Background: The mechanisms of macrophages/monocytes in autoimmune hepatitis (AIH) remain unclear. We investigated the role of receptor-interacting protein kinase 3 (RIP3), a key inflammatory signal adapter, in macrophage/monocyte activation in AIH. Methods: Liver tissues and monocytes from patients were collected to evaluate the relationship between macrophage activation and RIP3 by double-immunofluorescence and Western blotting. RAW264.7 macrophages were used to study the regulation of RIP3 signaling on inflammatory cytokines. Results: Compared to the hepatic cyst, the majority of accumulated macrophages expressed RIP3 in AIH liver tissues. Moreover, RIP3 expression of monocytes was correlated with the levels of serum hepatic enzyme in AIH. Furthermore, RIP3 signaling was activated by lipopolysaccharide in RAW264.7 macrophages, which was accompanied with upregulated interleukin (IL)-1b, IL-6, and IL-10 and downregulated IL-4 and transforming growth factor-b. Notably, necrostatin-1, the specific inhibitor of the RIP3 signaling pathway, and 6-thioguanine (6-TG), the active metabolite of azathioprine, predominantly reduced IL-6 production compared to other cytokines. Moreover, the gene level of IL-6 was dramatically increased in AIH liver tissues. Conclusions: RIP3 signaling is involved in macrophage/monocyte activation in AIH and mediates IL-6 production, and is a novel molecular mechanism of 6-TG, indicating that it might be a promising therapeutic target for AIH treatment.
KeywordsAutoimmune hepatitis, macrophages, receptor-interacting protein kinase 3, interleukin-6Received: 4 October 2017; accepted: 2 January 2018
Key summaryOur study determined a novel molecular mechanism of macrophage/monocyte activation and inflammatory response in autoimmune hepatitis (AIH). Moreover, the role of receptor-interacting protein kinase 3 (RIP3) in human AIH disease was reported for the first time. In addition, this study points out the relationship between interleukin (IL)-6 and AIH, and IL-6 can be downregulated through inhibiting the RIP3 signaling of macrophages. Finally, our investigation provided a novel mechanism for the effect of thiopurines on the immune system.