PRDX1对脂多糖诱导的小胶质细胞炎症反应及细胞凋亡的作用机制  

PRDX1 modulates LPS-triggered inflammatory response and cell apoptosis in microglial cells through TLR4 signaling axis

作　　者：陈旭狮 姚仕奋[1] 蔡宏华[1] 罗思东 王业杨 CHEN Xushi;YAO Shifen;CAI Honghua;LUO Sidong;WANG Yeyang(The 2nd Department of Orthopedics,Huizhou Central People's Hospital,Huizhou 516008,Guangdong,China;Orthopedic Center,Guangdong Second Provincial General Hospital,Guangzhou 510000,Guangdong,China)

机构地区：[1]惠州市中心人民医院骨科二区,惠州516008 [2]广东省第二人民医院骨科中心,广州510000

出　　处：《医学研究与战创伤救治》2024年第5期456-461,共6页Journal of Medical Research & Combat Trauma Care

基　　金：广东省中医药局科研课题面上项目(20221474);惠州市科技计划项目(210418234575032)。

摘　　要：目的旨在探讨PRDX1对脂多糖诱导的小胶质细胞BV2神经炎症损伤的作用及潜在机制。方法对于PRDX1 shRNA慢病毒感染实验,BV2细胞分为:对照组(BV2细胞进行正常培养),脂多糖处理组,NCshRNA慢病毒感染组(细胞经NCshRNA慢病毒感染48 h后,用脂多糖处理24 h组),PRDX1 shRNA慢病毒感染组(细胞经PRDX1 shRNA慢病毒感染48 h后,用脂多糖处理24 h组)。对于TLR4过表达实验,在慢病毒感染实验的基础上增加TLR4过表达组(细胞经PRDX1 shR⁃NA慢病毒感染及TLR4过表达慢病毒感染48 h后,用脂多糖处理24 h)。RT-qPCR和Westernblot分别用于检测mRNA和蛋白表达水平;采用ELISA法检测细胞培养上清液中炎症因子TNF-α和IL-6的分泌水平;MTT法检测细胞增殖活性;流式细胞术检测细胞凋亡率。结果ELISA检测结果显示,与脂多糖处理的BV2细胞相比,PRDX1 shRNA慢病毒感染组TNF-α和IL-6的含量均显著减少(P<0.01)。RT-qPCR结果表明,TNF-α和IL-6的mRNA表达水平和ELISA检测结果趋势一致。MTT法检测结果表明,与对照组相比,脂多糖处理组BV2细胞活力显著降低;而与脂多糖处理组相比,PRDX1 shRNA慢病毒感染组细胞活力明显升高。流式细胞术检测结果表明,与对照组相比,脂多糖处理组BV2细胞凋亡率明显增加,而与脂多糖处理组相比,PRDX1 shRNA慢病毒感染组细胞凋亡率明显减少。Westernblot结果显示,与对照组细胞相比,脂多糖处理组BV2细胞中TLR4表达显著上调(P<0.05);而与脂多糖处理组BV2细胞相比,PRDX1 shRNA慢病毒感染显著抑制脂多糖诱导的TLR4表达(P<0.05)。与PRDX1 shRNA慢病毒感染组相比,TLR4过表达组BV2细胞中炎症因子TNF-α及IL-6水平显著升高(P<0.05),细胞活力水平降低(P<0.05),且细胞凋亡水平显著升高(P<0.05)。结论沉默PRDX1明显促进脂多糖刺激BV2细胞活力,抑制脂多糖诱导的凋亡及炎症应答反应,其可能是通过正向调控TLR4来实现的。阐明PRDX1/TLR4信号轴在脂多�Objective To investigate the roles and potential mechanism of PRDX1 on inflammatory responses and in lipopolysaccharide(LPS)-induced BV2 cells.Methods BV2 cells were divided into control group(BV2 cells were normally cultured),LPS treated group,NC shRNA lentivirus infection group(cells were infected with NC shRNA lentivirus for 48 hours and then treated with LPS for 24 hours),and PRDX1 shRNA lentivirus infection group(cells were infected with PRDX1 shRNA lentivirus for 48 hours and then treated with LPS for 24 hours).For the TLR4 overex-pression experiment,a TLR4 overexpression group was added on the basis of the lentivirus infection experiment(cells were infected with PRDX1 shRNA lentivirus and TLR4 overexpression lentivirus for 48 hours,and then treated with LPS for 24 hours).RT-qPCR and Western blot were adopted to estimate the mRNA and protein expression,respectively.The evaluation of cell viability was executed by MTT assay.Cell apoptosis was appraised by flow cytometry.ELISA assays were implemented for the measurement of the release of inflammatory factors such as TNF-αand IL-6.Results Test results of ELISA showed that compared with BV2 cells treated with LPS,the levels of TNF-αand IL-6 in the PRDX1 shRNA lentivirus infec-tion group were significantly reduced(P<0.01).The RT qPCR results showed that the mRNA expression levels of TNF-αand IL-6 were consistent with the trend of ELISA test results.The MTT assay results showed that compared with the control group,the viability of BV2 cells in the LPS treatment group was significantly reduced;Compared with the LPS treatment group,the PRDX1 shRNA lentivirus infection group showed a significant increase in cell viability.The results of flow cytometry analysis showed that compared with the control group,the apoptosis rate of BV2 cells in the LPS treatment group was significantly increased,while compared with the LPS treatment group,the apoptosis rate of PRDX1 shRNA lentivirus infection group was significantly reduced.Western blot results showed that compared with

关 键 词：PRDX1/TLR4轴 炎症应答 凋亡 脊髓损伤 

分 类 号：R364[医药卫生—病理学] R744[医药卫生—基础医学]