Klotho inhibits the activation of NLRP3 inflammasome to alleviate lipopolysaccharide-induced inflammatory injury in A549 cells and restore mitochondrial function through SIRT1/Nrf2 signaling pathway.

IF 1.4 4区 医学 Q4 PHYSIOLOGY
Yanjun Zeng, Gang Xu, Congrui Feng, Danyan Cai, Sizhi Wu, Yuanling Liu, Yuluo Chen, Wei Ma
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Abstract

Acute lung injury is a severe clinical condition constituting a major cause of mortality in intensive care units. This study aimed to investigate the role of klotho in alleviating lipopolysaccharide (LPS)-induced acute lung injury. LPS-induced acute lung injury was used to simulate the acute lung injury caused by severe pneumonia in vitro. The viability and apoptosis of A549 cells were detected by cell counting kit-8 assay and flow cytometry. The inflammatory response, oxidative stress, and mitochondrial function in A549 cells were analyzed by commercial assay kits and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl-benzimidazolyl carbocyanine iodide (JC-1) staining. The expression of apoptosis-related proteins, Sirtuin 1 (SIRT1)/nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway-related proteins, and NOD-like receptor family pyrin domain containing 3 (NLRP3) expression in A549 cells was detected by western blot. The mtDNA synthase level in A549 cells was analyzed by reverse transcription-quantitative polymerase chain reaction. The results showed that, klotho had no cytotoxic effect on A549 cells. The viability and mitochondrial function were inhibited and apoptosis, inflammatory response, and oxidative stress were aggravated in LPS-induced A549 cells, which were all reversed by klotho. Klotho activated the SIRT1/Nrf2 signaling pathway to inhibit the LPS-induced NLRP3 inflammasome activation in A549 cells. However, EX527, a SIRT1 inhibitor, attenuated the klotho effect to suppress viability and mitochondrial function and promoted apoptosis, inflammatory response, and oxidative stress of A549 cells. In conclusion, klotho inhibited the activation of NLRP3 inflammasome to alleviate LPS-induced inflammatory injury of A549 cells and restore mitochondrial function through activating the SIRT1/Nrf2 signaling pathway.

Klotho抑制NLRP3炎症小体的激活,以减轻脂多糖诱导的A549细胞炎症损伤,并通过SIRT1/Nrf2信号通路恢复线粒体功能。
急性肺损伤是一种严重的临床情况,是重症监护室死亡的主要原因。本研究旨在探讨klotho在减轻脂多糖(LPS)诱导的急性肺损伤中的作用。采用LPS诱导的急性肺损伤体外模拟重症肺炎急性肺损伤。采用细胞计数试剂盒-8法和流式细胞仪检测A549细胞的活力和凋亡。通过商业检测试剂盒和5,5’,6,6’-四氯-1,1’,3,3’-四乙基苯并咪唑基碳菁碘化物(JC-1)染色分析A549细胞的炎症反应、氧化应激和线粒体功能。通过蛋白质印迹检测A549细胞中凋亡相关蛋白、SIRT1/核因子-红系2相关因子2(Nrf2)信号通路相关蛋白和NOD样受体家族pyrin结构域3(NLRP3)的表达。采用逆转录定量聚合酶链反应检测A549细胞线粒体DNA合成酶水平。结果表明,klotho对A549细胞无细胞毒作用。LPS诱导的A549细胞活力和线粒体功能受到抑制,细胞凋亡、炎症反应和氧化应激加重,而klotho可逆转这些现象。Klotho激活SIRT1/Nrf2信号通路以抑制LPS诱导的A549细胞中NLRP3炎症小体的激活。然而,SIRT1抑制剂EX527减弱了klotho效应,以抑制A549细胞的生存能力和线粒体功能,并促进A549细胞凋亡、炎症反应和氧化应激。总之,klotho抑制NLRP3炎症小体的激活,以减轻LPS诱导的A549细胞的炎症损伤,并通过激活SIRT1/Nrf2信号通路恢复线粒体功能。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
2.30
自引率
5.60%
发文量
36
审稿时长
6-12 weeks
期刊介绍: Chinese Journal of Physiology is a multidisciplinary open access journal. Chinese Journal of Physiology (CJP) publishes high quality original research papers in physiology and pathophysiology by authors all over the world. CJP welcomes submitted research papers in all aspects of physiology science in the molecular, cellular, tissue and systemic levels. Multidisciplinary sciences with a focus to understand the role of physiology in health and disease are also encouraged. Chinese Journal of Physiology accepts fourfold article types: Original Article, Review Article (Mini-Review included), Short Communication, and Editorial. There is no cost for readers to access the full-text contents of publications.
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