HIF-1α/BNIP3信号通路对BCG诱导巨噬细胞自噬的影响  

Role of HIF-1α/BNIP3 signaling pathway in regulating macrophage autophagy induced by BCG infection

作　　者：牛莎莎 于志瑞 邓光存[1,2] 吴晓玲[1,2] NIU Shasha;YU Zhirui;DENG Guangcun;WU Xiaoling(Key Laboratory of Ministry of Education for Conservation and Utilization of Special Biological Resources in the Western China,Yinchuan 750021,Ningxia,China;School of Life Sciences,Ningxia University,Yinchuan 750021,Ningxia,China)

机构地区：[1]西部特色生物资源保护与利用教育部重点实验室,宁夏银川750021 [2]宁夏大学生命科学学院,宁夏银川750021

出　　处：《微生物学通报》2022年第9期3837-3848,共12页Microbiology China

基　　金：国家自然科学基金(32060160,32160162);宁夏回族自治区重点研发计划(2018BFH03017)。

摘　　要：【背景】缺氧诱导因子1α(hypoxia-inducible factor 1-alpha,HIF-1α)是响应细胞低氧反应的关键因子,在红细胞生成、血管形成、能量代谢及调节宿主免疫代谢中发挥着重要作用。【目的】探讨HIF-1α/Bcl-2-腺病毒E1B相互作用蛋白3(Bcl-2-adenovirus E1B 19-kDa interacting protein 3,BNIP3)信号通路对牛分枝杆菌卡介苗(Bacillus Calmette-Guérin,BCG)诱导小鼠巨噬细胞RAW 264.7自噬的影响。【方法】构建HIF-1α的小干扰RNA(siHIF-1α),转染RAW 264.7细胞后,结合BCG感染,采用流式细胞仪检测细胞自噬率,用Western blotting或免疫荧光技术检测HIF-1α、BNIP3、LC3、Beclin 1、Rheb和mTOR的表达水平。【结果】BCG感染显著上调巨噬细胞中LC3和HIF-1α的表达,用siHIF-1α结合BCG感染后显著下调巨噬细胞中HIF-1α、BNIP3、LC3、Beclin 1和细胞自噬率水平,并促进Rheb和p-mTOR的表达。【结论】在BCG感染RAW 264.7细胞过程中,干扰HIF-1α表达抑制了HIF-1α/BNIP3信号通路,进而激活了mTOR途径,抑制BCG感染诱导的细胞自噬。[Background]Hypoxia-inducible factor 1-alpha(HIF-1α)is a key factor in response to cellular hypoxia and plays an important role in erythropoiesis,angiogenesis,energy metabolism and regulation of host immune metabolism.[Objective]To investigate the effects of HIF-1α/Bcl-2-adenovirus E1B 19-kDa interacting protein 3(BNIP3)signaling pathway on the BCG infection-induced autophagy of macrophage RAW 264.7 cells.[Methods]Small interfering RNA of HIF-1α,siHIF-1α,was constructed,and after the transfection of siHIF-1αand/or Bacillus Calmette-Guerin(BCG)infection of RAW 264.7 cells,the autophagy rate of the cells was detected by flow cytometer.Western blotting or immunofluorescence technique was employed to determine the protein levels of HIF-1α,BNIP3,LC3,Beclin 1,Rheb and mTOR.[Results]BCG infection up-regulated the expression of LC3 and HIF-1αin RAW 264.7 cells.The transfection of siHIF-1αdown-regulated the levels of HIF-1α,BNIP3,LC3,and Beclin 1 and decreased the autophagy rate of the macrophages after BCG infection.Moreover,siHIF-1αpromoted the expression of Rheb and p-mTOR.[Conclusion]Our results indicated that the knockdown of HIF-1αinhibited the HIF-1α/BNIP3 signaling pathway,thereby activating the mTOR pathway and inhibiting autophagy in RAW 264.7 cells after BCG infection.

关 键 词：缺氧诱导因子1Α Bcl-2-腺病毒E1B相互作用蛋白3 牛分枝杆菌卡介苗 巨噬细胞RAW 264.7 自噬 

分 类 号：R392[医药卫生—免疫学]