MiR145-5p inhibits proliferation of PMVECs via PAI-1 in experimental hepatopulmonary syndrome rat pulmonary microvascular hyperplasia
MiR145-5p inhibits proliferation of PMVECs via PAI-1 in experimental hepatopulmonary syndrome rat pulmonary microvascular hyperplasia
复制标题
MiR145-5p通过PAI-1抑制实验性肝肺综合征大鼠肺微血管增生中PMVEC的增殖
DOI：
10.1242/bio.044800
复制
发表时间：
2019-11-01
期刊：
影响因子：
2.4
通讯作者：
Yi, Bin
中科院分区：
文献类型：
--
作者：
Chen, Yang;Yang, Congwen;Yi, Bin
肝硬化综合征（HPS）是一种晚期肝病、肺内血管扩张和动脉低氧血症三联征。越来越多的证据表明HPS与肺微血管增生有关。本研究旨在探讨miR-145通过纤溶酶原激活物抑制剂-1（派-1）调控HPS肺微血管内皮细胞（PMVECs）增殖和血管生成的机制。为了测试这一点，通过苏木精和伊红（H&E）染色在来自患有HPS的大鼠的肺组织中评估肺微血管的形态评分和数目。在来自HPS大鼠的肺组织中以及在用HPS大鼠血清处理的PMVEC中评估派-1的表达水平。我们还通过生物信息学分析筛选了派-1上可能的microRNA结合位点。由于miR 145 - 5 p是派-1上的microRNA结合位点，因此通过qRT-PCR检测大鼠肺和PMVEC中miR 145 - 3 p和miR 145 - 5 p的表达水平。此外，通过上调和下调miR-145- 5 p来检测miR-145 - 5 p调控对派-1的影响，并使用特异性慢病毒转染来过表达和敲低派-1以评估派-1对PMVEC增殖的功能。我们的数据表明，派-1在大鼠肺组织中的表达水平显着增加时，大鼠胆总管结扎治疗。我们发现miR-145- 5 p的水平在HPS组织和细胞系中频繁下调，并且miR-145- 5 p的过表达显著抑制PMVEC增殖。我们进一步证实派-1是HPS中miR-145- 5 p的一个新的直接靶点。miR-145- 5 p抑制派-1的合成，派-1的表达变化直接影响PMVECs的增殖。我们得出结论，miR-145- 5 p通过派-1表达负调控PMVEC增殖。此外，miR-145- 5 p的过表达可能证明作为HPS治疗的治疗策略是有益的。总结：我们的研究结果提供了原则证据，即microRNA可能对HPS新治疗策略的未来发展有用。
ABSTRACT Hepatopulmonary syndrome (HPS) is a triad of advanced liver disease, intrapulmonary vasodilatation and arterial hypoxemia. Increasing evidence shows that HPS is associated with pulmonary microvascular hyperplasia. The aim of this work was to investigate the underlying mechanism of miR-145 in regulating the proliferation of pulmonary microvascular endothelial cells (PMVECs) and angiogenesis in HPS via plasminogen activator inhibitor-1 (PAI-1). To test this, morphology score and number of pulmonary microvascular were assessed in lung tissues from rats with HPS by Hematoxylin and Eosin (H&E) staining. Expression levels of PAI-1 were assessed in lung tissues from HPS rats, as well as in PMVECs treated with HPS rat serum. We also selected the putative microRNA binding site on PAI-1 by bioinformatics analysis. Then, miR145-3p and miR145-5p expression levels in the lungs and PMVECs of rats were detected by qRT-PCR because miR145-5p is a microRNA binding site on PAI-1. In addition, the effects of miR-145-5p regulation on PAI-1 were examined by upregulation and downregulation of miR-145-5p and specific lentivirus transfection was used to overexpress and knockdown PAI-1 to assess PAI-1 function on PMVECs proliferation. Our data showed that levels of PAI-1 expression in lung tissue of rats increased significantly when rats were treated with common bile duct ligation. We found that levels of miR-145-5p were frequently downregulated in HPS tissues and cell lines, and overexpression of miR-145-5p dramatically inhibited PMVECs proliferation. We further verified PAI-1 as a novel and direct target of miR-145-5p in HPS. MiR-145-5p inhibits PAI-1 synthesis and the expression changes of PAI-1 directly affect the proliferation of PMVECs. We concluded that miR-145-5p negatively regulates PMVEC proliferation through PAI-1 expression. In addition, overexpression of miR-145-5p may prove beneficial as a therapeutic strategy for HPS treatment. Summary: Our findings provide proof of principle that microRNAs may be useful for the future development of novel therapeutic strategies in HPS.