-negative transgenic mice in pathology. |
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