Curcumin deactivates M2 macrophages to alleviate lung fibrosis in IgG4-related disease through activating the toll-like receptor 9 pathway

Abstract

Objectives To explore the effects and mechanism of Curcumin on pulmonary fibrosis in IgG4-related disease (IgG4-RD). Methods The expression of fibrosis factors, inflammatory factors and markers of M1/M2 macrophages in lung tissue of IgG4-RD patients was detected by RT-qPCR or Western blotting. The macrophages of IgG4-RD patients were isolated and treated with different concentrations of Curcumin, and the markers of M1/M2 macrophages were detected by RT-qPCR, Western blotting or ELISA. Next, the pulmonary fibroblasts of IgG4-RD patients were isolated and cultured with the supernatant of macrophages treated with Curcumin. Cell proliferation and migration were detected by CCK-8 and Transwell assay, respectively. SOD activity and ROS content were detected by the xanthine oxidase method and flow cytometry, respectively. Then the expression of Toll-like Receptor 9 (TLR9) and its downstream proteins were detected by Western blotting. After treating the cells with TLR9 antagonist IRS869, the changes in the above indicators were further detected. Results The collagen deposition, inflammatory factors secretion and M2 polarization of macrophages were increased in lung tissue of IgG4-RD patients. Curcumin decreased the macrophage M2 polarization and inhibited the proliferation and migration of fibroblasts, reduced the level of oxidative stress, and suppressed the occurrence of fibrosis. The TLR9 pathway was inhibited in IgG4-RD lung tissues, and Curcumin activated this pathway and reduced macrophage M2 polarization. And the inhibitory effect of Curcumin on pulmonary fibrosis could be reversed by IRS869. Conclusions Curcumin inhibited the occurrence of pulmonary fibrosis in IgG4-RD by inhibiting the TLR9 signaling pathway-mediated macrophage M2 polarization.