BCG诱导RAW264.7细胞脂肪酸氧化对细胞自噬和促炎因子表达的调控作用

doi:10.11843/j.issn.0366-6964.2021.09.025

Abstract

Abstract: To investigate the effect of fatty acid oxidation (FAO) on autophagy and the expression of pro-inflammatory cytokines in BCG-infected RAW264.7cells. The accumulation of lipid droplets and the content of fatty acids inside RAW264.7 cells after BCG infection were detected by BODIPY staining and free fatty acid quantitative kit. The expression of carnitine palmityl transferase 1A (CPT-1A) in BCG-infected cells was detected by Western blot. After pretreatment with Etomoxir (100 μmol·L^-1) for 2 h, BCG was infected with RAW264.7 cells for 6 h, intracellular survival BCG was assessed by CFU assay, and the expression of Beclin1, LC3-II and lysosomal protein (RAB7) were detected by Western blot. The aggregation of autophagosomes and autophagy flux were detected by immunofluorescence and mRFP-GFP-LC3 adenovirus. Pro-inflammatory cytokines IL-1β, IL-6 and TNF-α mRNA and their concentration in cells culture supernatant were detected by fluorescence quantitative PCR and ELISA, respectively. The results showed that BCG infection promoted the accumulation of lipid droplets and the expression of CPT-1A in RAW264.7 cells, while free fatty acid content decreased. Etomoxir treatment inhibited intracellular BCG survival and elevated the expressions of Beclin1, LC3-II and RAB7. Besides, a large number of autophagosomes were aggregated and autophagy flux was enhanced, however, the mRNA expressions and secretion of pro-inflammatory cytokines IL-1β, IL-6 and TNF-α were all inhibited. It is suggested that FAO inhibition promotes RAW264.7 cells autophagy and inhibits the inflammatory response induced by BCG infection.

Key words: BCG, fatty acid oxidation, RAW264.7 cells, autophagy, inflammatory

CLC Number:

S855.1

LUO Jia, XU Jinrui, LI Wu, WANG Yujiong. The Effect of BCG-induced RAW264.7 Cells Fatty Acid Oxidation on Autophagy and Pro-inflammatory Cytokines Expression[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(9): 2617-2625.

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The Effect of BCG-induced RAW264.7 Cells Fatty Acid Oxidation on Autophagy and Pro-inflammatory Cytokines Expression

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