miR-23a-3p在脂多糖诱导的人肾小管上皮细胞中的表达及作用  

Effects and role of miR-23a-3p in human renal tubular epithelial cells induced by lipopolysaccharide

作　　者：许新杰 葛勤敏[1] XU Xinjie;GE Qinmin(Department of Emergency,Xinhua Hospital Affiliated Shanghai Jiao Tong University School of Medicine,Shanghai 200092,China)

机构地区：[1]上海交通大学医学院附属新华医院急诊科,上海200092

出　　处：《同济大学学报（医学版）》2022年第6期763-768,共6页Journal of Tongji University(Medical Science)

基　　金：国家临床重点专科建设项目(2013-2014)。

摘　　要：目的 探讨miR-23a-3p在脂多糖诱导的人肾小管上皮细胞中的表达及其作用。方法 使用LPS诱导人肾小管上皮细胞HK-2细胞建立脓毒症性急性肾损伤细胞疾病模型,用CCK-8法检测细胞活力,实时定量聚合酶链反应(qRT-PCR)检测IL-6 mRNA和miR-23a-3p的相对表达量。将实验细胞分为3组:正常对照组、LPS+空白对照(NC)组和LPS+miR-23a-3p模拟物(mimic)组,采用Western印迹法检测炎症和凋亡相关蛋白p-NF-κB、IL-6、Cleaved Caspase-3和Usp5的表达水平。结果 LPS可以抑制HK-2的细胞活力并增加炎症因子IL-6 mRNA的表达水平,同时,miR-23a-3p在LPS诱导的HK-2细胞中表达下降。与正常对照组相比,LPS增加了p-NF-κB、IL-6、Cleaved Caspase-3和Usp5蛋白的表达水平,而转染miR-23a-3p模拟物可以逆转以上这些变化。结论 miR-23a-3p模拟物可以显著减轻LPS诱导的HK-2细胞的炎症和凋亡水平,其作用机制可能与靶向抑制Usp5有关。Objective To explore the effect of miR-23a-3p on induction of human renal tubular epithelial cells by lipopolysaccharide(LPS). Methods Human renal tubular epithelial HK-2 cells were induced by LPS to establish a sepsis-associated acute kidney injury model. CCK-8 was used to detect cell viability, and quantitative real-time RT-PCR(qRT-PCR) was used to detect the relative expression level of IL-6 mRNA and miR-23a-3p. HK-2 cells were divided into three groups: normal control group, LPS+blank control(NC) group, and LPS+miR-23a-3p mimic group. The expression levels of inflammation and apoptosis-related proteins were detected by Western blotting. Results LPS inhibited the viability of HK-2 cells and increased the expression level of IL-6 mRNA. Meanwhile, the expression of miR-23a-3p decreased in LPS-induced HK-2 cells. Compared with the normal control group, LPS increased the expression levels of p-NF-κB, IL-6, Cleaved Caspase-3, and Usp5 proteins;while transfection with the miR-23a-3p mimic was able to reverse these changes. Conclusion miR-23a-3p can significantly reduce LPS-induced inflammation and apoptosis in HK-2 cells, which may be related to the targeted inhibition of Usp5.

关 键 词：miR-23a-3p 脂多糖 人肾小管上皮细胞 

分 类 号：R515[医药卫生—内科学]