Overexpression of miR-378a promotes macrophage M2 polarization and inhibits M1 polarization

doi:10.12307/2024.141

Abstract

Abstract: BACKGROUND: M2 macrophages have the function of reducing inflammatory factors and promoting tissue healing. Therefore, how to regulate M2 polarization of macrophages has been a hot research topic in recent years, and some miRNAs have been found to have this function.
OBJECTIVE: To investigate the effects of miR-378a on the polarization of the Raw264.7 macrophage cell line.
METHODS: The M1 polarization of macrophages was induced by lipopolysaccharide and interferon-γ. Interleukin-4 induced M2 polarization and the expression of endogenous miR-378a in each cell type was detected using qRT-PCR to verify whether miR-378a was involved in the polarization of macrophages. By transfection with lentivirus as the vector of overexpression of miR-378a, the stable expression of miR-378a cell lines was screened. Macrophage M1 polarization was induced synergically by lipopolysaccharide and interferon-γ. Macrophage M2 polarization was induced by interleukin-4. The levels of M1/M2 polarization-related cytokines in the supernatant of the macrophage culture medium were determined by enzyme-linked immunosorbent assay. qRT-PCR was used to detect the polarization characteristics of M1/M2-type macrophages and the mRNA expression levels of related cytokines.
RESULTS AND CONCLUSION: (1) The expression level of endogenous miR-378a in Raw264.7 cells of each group increased after macrophage polarization. (2) Compared with the non-transfected group, the expressions of proinflammatory cytokine-induced nitric oxide synthase, tumor necrosis factor-α, interleukin-6 and interleukin-1β in macrophage M1 induced polarization were significantly decreased in the miR-378a transfection group (P < 0.05); the levels of inducible nitric oxide synthase, tumor necrosis factor-α and interleukin-6 in cell supernatant were also significantly decreased (P < 0.05). (3) Compared with the non-transfected group, the expressions of CD206, interleukin-10 and arginase-I in macrophage M2 induced polarization were significantly increased (P < 0.05); the levels of CD206 and interleukin-10 in cell supernatant were also significantly increased (P < 0.05) in the miR-378a transfection group. (4) It is indicated that overexpression of miR-378a promotes the M2 polarization of macrophages and inhibits the M1 polarization of macrophages.

Key words: miR-378a, macrophage, polarization, lentivirus, inflammatory factor

CLC Number:

Yang Quan, He Huiyu, Wang Sifan, Lyu Shangyi, Zhou Qiqi, Han Xiangzhen. Overexpression of miR-378a promotes macrophage M2 polarization and inhibits M1 polarization[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(13): 2036-2041.

Figures/Tables 8

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