ROS/TXNIP/NLRP3 通路介导 LPS 诱导的小胶质细胞炎症反应。
The ROS/TXNIP/NLRP3 pathway mediates LPS-induced microglial inflammatory response.
机构信息
Department of Pediatric Neurology, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325027, PR China.
Wenzhou Medical University, Wenzhou, Zhejiang 325027, PR China.
出版信息
Cytokine. 2024 Sep;181:156677. doi: 10.1016/j.cyto.2024.156677. Epub 2024 Jun 18.
BACKGROUND
Sepsis-associated encephalopathy (SAE) is a diffuse brain dysfunction activated by microglia. The potential pathological changes of SAE are complex, and the cellular pathophysiological characteristics remains unclear. This study aims to explore the ROS/TXNIP/NLRP3 pathway mediated lipopolysaccharide (LPS)-induced inflammatory response in microglia.
METHODS
BV-2 cells were pre-incubated with 10 μM N-acetyl-L-cysteine (NAC) for 2 h, which were then reacted with 1 μg/mL LPS for 24 h. Western blot assay examined the protein levels of IBA1, CD68, TXNIP, NLRP3, ASC, and Cleaved Caspase-1 in BV-2 cells. The contents of inflammatory factor were detected by ELISA assay. The co-immunoprecipitation assay examined the interaction between TXNIP and NLRP3.
RESULTS
LPS was confirmed to promote the positive expressions of IBA1 and CD68 in BV-2 cells. The further experiments indicated that LPS enhanced ROS production and NLRP3 inflammasome activation in BV-2 cells. Moreover, we also found that NAC partially reversed the facilitation of LPS on the levels of ROS, IL-1β, IL-18, TXNIP, NLRP3, ASC, and Cleaved Caspase-1 in BV-2 cells. NAC treatment also notably alleviated the interaction between TXNIP and NLRP3 in BV-2 cells.
CONCLUSION
ROS inhibition mediated NLRP3 signaling inactivation by decreasing TXNIP expression.
背景
脓毒症相关性脑病（SAE）是一种由小胶质细胞激活的弥漫性脑功能障碍。SAE 的潜在病理变化复杂，细胞病理生理特征尚不清楚。本研究旨在探讨 ROS/TXNIP/NLRP3 通路介导的脂多糖（LPS）诱导小胶质细胞炎症反应。
方法
BV-2 细胞先用 10 μM N-乙酰-L-半胱氨酸（NAC）孵育 2 h，然后用 1 μg/mL LPS 孵育 24 h。Western blot 检测 BV-2 细胞中 IBA1、CD68、TXNIP、NLRP3、ASC 和 Cleaved Caspase-1 的蛋白水平。ELISA 检测炎症因子含量。免疫共沉淀检测 TXNIP 与 NLRP3 的相互作用。
结果
LPS 被证实可促进 BV-2 细胞中 IBA1 和 CD68 的阳性表达。进一步的实验表明，LPS 增强了 BV-2 细胞中 ROS 的产生和 NLRP3 炎性小体的激活。此外，我们还发现 NAC 部分逆转了 LPS 对 BV-2 细胞中 ROS、IL-1β、IL-18、TXNIP、NLRP3、ASC 和 Cleaved Caspase-1 水平的促进作用。NAC 处理还显著减轻了 BV-2 细胞中 TXNIP 与 NLRP3 之间的相互作用。
结论
ROS 抑制通过降低 TXNIP 表达介导 NLRP3 信号失活。
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