慢性乙醇暴露通过 TLR3/NF-κB 通路诱导 H4 细胞神经炎症和雄性 C57BL/6 小鼠焦虑样行为。
Chronic ethanol exposure induces neuroinflammation in H4 cells through TLR3 / NF-κB pathway and anxiety-like behavior in male C57BL/6 mice.
机构信息
Department of Forensic Pathology, School of Forensic Medicine, China Medical University, Shenyang, Liaoning, 110122, PR China.
Department of Forensic Pathology, School of Forensic Medicine, China Medical University, Shenyang, Liaoning, 110122, PR China; The People's Procuratorate of Liaoning Province Judicial Authentication Center, Shenyang, Liaoning, 110032, PR China; Collaborative Laboratory of Intelligentized Forensic Science (CLIFS), Shenyang, Liaoning, 110032, PR China.
出版信息
Toxicology. 2020 Dec 15;446:152625. doi: 10.1016/j.tox.2020.152625. Epub 2020 Nov 5.
Chronic alcoholism has become a major public health problem. Long-term and excessive drinking can lead to a variety of diseases. Chronic ethanol exposure can induce neuroinflammation and anxiety-like behavior, and this may be induced through the Toll-like receptor 3/nuclear factor-κB (TLR3/NF-κB) pathway. Animal experiments were performed using healthy adult male C57BL/6 N mice given 10 % (m/V) or 20 % ethanol solution as the only choice of drinkable fluid for 60, 90 or 180 d. In cell culture experiments, H4 human glioma cells were treated with 100 mM ethanol for 2 d, with the TLR3 gene silenced by RNAi and NF-κB inhibited by ammonium pyrrolidine dithiocarbamate (PDTC, 10 μM). After treatment with ethanol solution for a specific time, the anxiety-like behavior of the mice was tested using the open field test and the elevated plus maze test. Western blotting was used to detect the expression of TLR3, TLR4, NF-κB, IL-1β, IL-6, and TNF-α in the mouse hippocampus and H4 cells. The expression of IL-1β, IL-6 and TNF-α in the supernatant of cell culture medium was detected by ELISA. The open field test showed a decrease in time spent in the central area, and the elevated plus maze test showed a decrease in activity time in the open arm region. These behavioral tests indicated that ethanol caused anxiety-like behavior in mice. The expression levels of TLR3, TLR4, NF-κB, IL-1β, IL-6, and TNF-α increased after ethanol exposure in both the hippocampus of mice and H4 cells. Silencing of the TLR3 gene by RNAi or inhibition of NF-κB by PDTC attenuated the ethanol-induced increase in the expression of inflammatory factors in H4 cells. These findings indicated that chronic ethanol exposure increases the expression of TLR3 and NF-κB and produces neuroinflammation and anxiety-like behavior in male C57BL/6 mice and that ethanol-induced neuroinflammation can be caused through the TLR3/NF-κB pathway.
慢性酒精中毒已成为一个主要的公共卫生问题。长期和过量饮酒可导致多种疾病。慢性乙醇暴露可诱导神经炎症和焦虑样行为，这可能是通过 Toll 样受体 3/核因子-κB（TLR3/NF-κB）途径诱导的。动物实验使用健康成年雄性 C57BL/6N 小鼠，给予 10%（m/V）或 20%乙醇溶液作为唯一可饮用的液体，分别处理 60、90 或 180d。在细胞培养实验中，用 100mM 乙醇处理 H4 人神经胶质瘤细胞 2d，用 RNAi 沉默 TLR3 基因，用铵吡啶二硫代氨基甲酸酯（PDTC，10μM）抑制 NF-κB。用乙醇溶液处理特定时间后，用旷场试验和高架十字迷宫试验检测小鼠的焦虑样行为。Western blot 检测小鼠海马和 H4 细胞中 TLR3、TLR4、NF-κB、IL-1β、IL-6 和 TNF-α的表达。用 ELISA 检测细胞培养液上清中 IL-1β、IL-6 和 TNF-α的表达。旷场试验显示中央区停留时间减少，高架十字迷宫试验显示开放臂区域活动时间减少。这些行为学试验表明，乙醇引起小鼠焦虑样行为。乙醇暴露后，小鼠海马和 H4 细胞中 TLR3、TLR4、NF-κB、IL-1β、IL-6 和 TNF-α的表达水平升高。用 RNAi 沉默 TLR3 基因或用 PDTC 抑制 NF-κB 可减弱乙醇诱导的 H4 细胞中炎症因子表达的增加。这些结果表明，慢性乙醇暴露增加 TLR3 和 NF-κB 的表达，导致雄性 C57BL/6 小鼠产生神经炎症和焦虑样行为，乙醇诱导的神经炎症可通过 TLR3/NF-κB 途径引起。
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