Regulatory Effect Of CQMUH-011 On Immune Function

Objective To investigate the effect and mechanism of CQMUH-011on the immune system in mice.Methods Freshly isolated mouse splenocytes were stimulated with lipopolysaccharide?LPS?or Concanavalin A?Con A?,effects of CQMUH-011 on the proliferation of T and B lymphocytes were assessed with CCK-8 assay.Carbon clearance test and peritoneal macrophage phagocytosis test were carried out to detect the effect of the drug on non-specific immune function;and the influence of the drug on the ability of antibody production was explored by the serum agglutination experiment;moreover,in order to test the effects of the drug on cellular immunity and immune organ index,a mouse model of delayed-type hypersensitivity?DTH?induced by dinitrofluorobenzene?DNFB?was established.In addition,Con A?20mg/kg,iv?was injected into mice to induce autoimmune hepatitis?AIH?through the tail vein.Hematoxylin and eosin?H&E?staining was used to evaluate the histopathology pathological changes of liver and spleen;at the same time,enzyme-linked immunosorbent assay?ELISA?was applied to measure the serum levels of inflammatory factors;and the levels of serum transaminase,malondialdehyde?MDA?and myeloperoxidase?MPO?were detected by appropriated kits,respectively.Inflammation-related proteins that expressioned in liver tissue were checked by western blot.Then,flow cytometry?FCM?was used to investigate the subsets of splenic T lymphocytes and apoptosis of thymocytes,and the correlation between serum alanine aminotransferase?ALT?levels and lymphocyte subsets were also analyzed.Results?1?The new compound CQMUH-011 inhibited the proliferation of splenic T and B lymphocytes;it also decreased the rate of carbon clearance and the phagocytosis of peritoneal macrophages,as well as the production of serum antibodies.Meanwhile,in DTH model,it could significantly ameliorate the mouse ear swelling degree and reduce the glandular index of spleen and thymus.?2?The new compound CQMUH-011 could significantly elevated the ratio of CD3⁺CD4⁺/CD3⁺CD8⁺and lowered the porporation of CD4⁺CD25^-CD69⁺in splenocytes of AIH mice at the same time,and inhibited the activation of T lymphocytes,apoptosis of thymocytes and secretion of cytokines,thus reducing hepatic pathological damage in AIH mice,decreasing the levels of ALT and AST and the expression of NF-?B as well as its upstream related proteins,and also reduced the production of MDA,MPO.?3?Serum ALT levels were correlated with CD4⁺CD25^+/-CD69⁺lymphocyte subsets,especially CD4⁺CD25^-CD69⁺in AIH mice model.Conclusion These results demonstrated that CQMUH-011 has a moderating effect on immune function and protects mice from Con A-induced AIH.