摘要
目的探讨IL-37对模拟失重后LPS诱导THP-1细胞炎症反应的保护作用及机制。方法细胞回转模拟失重,以细胞转染技术实现IL-37b在THP-1细胞中的过表达,以qRT-PCR、ELISA以及Western blot技术研究IL-37对模拟失重后LPS诱导THP-1细胞炎症反应的影响及机制。结果 qRT-PCR结果显示,转染IL-37b后,与未转染组相比,模拟失重后LPS刺激细胞产生的TNF-α、IL-6、IL-1β表达降低(P<0.05或P<0.01),IL-37b明显升高(P<0.01),NF-κB显著降低(P<0.05)。ELISA结果显示,转染IL-37b后,与未转染组相比,模拟失重后LPS刺激细胞释放的TNF-α、IL-6、IL-1β浓度显著下降(P<0.05)。Western blot结果表明,THP-1细胞在正常条件下可以稳定低表达IL-37,模拟失重条件下给予LPS刺激后,IL-37的表达较对照组明显升高,同时NF-κB p65的表达也明显升高;IL-37b转染THP-1细胞后,过表达的IL-37可明显抑制模拟失重条件下LPS诱导的NF-κB p65表达。结论过表达IL-37可通过下调NF-κB蛋白表达抑制模拟失重后LPS诱导的THP-1细胞炎症反应。
Objective To investigate the protective effect of IL-37 on LPS-induced inflammation in THP-1 cells under simulated weightlessness. Methods Clinostat was used to simulate microgravity in THP-1 cells with or without overexpression of IL-37 b by cell transfection technique,then qRT-PCR,ELISA and Western blot were used to explore the effects and mechanisms of IL-37 on the inflammatory response induced by lipopolysaccharide( LPS) stimulation under simulated microgravity. Results Compared with untransfected group,the inflammatory cytokines TNF-α,IL-6,IL-1β in LPS stimulated THP-1 cells after transfected with IL-37 b under simulated microgravity were decreased in both results of qRT-PCR( P〈0. 05,P〈0. 01),IL-37 b were increased significantly( P〈0. 01) and NF-κB decreased significantly( P〈0. 05). ELISA measurements showed after transfected IL-37 b,TNF-α,IL-6,IL-1β decreased significantly( P〈0. 05) in LPS stimulated THP-1 cells under simulated microgravity. Westerm blot indicated THP-1 cells might kept in stable low expression of IL-37 under normal condition. Under simulated microgravity after stimulated with LPS,the expression of IL-37 were increased significantly than that of control group and the expression of NF-κB p65 were also increased significantly. As transfected THP-1 cells with IL-37 b overexpression of IL-37 might inhibit the expression of NF-κB p65. Conclusion Overexpression of IL-37 inhibits the expression of NF-κB to alleviate the inflammatory response induced by LPS under simulated microgravity.
作者
姚静
李洁平
程江
王静宇
袁敏
郑洪伟
刘佳
袁明
Yao Jing;Li Jieping;Cheng Jiang;Wang Jingyu;Yuan Min;Zheng Hongwei;Liu Jia;Yuan Ming(The First Affiliated Hospital of Shihezi Med- ical University, Shihezi Xinjiang 832000, Chin;Yuan Ming. State Key Laboratory of Space Medicine Funda- mentals and Application, China Astronaut Research and Training Center, Beijing 100094, Chin)
出处
《航天医学与医学工程》
CAS
CSCD
北大核心
2018年第3期319-323,共5页
Space Medicine & Medical Engineering
基金
国家重点基础研究发展(973)计划(2014CB744404)
关键词
白细胞介素-37
失重模拟
脂多糖
炎症
interlukin-37
weightlessness simulation
lipopolysaccharide
inflammation