Sesn2通过激活mTOR信号通路调节LPS诱导的巨噬细胞M1极化和炎症反应  

Sesn2 modulates LPS-induced macrophage M1 polarization and inflammatory response by activating mTOR signaling pathway

作　　者：唐坤 吴黎虹 曾陈芳 党红星[1] 许峰[1] TANG Kun;WU Lihong;ZENG Chenfang;DANG Hongxing;XU Feng(Department of Pediatric Intensive Care Unit,Children's Hospital of Chongqing Medical University,National Clinical Research Center for Child Health and Disorders,Ministry of Education Key Laboratory of Child Development and Disorders,Chongqing Key Laboratory of Pediatrics,Chongqing 400014,China)

机构地区：[1]重庆医科大学附属儿童医院重症医学科,国家儿童健康与疾病临床医学研究中心,儿童发育疾病研究教育部重点实验室,儿科学重庆市重点实验室,重庆400014

出　　处：《中国病理生理杂志》2024年第4期689-696,共8页Chinese Journal of Pathophysiology

基　　金：重庆市自然科学基金面上项目(No.CSTB2022NSCQMSX0983);重庆医科大学未来医学青年创新团队发展支持计划(No.2021-W0111)。

摘　　要：目的:探讨sestrin 2(Sesn2)在巨噬细胞中的功能及其通过哺乳动物雷帕霉素靶蛋白(mTOR)通路对巨噬细胞极化和炎症的影响。方法:用小干扰RNA构建Sesn2沉默(si-Sesn2)的RAW264.7小鼠巨噬细胞胞株,通过脂多糖(LPS)处理RAW264.7细胞建立体外炎症模型。RT-qPCR评估白细胞介素1β(IL-1β)、IL-6、肿瘤坏死因子α(TNF-α)、CD86和诱导型一氧化氮合酶(iNOS)的mRNA水平;ELISA检测培养液中IL-1β、IL-6和TNF-α水平;流式细胞术确定CD86标记的M1型巨噬细胞百分比;Western blot分析mTOR、p70核糖体蛋白S6激酶(p70S6K)和真核翻译启动因子4E结合蛋白1(4EBP1)的蛋白表达水平。结果:在LPS诱导的体外炎症模型中,Sesn2在RAW264.7细胞中的表达上调,IL-1β、IL-6、TNF-α以及M1型巨噬细胞标志物CD86和iNOS的表达显著增加,伴随mTOR、p70S6K和4EBP1蛋白表达水平的升高。而si-Sesn2进一步促进了上述效应,但这些变化可通过雷帕霉素预处理减轻。结论:Sesn2可通过mTOR/p70S6K/4EBP1信号通路调控巨噬细胞M1型极化,并影响促炎因子的表达。AIM:To investigate the function of sestrin 2(Sesn2)in macrophages and its impact on macrophage polarization and inflammatory response.METHODS:Knockdown of Sesn2 in RAW264.7 mouse macrophages was established using siRNA,and an in vitro inflammation model was created by treating RAW264.7 cells with lipopolysaccharide(LPS).The mRNA levels of interleukin-1β(IL-1β),IL-6,tumor necrosis factor-α(TNF-α),CD86 and inducible nitric oxide synthase(iNOS)in RAW264.7 cells were detected by RT-qPCR.The concentrations of IL-1β,IL-6 and TNF-α in the culture medium were measured by ELISA.The percentage of M1-type macrophages marked by CD86 was determined by flow cytometry.The protein levels of mammalian target of rapamycin(mTOR),p70 ribosomal protein S6 kinase(p70S6K)and eukaryotic translation initiation factor 4E binding protein 1(4EBP1)were analyzed by Western blot.RESULTS:In the LPS-induced in vitro inflammation model,the expression levels of Sesn2,inflammatory factors(IL-1β,IL-6 and TNF-α),and M1 macrophage markers(CD86 and iNOS)in RAW264.7 cells were significantly increased,along with elevated protein expression of mTOR,p70S6K and 4EBP1.Silencing of Sesn2 with siRNA further enhanced these effects.However,if rapamycin was added before LPS in the experiment,these changes could be mitigated.CONCLUSION:Sesn2 can regulate M1 macrophage polarization through mTOR/p70S6K/4EBP1 signaling pathway and affect the expression of pro-inflammatory factors.

关 键 词：Sestrin 2 巨噬细胞极化 MTOR信号通路 炎症 

分 类 号：R364.5[医药卫生—病理学] Q257[医药卫生—基础医学] R363.2[生物学—细胞生物学]