急性肾损伤模型中miRNA-21对肾小管上皮细胞保护作用的研究

发布时间：2019-02-22 18:06

【摘要】：为了研究马兜铃酸(AA)模拟的肾小管上皮细胞系HK-2细胞急性肾损伤(AKI)过程中,miRNA-21(miR-21)和肾损伤分子-1(KIM-1)之间的调控关系,以及探讨miR-21对AA引起的AKI的影响,试验选用HK-2细胞,采用不同浓度的30,60,120μmol/L AA处理24 h后,提取各组细胞总RNA,经过逆转录后进行实时荧光定量检测KIM-1 mRNA和miR-21的转录水平。为了确认miR-21是否对KIM-1具有转录后调控作用,通过将miR-21模拟物转染进入HK-2细胞,实时荧光定量PCR检测KIM-1的mRNA转录水平,同时检测miR-21模拟物的表达,探讨miR-21对AA引起的急性肾损伤细胞是否具有保护作用。结果表明:与未处理的对照组相比,AA对HK-2细胞处理后可以促进KIM-1 mRNA和miR-21的过表达(P0.05)。而高浓度的AA(120μmol/L)则促进KIM-1 mRNA表达明显下降,而miR-21仍上调表达。miR-21模拟物的过表达,明显下调了KIM-1 mRNA的表达水平。低浓度的AA处理HK-2细胞后,miR-21和KIM-1表达均上调。当AA浓度达到一定程度后,上调的miR-21开始抑制KIM-1 mRNA的表达,最终通过负反馈调节产出作用。说明高浓度AA通过调控miR-21表达来抑制KIM-1 mRNA的表达水平,从而对AKI起到保护作用。
[Abstract]:To investigate the regulatory relationship between miRNA-21 (miR-21) and renal injury molecule-1 (KIM-1) during acute renal injury of HK-2 cells simulated by aristolochic acid (AA). To explore the effect of miR-21 on AKI induced by AA, HK-2 cells were used in the experiment. After 24 h treatment with different concentrations of 30c60120 渭 mol/L AA, the total RNA, of each group was extracted. The transcription levels of KIM-1 mRNA and miR-21 were measured by real-time fluorescence quantitative analysis after reverse transcription. In order to confirm whether miR-21 has posttranscriptional regulation on KIM-1, the mRNA transcription level of KIM-1 was detected by real-time fluorescent quantitative PCR, and the expression of miR-21 mimics was detected by transfection of miR-21 mimics into HK-2 cells. To investigate the protective effect of miR-21 on acute renal injury induced by AA. The results showed that AA could promote the overexpression of KIM-1 mRNA and miR-21 in HK-2 cells compared with untreated control group (P0.05). However, high concentration of AA (120 渭 mol/L) significantly decreased the expression of KIM-1 mRNA, while miR-21 still up-regulated the expression of miR-21 mimics, which significantly down-regulated the expression of KIM-1 mRNA. After HK-2 cells were treated with low concentration of AA, the expression of miR-21 and KIM-1 were up-regulated. When the concentration of AA reached a certain level, the up-regulated miR-21 began to inhibit the expression of KIM-1 mRNA, and finally regulated the output by negative feedback. It suggested that high concentration of AA could inhibit the expression of KIM-1 mRNA by regulating the expression of miR-21, thus it could protect AKI.
【作者单位】：西南民族大学生命科学与技术学院;四川大学华西公共卫生学院;
【分类号】：S852.3

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