X盒结合蛋白1在低浓度皮质酮对巨噬细胞免疫功能调控中的作用

目的 探讨X盒结合蛋白1(X-box binding protein 1,XBP1)在低浓度皮质酮对小鼠腹腔巨噬细胞免疫功能调控中的作用.方法 分离培养成年健康雄性C57BL/6小鼠腹腔巨噬细胞,应用XBP1-RNAi慢病毒或阴性对照慢病毒感染小鼠腹腔巨噬细胞,采用荧光显微镜观察感染效率,感染3、5 d后收集巨噬细胞提取总RNA,应用RT-PCR检测小鼠腹腔巨噬细胞XBP1 mRNA表达水平和干扰效率.XBP1-RNAi慢病毒或阴性对照慢病毒感染离体培养的小鼠腹腔巨噬细胞3 d后,再应用低浓度皮质酮(50 ng/ml)处理小鼠腹腔巨噬细胞1 h,分别应用ELISA法及激光共聚焦显微镜检测小鼠腹腔巨噬细胞培养上清中TNF-α生成及其吞噬功能变化.结果 当复感染指数(multiplicity of infection,MOI)值≥6时,XBP1-RNAi慢病毒能有效地感染小鼠腹腔巨噬细胞,感染效率＞75%.应用RT-PCR检测发现XBP1-RNAi能有效地抑制小鼠腹腔巨噬细胞XBP1基因表达.通过选择性地沉默XBP1基因表达,低浓度皮质酮(50 ng/ml)对巨噬细胞吞噬功能增强的作用受到抑制,巨噬细胞的吞噬率和吞噬指数均显著下降(P＜0.01),而且对TNF-α分泌增强的作用也受到抑制,显著低于阴性对照慢病毒组(P＜0.01).结论 采用RNA干扰选择性地沉默XBP1,能有效地抑制低浓度皮质酮对巨噬细胞吞噬和TNF-α生成增强的作用.

Role of XBP1 in immune function regulation of macrophage by low concentration of corticosterone

Objective To explore the role of X-box binding protein 1 (XBP1) in immune function regulation of macrophage by low conceatration of corticosterone. Methods Peritoneal macrophages were isolated from adult male C57BL/6 mice, and then infected with XBP1-RNAi lentivirus or negative control lentivirus. The infection efficiency was observed by fluorescence microscopy. Mice peritoneal macrophages were collected in 3 and 5 d after infection and total RNA were extracted. RT-PCR analysis was performed to detect the e...