Objective To observe the changes of miR-21 induced by lipopolysaccharide (LPS) in the inflammatory reaction of human renal cortical proximal tubular epithelial cells (HK-2 cells), and to explore the role of miR-21 by transfecting miR-21 mimics (miR-21 mimics).Methods HK-2 cells were cultured and we induced its inflammatory response by LPS.HK-2 cells were cultured, then transfected with miR-21 mimics or NC, and and we induced its inflammatory response by LPS.The cells were collected, and real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect miR-21 and IL-6 mRNA, and Western blotting was used to detect IL-6 protein.Results (1) Compared with the control group, the mRNA and protein expression of IL-6 increased at each time point in the LPS-stimulated HK-2cells (P＜0.05), and the miR-21 expression decreased (P＜0.05). (2) Compared with the cells transfected with NC mimics, the expression of miR-21 increased in HK-2 cells after transfection with miR-21 mimics for 24 h (P＜0.05).After transfection with miR-21 mimics for 24 h and 10 ug/m L LPS stimulation for 8 h, the expression of IL-6 mRNA decreased in HK-2 cells as compared with that of HK-2 cells undergoing NC mimics transfection and LPS stimulation (P＜0.05). (3) After transfection with miR-21 mimics for 24 h and LPS treatment for 12 h, the expression of miR-21 increased in HK-2 cells as compared with that of HK-2 cells undergoing NC mimics transfection and LPS stimulation (P＜0.05);after transfection with miR-21 mimics for 24 h and LPS treatment for 24 h, IL-6 protein expression decreased in HK-2 cells as compared with that of HK-2 cells undergoing NC mimics transfection and LPS stimulation of 24 h (P＜0.05).ConclusionLPS can induce inflammatory response in HK-2 cells, IL-6 expression increases, and miR-21 expression decreases.Transfection with miR-21 mimics can attenuate LPS-induced inflammatory response in HK-2 cells.%目的 观察脂多糖(LPS)诱导人肾皮质近曲小管上皮细胞炎症反应后miR-21的变化,并通过转染miR-21模拟物(miR-21 mimics)探讨miR-21的作用.方法 培养人肾皮质近曲小管上皮HK-2细胞,采用LPS诱导其炎症反应;另培养HK-2细胞,转染miR-21 mimics或NC mimics,再用LPS诱导其炎症反应.收集细胞,采用RT-qPCR法检测miR-21、IL-6 mRNA表达,Western blotting法检测IL-6蛋白表达.结果 (1)与对照细胞比较,LPS诱导的HK-2细胞各时间点IL-6 mRNA和蛋白表达均升高,miR-21表达均降低(P均＜0.05).(2)与转染NC mimics的HK-2细胞比较,转染miR-21 mimics 24 h的HK-2细胞miR-21表达升高(P＜0.05);与转染NC mimics并经LPS诱导的HK-2细胞比较,转染miR-21 mimics 24 h并经LPS诱导8 h的HK-2细胞IL-6 mRNA表达下降(P＜0.05).(3)与转染NC mimics并经LPS诱导的HK-2细胞比较,转染miR-21 mimics 24 h并经LPS诱导12 h细胞miR-21的表达上升(P＜0.05);与转染阴性对照物并经LPS诱导24 h的HK-2细胞比较,转染miR-21 mimics 24 h并经LPS诱导24 h的HK-2细胞IL-6蛋白表达下降(P＜0.05).结论 LPS可诱导人肾皮质近曲小管上皮细胞出现炎症反应,使IL-6表达上升、miR-21表达下降;转染miR-21 mimics可减轻LPS诱导的人肾皮质近曲小管上皮细胞炎症反应.
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