【摘要】 目的探讨微小RNA-23b(miR-23b)表达变化对脂多糖(LPS)诱导的巨噬细胞炎症反应的影响。方法体外培养巨噬细胞株J774A.1,分别给予0、0.1、0.5、1、5、10μg/mL的LPS处理12 h,光镜观察细胞形态变化,采用qPCR法检测NF-κB p50、TNF-α、IL-1β、IL-6 mRNA,Western blotting法检测IKKα蛋白,明确LPS适宜给药浓度为1μg/mL。将巨噬细胞分为miR-23b mimics组、阴性对照1组和miR-23b inhibitor组、阴性对照2组,分别转染miR-23b mimics、mimics control、miR-23b inhibitor、inhibitor control,之后加入1μg/mL的LPS继续培养12 h。采用qPCR法检测各组巨噬细胞中的NF-κB p50、TNF-α、IL-1β、IL-6。结果 miR-23b mimics组细胞中NF-κB p50、TNF-α、IL-1β、IL-6 mRNA相对表达量低于阴性对照1组(P均<0.01)。miR-23b inhibitor组细胞中NF-κB p50、TNF-α、IL-1β、IL-6 mRNA相对表达量高于阴性对照2组(P均<0.01)。结论 miR-23b过表达可抑制LPS诱导的巨噬细胞炎症反应,降低细胞中炎症因子表达;抑制miR-23b表达可促进LPS诱导的巨噬细胞炎症反应,增加细胞中炎症因子表达。更多还原

【Abstract】 Objective To investigate the effect of microRNA-23 b(miR-23 b) on macrophage inflammation induced by lipopolysaccharide(LPS). Methods Macrophage cell line J774 A.1 was cultured in vitro, and treated with 0, 0.1, 0.5, 1, 5, and 10 μg/mL LPS for 12 h, and the morphology of the cells was observed by light microscopy. TNF-κB p50, TNF-α, IL-1β, IL-6 mRNA and IKKα protein were detected by qPCR, and the appropriate concentration of LPS was 1 μg/mL. The macrophages were divided into the miR-23 b mimics group, negative control group 1, miR-23 b inhibitor group, and negative control group 2, respectively, and then were transfected with miR-23 b mimics, miR-23 b inhibitor, mimics control, and inhibitor control, followed by 1 ug/mL LPS culture for 12 h. Macrophage morphological changes were observed under microscope. NF-κB p50, TNF-α, IL-1β, and IL-6 were detected by qPCR. Results The relative expression of NF-κB p50, TNF-α, IL-1β and IL-6 mRNA in the miR-23 b mimics group was lower than that in the negative control group 1(all P<0.01). The relative expression of NF-κB p50, TNF-α, IL-1β, and IL-6 mRNA in the miR-23 b inhibitor group was higher than that in the negative control group 2(all P<0.01). Conclusions Overexpression of miR-23 b inhibits LPS-induced macrophage inflammatory response and reduces the expression of inflammatory factors in macrophages. Inhibition of miR-23 b can result in the increased expression of cellular inflammatory factors in LPS-induced inflammatory response.更多还原