人肺动脉平滑肌细胞中微小RNA-210通过MKP-1负性调节低氧下细胞的增殖

发布时间：2017-12-27 18:31

本文关键词：人肺动脉平滑肌细胞中微小RNA-210通过MKP-1负性调节低氧下细胞的增殖　出处：《中国病理生理杂志》2014年11期 　论文类型：期刊论文

【摘要】：目的:研究低氧条件下人肺动脉平滑肌细胞中微小RNA-210(miR-210)与丝裂原活化蛋白激酶磷酸酶1(MKP-1)的关系,是否通过MKP-1调节肺动脉平滑肌细胞增殖及其机制。方法:选用4~8代的人肺动脉平滑肌细胞,共分为12组,其中21%O2正常氧及1%O2低氧培养箱各6组,分别给予转染miR-210抑制剂、增强剂、MKP-1 siRNA等处理,提取RNA和miRNA,并采用实时定量PCR法检测各组平滑肌细胞中miR-210和MKP-1mRNA的表达,提取蛋白,并用Western blotting法比较各组MKP-1蛋白水平的表达,MTT法检测肺动脉平滑肌细胞的增殖情况。结果:低氧下,人肺动脉平滑肌细胞中miR-210及MKP-1的表达均明显增加,抑制miR-210表达使MKP-1的表达增加并可抑制低氧诱导的细胞增殖,miR-210的过表达可抑制低氧诱导的MKP-1表达上调,但不影响细胞增殖,MKP-1基因沉默后,低氧下miR-210抑制剂对细胞增殖的抑制作用消失。结论:低氧下的人肺动脉平滑肌细胞中,MKP-1是miR-210的一个新的靶基因,MKP-1可以介导miR-210抑制剂对人肺动脉平滑肌细胞增殖的负性调节作用。
[Abstract]:Objective: To study the relationship between RNA-210 (miR-210) and mitogen activated protein kinase phosphatase 1 (MKP-1) in human pulmonary artery smooth muscle cells under hypoxia, and whether MKP-1 can regulate the proliferation of pulmonary artery smooth muscle cells and its mechanism. Methods: human pulmonary artery smooth muscle cells of 4~8 generation, were divided into 12 groups, including normal 21%O2 oxygen and 1%O2 hypoxia incubator for each of the 6 groups, were treated with miR-210 inhibitor, enhanced transfection agent, MKP-1 siRNA, RNA extraction and miRNA, and were detected by real-time quantitative PCR expression, miR-210 and MKP-1mRNA were smooth muscle cell protein extraction and Western blotting method to compare the expression levels of MKP-1 protein level, the detection of proliferation of pulmonary artery smooth muscle cells by MTT. Results: under hypoxia, the expression of miR-210 and MKP-1 in pulmonary artery smooth muscle cells were significantly increased, inhibiting the expression of miR-210 increased the expression of MKP-1 and inhibits hypoxia induced cell proliferation, overexpression of miR-210 can inhibit the expression of hypoxia induced MKP-1, but did not affect cell proliferation after MKP-1 gene silencing, inhibition effect of hypoxia on the proliferation of miR-210 inhibitors. Conclusion: MKP-1 is a new target gene of miR-210 in human pulmonary artery smooth muscle cells under hypoxia. MKP-1 can mediate the negative regulation effect of miR-210 inhibitors on the proliferation of human pulmonary artery smooth muscle cells.
【作者单位】：山东大学附属省立医院儿科;
【基金】：山东省优秀中青年科学家科研奖励基金资助项目(No.BS 2011 SW 040)
【分类号】：R363
【正文快照】： 慢性低氧性肺动脉高压是临床许多心肺疾病发生发展过程中伴随或最终的病理生理环节,是一种严重的甚至危及生命的肺血管性疾病,临床上表现为肺动脉压力增高、肺血管阻力增加,右心室阻力负荷过重最终致右心衰竭,组织病理学改变为累及全层的血管炎,管壁的所有成分细胞均受累,最终

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