miR-33调控单核细胞系炎症因子TNF-α及IL-6表达的研究  

Regulatory Effect of miR-33 on Expression of TNF-αand IL-6 in THP-1 Cells

作　　者：赵振宇 ZHAO Zhen-yu(Department of Anesthesiology,the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine University,Changsha 410000,China)

机构地区：[1]湖南中医药大学第一附属医院麻醉科,长沙410000

出　　处：《南昌大学学报（医学版）》2020年第1期8-12,共5页Journal of Nanchang University：Medical Sciences

基　　金：国家自然科学基金青年科学基金项目(81301625)。

摘　　要：目的研究microRNA-33(miR-33)靶向人核受体相互作用蛋白1(NRIP1)负性调控脂多糖(LPS)诱导的人单核细胞(THP-1)巨噬细胞炎症反应。方法体外培养THP-1,设立:1)空白对照组(等量磷酸盐缓冲液);2)miR-33 mimics转染对照组(转染模拟miR-33 mimics);3)miR-33 mimics转染组(转染miR-33 mimics);4)miR-33 inhibitor转染对照组(转染模拟miR-33 inhibitor);5)miR-33 inhibitor转染组(转染miR-33 inhibitor)。采用10.0 ng·mL-1 LPS刺激各组细胞24 h;实时荧光定量PCR法检测细胞miR-33表达;Western印迹法检测细胞NRIP1蛋白表达;ELISA法检测细胞TNF-α、IL-6含量。结果miR-33 mimics转染组细胞NRIP1蛋白表达显著减少,TNF-α、IL-6分泌也显著降低(P<0.05);miR-33 inhibitor转染组细胞NRIP1蛋白表达显著增加,TNF-α、IL-6含量也显著增加(P<0.05)。结论miR-33可能通过下调NRIP1而抑制单核细胞TNF-α、IL-6产生负性调节炎症反应。Objective To investigate the negative regulatory effect of miRNA-33(miR-33)on lipopolysaccharide(LPS)-induced inflammatory response in THP-1 cells by targeting human nuclear receptor interaction protein 1(NRIP1).Methods The cultured THP-1 cells were divided into five groups:normal control group(treatment with same amount of phosphate buffer),miR-33 mimic transfection control group,miR-33 mimic transfection group,miR-33 inhibitor transfection control group,and miR-33 inhibitor transfection group.Cells were stimulated with 10.0 ng·mL-1 LPS for 24 hours.The expression of miR-33 was measured by real-time PCR.The expression of NRIP1 protein was detected by Western blot.The concentrations of TNF-αand IL-6 were determined by ELISA.Results The expression of NRIP1 protein and the secretion of TNF-αand IL-6 decreased in miR-33 mimic transfection group(P<0.05).However,NRIP1 expression and TNF-αand IL-6 levels increased in miR-33 inhibitor transfection group(P<0.05).Conclusion The miR-33 may negatively regulate inflammatory response by down-regulating NRIP1 expression and inhibiting TNF-αand IL-6 production in monocytes.

关 键 词：脂多糖 微小核糖核酸-33 炎症因子 人核受体相互作用蛋白1 单核细胞 

分 类 号：R392[医药卫生—免疫学]