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目的 探讨雌激素相关受体α(estrogen-related receptor alpha,ERRα)对脂多糖(lipopolysaccharide,LPS)诱导大鼠肺微血管内皮细胞(pulmonary microvascular endothelial cells,PMVECs)炎症反应的影响及其机制.方法 体外培养PMVECs细胞株,当细胞处于对数生长期时利用慢病毒转染细胞,并构建稳定低表达的ERRα细胞株.将细胞分别为四组:正常对照组(Ctr组)、正常细胞+LPS处理组(Ctr+LPS组)、shERRα 1基因敲低组(shERRα 1组)和shERRα 1基因敲低组+LPS处理组(shERRα1+LPS组).予20μg/mL的LPS分别刺激对照组和基因敲低组细胞6、12和24 h后,应用cell counting kit-8(cck-8)检测各组细胞的增殖能力;应用酶联免疫吸附试验(ELISA)检测细胞培养液中的肿瘤坏死因子α(TNF-α)、白介素1β (IL-1 β)的浓度,于LPS刺激12 h后采用Western blot检测各组细胞ERRα及NF-κ B通路的相关蛋白(p-p65、p65、P-IKB α、IKBα)的表达水平.两组变量比较采用SNK-q检验,多组变量比较采用单因素方差分析,方差不齐时则用秩转换的非参数检验.以P＜0.05为差异有统计学意义.结果 与对照组相比,shERRα1组ERRα蛋白的表达量明显降低(0.09±0.01 vs 0.15±0.01);于LPS刺激6、12和24 h后,与对照组相比,shERRα1+LPS组细胞增殖能力明显降低[(99.68±4.53)％vs(48.62±1.60)％];细胞培养上清液中的TNF-α(ng/mL)、IL-1 β(ng/mL)的浓度明显升高,于刺激12h后变化最为明显(15.76±3.38 vs5 498.91±367.95;14.41±3.86 vs 6 014.92±277.33).同时,shERRα1+LPS组p-p65(0.30±0.5 vs 1.05±0.07)、p-IKBα(0.27±0.04 vs 0.77±0.06)表达量明显升高,而IKB α的表达量明显降低(0.96±0.07 vs 0.14±0.04),差异均具有统计学意义(均P＜0.05).结论 ERRα基因通过抑制NF-κ B信号通路激活,缓解LPS诱导的大鼠肺微血管内皮细胞炎症反应.“,”Objective To investigate the effect of estrogen-related receptor alpha(ERRα)on lipopolysaccharide-induced inflammatory response in rat pulmonary microvascular endothelial cells (PMVECs) and its mechanism.Methods PMVECs were cultured in vitro.When the cells were in the logarithmic growth phase,the cell were ransfected with lentivirus,and a stable low-expression ERRα cell line was constructed.The cells were divided into four groups:Ctr group (normal control group),Ctr+LPS group (normal celI+LPS treatment group),shERRα1 (shERRα1 gene knockdown group),and shERRα1+LPS group (shERRα1 gene knockdown +LPS treatment group).After 20 μg/mL LPS stimulated cells in the control group and shERRal group for 6,12 and 24 h,cell counting kit-8 (cck-8) was used to detect the cell proliferation ability of each group,and enzyme-linked immunosorbent assay (ELISA) was used to detect the concentrations of tumor necrosis factor alpha (TNF-α) and Interleukin-1β (IL-1β) in cell culture fluid.After 12 h LPS stimulation,the expression levels of ERRα and NF-κB related proteins (p-p65,p65,P-IKBα,IKBα) were measured by Western blot.Pairwise comparisons were performed with SNK-q test (two-tailed),and multiple-group comparisons were performed with one-way ANOVA.The non-parametric test of rank transformation was used when homogeneity of variance were not met.P value＜0.05 was considered significantly different.Results Compared with the control group,ERRα expression in the shERRα group was significantly decreased (0.09±0.01 vs 0.15±0.01).At 6,12 and 24 h after LPS stimulation,compared with the control group,the cell proliferation ability (％) of the shERRαl+LPS group was significantly reduced (99.68±4.53 vs 48.62±1.60) and the concentration of TNF-α (ng/mL) (15.76±3.38 vs 5 498.91±367.95) and IL-1β (ng/mL) (14.41±3.86 vs 6 014.92±277.33) in the cell culture supematant were significantly increased.The change was most obvious after 12 h stimulation.Meanwhile the expression of p-p65 (0.30±0.50 vs 1.05±0.07) and p-IKBα (0.27±0.04 vs 0.77±0.06) were increased significantly,while the expression of IKBα (0.96±0.07 vs 0.14±0.04) was decreased significantly in the shERRαl+LPS group (all P＜0.05).Conclusion ERRα gene attenuates LPS-induced inflammatory response in rat pulmonary microvascular endothelial cells by inhibiting NF-κB signaling pathway activation.