Mo Jianwen, Zhang Daifen, Yang Renze
Department of Orthopedics Surgery, the First Affiliated Hospital of Gannan Medical University, Ganzhou 341000, Jiangxi Province, P.R. China
Department of Orthopedics Surgery, the First Affiliated Hospital of Gannan Medical University, Ganzhou 341000, Jiangxi Province, P.R. China.
Biosci Rep. 2016 Oct 14;36(5). doi: 10.1042/BSR20160139. Print 2016 Oct.
Deep vein thrombosis (DVT) is a common type of venous thrombosis. Successful resolution of DVT-related thrombi is important in the treatment of DVT. Endothelial progenitor cells (EPCs) have emerged as a promising therapeutic choice for DVT-related thrombus resolution; however, the clinical application of EPCs faces many challenges. In the present study, the expression of miR-582, miR-195 and miR-532 under hypoxic or normoxic conditions was measured using quantitative real-time PCR analysis (qRT-PCR) and the results showed that the increased fold of miR-195 was highest in human EPCs (hEPCs) under hypoxic conditions. Then the role and regulating mechanism of miR-195 in improving the function of EPCs was investigated. To investigate the effect of miR-195 inhibition on the autophagy of hEPCs, the expression of the autophagy-related genes LC3B and beclin1 was examined using western blotting, and the formation of autophagosomes was observed using TEM. The results indicated that the inhibition of miR-195 expression could promote autophagy of hEPCs. In addition, we investigated the role of miR-195 on the proliferation, migration and angiogenesis of hEPCs under hypoxia. The results revealed that miR-195 inhibition promotes cell proliferation, migration and angiogenesis of hEPCs under hypoxia. Furthermore, GABA type A receptor associated protein like 1 (GABARAPL1) was identified as a directed target of miR-195 and GABARAPL1 silencing could decrease the effect of miR-195 knockdown on cell proliferation, migration, angiogenesis and autophagy of hEPCs under hypoxia. Together, these results indicate that miR-195 regulates cell proliferation, migration, angiogenesis and autophagy of hEPCs by targeting GABARAPL1.
深静脉血栓形成（DVT）是一种常见的静脉血栓形成类型。成功溶解与DVT相关的血栓在DVT治疗中很重要。内皮祖细胞（EPCs）已成为溶解DVT相关血栓的一种有前景的治疗选择；然而，EPCs的临床应用面临许多挑战。在本研究中，使用定量实时PCR分析（qRT-PCR）测量了缺氧或常氧条件下miR-582、miR-195和miR-532的表达，结果显示在缺氧条件下，人EPCs（hEPCs）中miR-195的增加倍数最高。然后研究了miR-195在改善EPCs功能中的作用及其调控机制。为了研究miR-195抑制对hEPCs自噬的影响，使用蛋白质免疫印迹法检测自噬相关基因LC3B和beclin1的表达，并使用透射电子显微镜观察自噬体的形成。结果表明，抑制miR-195表达可促进hEPCs的自噬。此外，我们研究了miR-195在缺氧条件下对hEPCs增殖、迁移和血管生成的作用。结果显示，抑制miR-195可促进缺氧条件下hEPCs的细胞增殖、迁移和血管生成。此外，γ-氨基丁酸A型受体相关蛋白样1（GABARAPL1）被确定为miR-195的直接靶点，沉默GABARAPL1可降低miR-195敲低对缺氧条件下hEPCs细胞增殖、迁移、血管生成和自噬的影响。总之，这些结果表明miR-195通过靶向GABARAPL1调节hEPCs的细胞增殖、迁移、血管生成和自噬。
