MiR۲۶-۵p inhibits pathological pulmonary microvascular angiogenesis via down-regulating WNT۵A

Objective(s): Pathological micro angiogenesis is a key pathogenic factor in pulmonary diseases such as pulmonary hypertension and hepatopulmonary syndrome. More and more pieces of evidence show that excessive proliferation of pulmonary microvascular endothelial cells is the key event of pathological micro angiogenesis. The purpose of this research is to reveal the mechanism of miR۲۶-۵p regulating pulmonary microvascular hyperproliferation.Materials and Methods: Hepatopulmonary syndrome rat model was made by common bile duct ligation. HE and IHC staining were used for analysis of the pathology of the rat. CCK۸, transwell, and wound healing assay were used to assess miR۲۶-۵p or target gene WNT۵A functioned toward PMVECs. microRNA specific mimics and inhibitors were used for up/down-regulated miR۲۶-۵p expression in PMVECs. Recombinant lentivirus was used for overexpression/knockdown WNT۵A expression in PMVECs. And the regulation relationship of miR۲۶-۵p and WNT۵A was analyzed by dual-luciferase reporter assay.Results: qPCR showed that miR۲۶-۵p was significantly down-regulated in the course of HPS disease. Bioinformatics data showed that WNT۵A was one of the potential key target genes of miR۲۶-۵p. Immunohistochemistry and qPCR analysis showed that WNT۵A was largely expressed in pulmonary microvascular endothelial cells, in addition, this molecule was significantly up-regulated with the progression of the disease. Furthermore, dual luciferase reporter assay showed that miR۲۶-۵p could bind to WNT۵A ۳ ‘UTR region to inhibit WNT۵A synthesis.Conclusion: The results suggested MiR۲۶-۵p negatively regulated PMVECs proliferation and migration by WNT۵A expression. Overexpression of miR۲۶-۵p may be a potentially beneficial strategy for HPS therapy.

کلیدواژه ها:

Hepatopulmonary-syndrome (HPS) ، Migration ، miR۲۶-۵p ، Proliferation ، Pulmonary microvascular-endothelial cells (PMVECs) ، Wingless-related integration-site family member ۵A (WNT۵A)