鲍曼不动杆菌OmpA经ROS/NLRP3信号通路调控THP1细胞自噬和凋亡

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Abstract Objective: To investigate the effect of outer membrance protein A(OmpA) of Acinetobacter baumannii on autophagy and apoptosis of human monocyte macrophage THP-1 cells and its underlying mechanism. Methods: The OmpA/pET-28a vector was constructed and transformed into E.coli BL21(DE3) to express the recombinant OmpA (rOmpA), and the rabbits were immunized by rOmpA to prepare the polyclonal anti-OmpA antibody. The anti-OmpA antibody levels in serum of ventilatorassociated pneumonia(VAP) patients were detected by Western blotting. THP-1 cells were treated with different concentrations(1 μg/mL and 10 μg/mL) of rOmpA for 1 h, 3 h and 6 h; and the expression of autophagy-related proteins LC-3 and Beclin-1 were detected by Western blotting; the flow cytometry was used to detect the THP-1 cell apoptosis rate and ROS production; the mRNA expressions of NLRP3, Caspase-1 and IL-1β in THP-1 cells were detected by qRT-PCR. Results: The OmpA expression in clinical A. baumannii strains could be detected by polyclonal anti-OmpA antibody, and the anti-OmpA antibody (IgG) in serum of VAP patients serum could be detected before and after the onset of VAP. The levels of anti-OmpA antibody in VAP patients′ serum were significantly higher than that before using tracheal intubation (P<0.05). After treated with rOmpA, the expression of autophagy-related proteins LC3-Ⅱ and Beclin-1 in THP-1 cells increased significantly, and the apoptosis rate increased vastly in a time- and concentration-dependent manner (P<0.05). Compared with the control group, when cells were treated by the rOmpA for 1 h, the IL-1β mRNA expression and reactive oxygen species were significantly up-regulated (P<0.05); when treated for 3 h and 6 h, the three kinds of mRNA expression had a significant decrease compared with the control group (P<0.05). Conclusion: OmpA induces THP-1 cells autophagy and apoptosis by activating NLRP3 inflammatory cytokines and releasing inflammatory factors such as ROS and IL-1β.

Received: 12 January 2020

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http://zzs.ujs.edu.cn/xbyxb/EN/ OR http://zzs.ujs.edu.cn/xbyxb/EN/Y2020/V30/I05/423

［1］Xie J, Yang Y, Huang Y, et al. The current epidemiological landscape of ventilatorassociated pneumonia in the intensive care unit: a multicenter prospective observational study in China\[J\]. Clin Infect Dis, 2018, 67(suppl_2):S153-S161.

［2］Li FJ, Starrs L, Burgio G. Tug of war between Acinetobacter baumannii and host immune responses\[J\]. Pathog Dis, 2018, 76(9):ftz004.

［3］赵晓娜，朝鲁门其其格，王俊瑞. 鲍曼不动杆菌外膜蛋白A生物学功能研究进展\[J\]. 临床医药文献电子杂志，2018，5(A2):233-234.

［4］李兴国，温汉春. 鲍曼不动杆菌外膜蛋白的研究进展\[J\]. 山东医药，2017，57(29):110-112.

［5］Choi CH, Hyun SH, Lee JY, et al. Acinetobacter baumannii outer membrane protein A targets the nucleus and induces cytotoxicity\[J\]. Cell Microbiol, 2008, 10(2):309-319.

［6］Lee JS, Choi CH, Kim JW, et al. Acinetobacter baumannii outer membrane protein A induces dendritic cell death through mitochondrial targeting \[J\] . J Microbiol, 2010, 48(3): 387-392.

［7］安志远，黄骁舾，丁文一. 鲍曼不动杆菌外膜蛋白A真核表达载体构建及其诱导Hela细胞不完全自噬\[J\]. 免疫学杂志，2017，33(11):996-1002.

［8］安志远，丁文一. 鲍曼不动杆菌外膜蛋白A慢病毒载体的构建及其对RAW264.7细胞NLRP3炎症小体的激活\[J\]. 中国病原生物学杂志，2019，14(2):146-150.

［9］郑义盈，李宝宝，黄海峰，等. 多杀性巴氏杆菌OmpA基因的原核表达及生物信息学分析\[J\]. 中国畜牧兽医，2019，46(7):1926-1934.

［10］D′Arcy MS. Cell death: a review of the major forms of apoptosis， necrosis and autophagy\[J\]. Cell Biol Int, 2019, 43(6):582-592.

［11］Jin JS, Kwon SO, Moon DC, et al.Acinetobacter baumannii secretes cytotoxic outer membrane protein A via outer membrane vesicles\[J\]. PLoS One, 2011, 6(2):e17027.

［12］马星宇，狄乾乾，陈玮琳. 含pyrin结构域NOD样受体家族3(NLRP3)炎性体抑制剂及其作用机制研究进展\[J\]. 细胞与分子免疫学杂志， 2019， 35(4):373-377.

［13］Kelley N, Jeltema D, Duan Y, et al. The NLRP3 inflammasome: an overview of mechanisms of activation and regulation\[J\]. Int J Mol Sci, 2019, 20:13.

［14］吴亮，陆娟，王廷廷，等. 鲍曼不动杆菌荚膜厚度对鼠巨噬细胞炎症复合体活化的影响\[J\]. 江苏大学学报(医学版), 2019, 29(2):133-137.

［15］Liu M, Sun T, Li N, et al. BRG1 attenuates colonic inflammation and tumorigenesis through autophagydependent oxidative stress sequestration\[J\]. Nat Commun, 2019, 10(1): 4614.

［16］Deretic V, Saitoh T, Akira S. Autophagy in infection, inflammation and immunity\[J\]. Nat Rev Immunol, 2013, 13(10): 722-737.