miR-23b表达变化对脂多糖诱导巨噬细胞炎症反应的影响

Effect of miR-23b expression on lipopolysaccharide-induced macrophage inflammatory response

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。

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 10μg/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 IKKαprotein were detected by qPCR,and the appropriate concentration of LPS was 1μg/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-1βand 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.