Journal Article

Brucella Melitensis 16M Regulates the Effect of AIR Domain on Inflammatory Factors, Autophagy, and Apoptosis in Mouse Macrophage through the ROS Signaling Pathway

Zhang, Hui, Wang, Zhen, Guo, Fei, Tong, Zhixia, Li, Tiansen, Huang, Meiling, Xu, Yafang, Liu, Laizhen, Chen, Chuangfu

2016

Overview

Brucellosis is a highly contagious zoonosis caused by Brucella. Brucella can invade and persist inside host cells, which results in chronic infection. We constructed AIR interference and overexpression lentiviruses to acquire AIR interference, overexpression, and rescue stable expression cell lines. We also established a Brucella melitensis 16M-infected macrophage model, which was treated with either the vehicle control or NAC (ROS scavenger N-acetylcysteine (NAC) for 0, 3, 6, 12, and 24 h. Confocal laser microscopy, transmission electron microscopy, fluorescence quantitative PCR, flow cytometry, ELISA, and Western blot were used to detect inflammation, cell autophagy and apoptosis-related protein expression levels, ROS levels, and the distribution of mitochondria. It was found that after interference and overexpression of AIR, ROS release was significantly changed, and mitochondria became abnormally aggregated. B. melitensis 16M activated the NLRP3/AIM2 inflammatory complex, and induced RAW264.7 cells to secrete IL-1β and IL-18 through the ROS pathway. B. melitensis 16M also altered autophagy-related gene expression, increased autophagy activity, and induced cell apoptosis through the ROS pathway. The results showed that after B. melitensis 16M infection, ROS induced apoptosis, inflammation, and autophagy while AIR inhibited autophagosome maturation and autophagy initiation. Autophagy negatively regulated the activation of inflammasomes and prevented inflammation from occurring. In addition, mitophagy could promote cell apoptosis.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Acetylcysteine / Analysis / Animal sciences / Animals / Apoptosis / Apoptosis - genetics / Apoptosis Regulatory Proteins - genetics / Apoptosis Regulatory Proteins - metabolism / Autophagy / Autophagy - genetics / Biology and Life Sciences / Biomarkers / Brucella / Brucella melitensis / Brucella melitensis - physiology / Brucellosis / Brucellosis - genetics / Brucellosis - metabolism / Brucellosis - microbiology / Cell death / Cell division / Cell Line / Cell lines / Cells, Cultured / Chronic infection / Cytokines - genetics / Cytokines - metabolism / Cytometry / Disease control / Electron microscopy / Enzyme-Linked Immunosorbent Assay / Flow cytometry / Fluorescence / Gene Expression / Gene Expression Profiling / Genetic aspects / Health aspects / Humans / Immune system / Infections / Inflammasomes / Inflammasomes - metabolism / Inflammation / Inflammation Mediators - chemistry / Inflammation Mediators - metabolism / Interference / Interleukin 18 / Kinases / Laser microscopy / Macrophages / Macrophages - immunology / Macrophages - metabolism / Medicine and Health Sciences / Mice / Mitochondria / Mitochondria - metabolism / Phagocytosis / Physiological aspects / Plasmids / Protein Binding / Protein Domains / Protein Transport / Proteins / Reactive oxygen species / Reactive Oxygen Species - metabolism / Research and analysis methods / Signal Transduction / Signaling / Studies / Transmission electron microscopy / Zoology