脂肪分化相关蛋白2对BCG诱导小鼠传代巨噬细胞自噬的调控作用

doi:10.11843/j.issn.0366-6964.2023.05.034

Abstract

Abstract: The aim of this study was to investigate the role of perilipin 2(PLIN2) in the regulation of autophagy in BCG infected mouse macrophages RAW264.7. In this study, we used small interfering RNA technology to knock down the expression of PLIN2 in murine macrophages RAW264.7, combined with BCG infection, and detected intracellular PLIN2 protein expression changes and indicators of autophagy, endoplasmic reticulum stress and fatty acid metabolism-related factors by immunoblotting, flow cytometry and immunofluorescence. BCG infection significantly upregulated the expression of PLIN2 (P<0.05) and highly significantly upregulated the expression of autophagy-associated proteins ATG5 (P<0.01), ATG12 (P<0.001) and LC3 (P<0.001) in macrophages RAW264.7 and was accompanied by intracellular neutral lipid accumulation. Knockdown of PLIN2 significantly downregulated autophagy-related proteins ATG5 (P<0.05) and ATG12 (P<0.05) and LC3 (P<0.001) in BCG-infected macrophages RAW264.7, significantly reduced autophagy rate (P<0.001) and significantly decreased intracellular neutral lipid content (P<0.001). At the same time, knockdown of PLIN2 significantly downregulated CHOP (P<0.05) and highly significantly downregulated ATF4 (P<0.001) endoplasmic reticulum stress-related protein expression, and intracellular Ca²⁺ concentration was highly significantly reduced (P<0.001). Finally, we treated cells with the endoplasmic reticulum stress agonist Tunicamycin and found no significant difference in intracellular autophagy-related protein. BCG infection of mouse macrophages RAW264.7 promoted the expression of PLIN2 and increased the accumulation of neutral lipids, which in turn activated the PERK-eIF2α-ATF4-CHOP signaling pathway in the endoplasmic reticulum stress pathway and induced the onset of cellular autophagy.

Key words: PLIN2, BCG, macrophage RAW264.7, endoplasmic reticulum stress, cellular autophagy

CLC Number:

WANG Chongnian, YU Jialin, GONG Zhaoqian, WU Xiaoling, DENG Guangcun. Regulation of BCG-induced Autophagy in Macrophages RAW264.7 by PLIN2[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2134-2146.

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Regulation of BCG-induced Autophagy in Macrophages RAW264.7 by PLIN2

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