摘要： 目的 探讨miR-181a-5p在缺氧诱导的大鼠肺动脉高压（PAH）中的作用与机制。方法 使用野百合碱（MCT）诱导建立PAH大鼠模型，使用miR-181a-5p过表达慢病毒（LV-miR-181a-5p）进行干预，利用HE染色法观察肺动脉组织的病理形态学变化。原代分离与提取大鼠肺动脉内皮细胞，通过缺氧诱导建立PAH细胞模型，在此基础上使用LV-miR-181a-5p进行干预，实时PCR技术检测细胞miR-181a-5p基因的表达；Western blotting检测细胞VEGF、MMP-2与MMP-9蛋白的表达，MTT法检测细胞增殖水平。结果 与对照组大鼠比较，模型组大鼠肺动脉组织中膜明显增厚，肺血管生成数量显著增多，内皮细胞损伤；过表达miR-181a-5p后，LV-miR181a-5p组大鼠肺动脉组织中膜增厚、肺血管生成数量增多、内皮细胞损伤现象显著改善。与对照组细胞比较，模型组细胞miR-181a-5p的表达水平显著下降（P<0.05），VEGF、MMP-2与MMP-9的蛋白表达水平及细胞增殖能力显著升高（P<0.05）；过表达miR-181a-5p后，细胞miR-181a-5p的表达水平显著上升（P<0.05），VEGF、MMP-2与MMP-9的蛋白表达水平及细胞增殖能力显著降低（P<0.05）。结论 miR-181a-5p能够降低肺动脉内皮细胞血管生成能力与增殖能力，改善缺氧诱导的大鼠PAH。

miR-181a-5p对缺氧诱导的大鼠肺动脉高压的影响

赵海燕, 王晓非

中国医科大学附属盛京医院风湿免疫科, 沈阳 110004

收稿日期:2019-02-01出版日期:2020-02-29发布日期:2019-12-26
通讯作者:王晓非E-mail:wangxf@sj-hospital.org
作者简介:赵海燕(1984-),女,主治医师,博士.
基金资助:辽宁省自然科学基金（20180551221）；辽宁省心血管系统高血压疾病转化医学研究中心建设项目（CB10）；沈阳市风湿性疾病临床医学研究中心建设项目（S512）；辽宁省博士科研启动基金（2019-BS-288）


关键词: miR-181a-5p, 大鼠, 肺动脉高压, 血管内皮生长因子, 基质金属蛋白酶-2, 基质金属蛋白酶-9
Abstract: Objective To investigate the role and mechanism of miR-181a-5p in hypoxia-induced connective tissue disease-associated pulmonary arterial hypertension. Methods A rat model of pulmonary hypertension was established by monocrotaline induction. Lentivirus-mediated over-expression of miR-181a-5p was used for intervention. The pathological morphological changes in pulmonary arteries were observed using hematoxylin and eosin staining. Rat pulmonary artery endothelial cells in the logarithmic growth phase of growth were used. Expression of miR-181a-5p was determined by real-time PCR. Protein levels of VEGF,MMP-2,and MMP-9 were detected by Western blotting. Cell proliferation was monitored using the MTT assay. Results Compared with those in the normal control group,pulmonary arteries in the model control group were significantly thickened and significant increases were observed in the frequency of pulmonary angiogenesis and endothelial cell injury. Compared with rats in the model control group,those in the miR-181a-5p overexpression group showed significantly enhanced pulmonary artery thickening,increased pulmonary angiogenesis,and significantly decreased endothelial cell injury. Compared with the normal control group,the model group showed significantly decreased expression of miR-181a-5p(P<0.05) and significantly increased expression VEGF,MMP-2,and MMP-9 proteins and proliferation ability(P<0.05). Over-expression of miR-181a-5p was associated with significant decreases in the levels of VEGF,MMP-2,and MMP-9 and proliferation(P<0.05)compared with those of the control group. Conclusion miR-181a-5p can ameliorate hypoxic-induced connective tissue disease-associated pulmonary arterial hypertension by reducing angiogenesis and proliferation of pulmonary artery endothelial cells.
Key words: miR-181a-5p, rat, pulmonary arterial hypertension, vascular endothelial growth factors, matrix metalloproteinases-2, matrix metalloproteinases-9
PDF全文下载地址:

https://journal.cmu.edu.cn/CN/article/downloadArticleFile.do?attachType=PDF&id=2436