畜牧兽医学报2024，Vol.55Issue(1)：311-322,12.DOI:10.11843/j.issn.0366-6964.2024.01.029
钙结合蛋白S100A4对BCG感染THP-1细胞自噬的调控作用
The Regulation of Calcium-binding Protein S100A4 on Autophagy in THP-1 Cells Infected with Bacillus Calmette-Guérin
摘要
Abstract
The purpose of this study was to explore the regulatory effect of calcium-binding protein S100A4 on autophagy of human monocyte-macrophage THP-1 cells infected with Bacillus Calmette-Guérin(BCG).BCG infected THP-1 cells at different times(MOI=10).The expressions of S100A4 and LC3 proteins were detected by Western blot,and the number of autophagosomes was observed by transmission electron microscopy(TEM).After BCG infection alone or co-treatment with S100A4 small interfering RNA for 12 h,the mRNA expression of S100A4 and autophagy-related factors,LC3 Ⅱ(microtubule-associated protein light chain 3 Ⅱ),Atg7(autophagy-related gene 7),and Beclin1,were detected by qRT-PCR.The protein expressions of S100A4,LC3 Ⅱ,Atg7,and Beclin-1 were detected by Western blot.Autophagy flow was detected by mRFP-GFP-LC3,and spot aggregation of mRFP-LC3 and mRFP-GFP-LC3 was observed by laser confocal microscopy.After BCG was infected with THP-1 cells at different times,the protein expression levels of S100A4 and LC3 Ⅱ firstly increased and then decreased with the extension of infection time,and the expression levels were the highest at 12 h(P＜0.001),and the number of autophagosomes increased significantly.mRNA levels of S100A4,LC3Ⅱ,Atg7,and Beclin-1 in the siNC+BCG infection group were significantly up-regulated compared with those in the siNC group(P＜0.05),S100A4,LC3 Ⅱ,Atg7,and Beclin-1 were significantly expressed at mRNA level after BCG and siS100A4 treatment(P＜0.05)or very significantly(P＜0.01,P＜0.001)down-regulated;Protein levels of S100A4,LC3 Ⅱ,Atg7,and Beclin-1 increased significantly compared with an uninfected group(P＜0.05),the protein expression levels of S100A4,LC3 Ⅱ,Atg7,and Beclin-1 were significantly reduced after BCG and siS100A4 treatment(P＜0.001);Compared with the uninfected group,the number of autophagosomes in BCG group was significantly increased(P＜0.01),the number of autophagosomes in siS 100A4+BCG group was significantly reduced compared with siNC+BCG infection group(P＜0.05).S100A4 has a regulatory effect on the autophagy of macrophages induced by BCG infection,and S100A4 can promote the BCG-induced autophagy of THP-1 cells.关键词
结核分枝杆菌/细胞自噬/S100A4/THP-1细胞Key words
Mycobacterium tuberculosis/autophagy/S100A4/THP-1 cells分类
医药卫生引用本文复制引用
刘悦阳,李梦媛,聂雪伊,马亚博,侯雨欣,马伯利,杨易,徐金瑞..钙结合蛋白S100A4对BCG感染THP-1细胞自噬的调控作用[J].畜牧兽医学报,2024,55(1):311-322,12.基金项目
宁夏自然科学基金项目(2023AAC03201 （）
2023AAC03100) （）
