Adipophilin Induces The Expression Of Inflammatory Cytokines Through ERK1/2-AP-1Signal Pathway

Objective: To observe the effects of adipose differentiation-related protein (adipophilin,ADRP) on the expression of inflammation factor in RAW264.7mice macrophage and theunderlying mechanisms.Methods: Constructed high expression and low expression adipophilin RAW264.7cellsmodels by transfecting PA317packaging cells with high and low stable adipophilinretroviral vector into RAW264.7cells separately, RT-PCR and Western blot methodswere used to detect the expressions of IL-6、MCP-1、TNF-α mRNA and protein levels inRAW264.7cells. ELISA method detected the expressions of IL-6, MCP-1, TNF-α inmedium. Western blot detected the expressions of AP-1, phosphorylated AP-1, ERK1/2and phosphorylated ERK1/2. Treated RAW264.7cells models mentioned above with theERK1/2blocker, PD98059, and AP-1blocker, curcumin, separately, and used Westernblot and ELISA methods to test the expressions of adipophilin、p-ERK1/2and p-AP-1proteien and inflammation factors.Results: Comparing with RAW264.7cells in the normal state, RAW264.7cells with highexpression of adipophilin had higher level of IL-6、MCP-1and TNF-α, and up-regulatedthe expression of phosphorylated AP-1and phosphorylated ERK1/2, while theRAW264.7cells with low expression of adipophilin had opposite results. ERK1/2inhibitor had no significant effect on the expression of adipophilin (P <0.05), but theexpression of ERK1/2protein and AP-1protein were significantly inhibited (P <0.05).The administration of AP-1inhibitor, curcumin, had no significant effect on theexpressions of adipophilin protein and ERK1/2protein, but the expression of AP-1 protein was significantly inhibited (P<0.05). At the same time, the expression ofinflammatory cytokines IL-6、MCP-1and TNF-α were significantly lowered (P <0.5)Conclusion: Adipophilin may regulate the expression of inflammatory factors throughERK1/2-AP-1pathway in RAW264.7macrophages.