IL-37对模拟失重后脂多糖诱导THP-1细胞炎症反应的保护作用

Protective Effect of IL-37 on LPS Induced Inflammation in THP-1 Cells under Simulated Weightlessness

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.