Mammary epithelial cell derived exosomal MiR-221 mediates M1 macrophage polarization via SOCS1/STATs to promote inflammatory response.

Lactational mastitis seriously alters the normal physiological function of mammary gland and activates the innate immune. Mammary epithelial cells (MECs) secret cytokines and regulate the function of immune system. However, the mechanism MECs mediated crosstalk with immune cells, such as macrophages, during mastitis is unclear. In this study, mouse mammary epithelial cells (HC11), treated with Lipoteichoic acid (LTA), and macrophages (RAW264.7) were used to mimic intercellular communication. Our results showed that exosomal miR-221 level was up-regulated and reached the peak at 12 h after infected by LTA. The expression of miR-211, CD11b protein and TNF-alpha mRNA were upregulated and the expression of CD206 protein and Arg-1 mRNA were inhibited in RAW264.7 treated with exosomes. In addition, miR-221 mimics and inhibitors enhanced and depressed HC11-derived exosomal miR-221 level, respectively. After treatment of Exo(mimic) in RAW264.7, the expression of CD11b protein and TNF-alpha mRNA were up-regulated, the expression of CD206 and Arg-1 mRNA were down-regulated. Additionally, Exo(inhibitor) enhanced CD206 protein and Arg-1 mRNA levels and inhibited CD11b protein and TNF-alpha mRNA levels. Furthermore, SOCS1 was identified to be a target gene of miR-221 by using Luciferase assays. And western blot assays showed that the expression of p-STAT1 and p-STAT3 were elevated and repressed, respectively. Taken together, we suggest that exosomal miR-221 promotes polarization of M1 macrophages via SOCS1, STAT1 and STAT3. And we reveal a novel crosstalk signaling pathway between mammary epithelial cells and macrophages in the process of inflammation.