RORα在脂多糖所致小鼠巨噬细胞炎症反应中的表达变化与作用实验研究

Expression and role of RORαin lipopolysaccharide induced macrophage inflammatory response in mice

目的:探讨RORα在脂多糖所致小鼠巨噬细胞炎症反应过程中的表达变化与作用。方法:取小鼠骨髓来源的巨噬细胞(BMDMs),加入100 ng/ml脂多糖(LPS)处理不同时间点后,利用荧光定量PCR(qRT-PCR)法检测细胞中炎症因子白介素1β(IL-1β)和肿瘤坏死因子-α(TNF-α),以及RORα的转录水平变化,进一步利用Western blot检测RORα蛋白水平变化;分别采用RORα激动剂SR1078和拮抗剂SR3335预处理细胞24 h后,LPS处理2 h,利用ELISA法检测细胞上清中炎症因子IL-1β,TNF-α和IL-6的含量。结果:100 ng/ml LPS在不同时间点均可显著促进BMDMs炎症因子IL-1β和TNF-α的转录水平表达(P<0.05),而在LPS处理1~12 h显著抑制了RORα的转录水平(P<0.05),同时蛋白表达水平也明显降低(P<0.05)。与LPS处理组相比,SR1078预处理,可显著抑制炎症因子的释放,而SR3335进一步激活了细胞炎症因子的释放(P<0.05)。结论:RORα对脂多糖所致小鼠巨噬细胞炎症反应具有调节作用。

Objective:To investigate the expression and role of RORαin lipopolysaccharide induced macrophage inflammation in mice.Methods:Mouse bone marrow-derived macrophages(BMDMs)were used and treated with 100 ng/ml lipolypolysaccharide(LPS)at different time points.The transcription levels of inflammatory factors IL-1βand TNF-α,and RORαin the BMDMs were analyzed by quantitative real time PCR(qRT-PCR),and the protein level of RORαwas further detected by Western blot.The cells were pretreated with RORαagonist SR1078 and antagonist SR3335 for 24 hours respectively,and then treated with LPS for 2 hours.The concentration of IL-1β,TNF-αand IL-6 in the cell supernatant were measured by ELISA.Results:The transcription levels of IL-1βand TNF-αwere significantly upregulated in BMDMs exposed to 100 ng/ml LPS at different time points(P<0.05),while the transcription levels of RORαwere significantly inhibited after LPS treatment about 1-12 hours(P<0.05),and the protein expression levels were also significantly decreased(P<0.05).Compared with the LPS treatment group,SR1078 pretreatment significantly inhibited the release of inflammatory cytokines,while SR3335 further activated the release of inflammatory cytokines(P<0.05).Conclusion:RORαcan regulate the inflammatory response of mouse BMDMs in response to LPS.