沉默肝脏X受体β表达对BV2小胶质细胞极化的影响  

Effects of silencing liver X receptor β expression on BV2 microglia polarization

作　　者：吕柯毅 张丹丹 骆燚 陈胜 姜鑫 范晓棠 陈红 LYU Keyi;ZHANG Dandan;LUO Yi;CHEN Sheng;JIANG Xin;FAN Xiaotang;CHEN Hong(Key Laboratory of Cognition and Personality,Ministry of Education,Faculty of Psychology,Southwest University,Chongqing 400715,China;Department of Military Cognitive Psychology,Faculty of Medical Psychology,Army Medical University,Chongqing 400038,China)

机构地区：[1]西南大学心理学部认知与人格教育部重点实验室,重庆400715 [2]陆军军医大学医学心理系军事认知心理学教研室,重庆400038

出　　处：《免疫学杂志》2022年第2期93-99,115,共8页Immunological Journal

摘　　要：目的探讨沉默肝脏X受体β(LXRβ)对BV2小胶质细胞M1/M2表型变化和炎症因子释放的影响。方法通过RNA干扰技术沉默小鼠小胶质细胞株(BV2)中LXRβ的表达。实验分为siRNA-NC组(siRNA无关序列对照组)、siRNA-LXRβ组(转染siRNA-LXRβ质粒)。转染siRNA-LXRβ后,免疫荧光方法检测LXRβ和Iba1的表达水平;实时荧光定量PCR技术检测LXRβ、炎症因子(TNF-α、IL-6、IL-1β)、小胶质细胞M1型标记物(COX-2、iNOS、CD68)和M2型标记物(Arg-1、CD206)的mRNA水平表达。流式细胞术(flow cytometry)检测小胶质细胞M1型标记物(MHCⅡ、CD36)和M2型标记物(Arg-1、CD206)的表达水平。结果对BV2细胞分别转染24 h、48 h后发现,48 h的siRNA-LXRβ组的LXRβ蛋白及mRNA水平均显著低于NC组(P<0.001)。沉默LXRβ的表达后BV2细胞表现为胞体变圆变大,呈阿米巴样活化状态,炎症因子相关基因TNF-α、IL-6的mRNA表达水平显著升高(P<0.01),同时小胶质细胞M1型标记物(COX-2、CD68)和M2型标记物(Arg-1、CD206)的m RNA表达水平也显著升高(P<0.05);此外,流式细胞术结果显示沉默LXRβ的表达后小胶质细胞M1型标记物MHCⅡ、M2型标记物(Arg-1、CD206)在BV2小胶质细胞的比例中显著升高(P<0.01),同时M1/M2比率也明显增多(P<0.01)。结论沉默LXRβ表达后引起BV2小胶质细胞激活及炎症因子释放增多,促进BV2小胶质细胞向M1型、M2型转化,同时显著升高BV2小胶质细胞中M1/M2的比率。This study was performed to investigate the effect of silencing liver X receptorβ(LXRβ)on the alteration of M1/M2 phenotypes and the secretion of inflammatory cytokines.The expression of LXRβin mouse microglia cell line(BV2)was silenced by RNA interference.The experimental groups were divided into siRNA-NC group(siRNA of irrelevant sequence)and siRNA-LXRβgroup(transfected with siRNA-LXRβ).After transfection of siRNA-LXRβ,the expressions of LXRβand Iba1 were detected by immunofluorescence staining.Quantitative real-time PCR(qPCR)was used to detect the levels of LXRβ,inflammatory factors(TNF-α,IL-6 and IL-1β),microglia M1(COX-2,iNOS and CD86)and M2(Arg-1 and CD206)mRNA expression;flow cytometry was used to detect microglia M1(MHCⅡand CD36)and M2(Arg-1 and CD206)markers.After BV2 was transfected with siRNA LXRβfor 24 h or 48 h,the expressions of LXRβprotein and mRNA in 48 h siRNA-LXRβgroup were significantly lower than those the NC group(P<0.001).After LXRβwas silenced,BV2 cells became round and enlarged,and showed amoeba-like activation,Furthermore,the mRNA expression levels of inflammatory cytokines TNF-αand IL-6 were significantly higher(P<0.01);and the mRNA expression of M1 marker COX-2 and CD68,M2 marker Arg-1(P<0.01)and CD206 were also significantly higher(P<0.05).In addition,flowcytometry indicated that M1-marker MHCⅡ,M2-marker Arg-1 and CD206 were significantly up-regulated(P<0.01)in BV2 microglia cells after LXRβwas silenced,and the ratio of M1/M2(P<0.01)was also significantly increased.Taken together,silencing LXRβcan activate BV2 microglia and release inflammatory cytokines,promote the transformation of BV2 microglia into M1 and M2,and significantly increase the ratio of M1/M2 in BV2 microglia.

关 键 词：肝脏X受体β BV2小胶质细胞 M1 M2 

分 类 号：R392.1[医药卫生—免疫学]