血小板微粒通过激活单核细胞HIF-1α促进血管生成的作用  

Platelet microparticles provoke angiogenesis via activation of HIF-1α in monocytes

作　　者：贝俊杰 雷灵亮 赵芬 孙诚 农耀明 洪绍彩 赵丽霞 BEI Jun-jie;LEI Ling-liang;ZHAO Fen;SUN Cheng;NONG Yao-ming;HONG Shao-cai;ZHAO Li-xia(Department of Cardiology,Guangxi Provincial Corps Hospital of CAPF,Nanning 530003,China;Department of Car- diology,Guangzhou First People’s Hospital,Guangzhou 510180,China)

机构地区：[1]武警广西总队医院心内科,广西南宁530003 [2]广州市第一人民医院心内科,广东广州510180

出　　处：《中国病理生理杂志》2019年第10期1769-1775,共7页Chinese Journal of Pathophysiology

基　　金：广西自然科学基金资助项目(No.2015GXNSFAA139205)

摘　　要：目的:探讨血小板微粒(PMP)通过单核细胞促进血管生成的作用及机制。方法:制备人PMP,采用基质胶栓实验检测PMP对体内新生血管形成的影响。在体外常氧和缺氧条件下,使PMP与人单核细胞系THP-1结合,ELISA法检测血管内皮生长因子(VEGF)的分泌水平,RT-qPCR检测THP-1细胞VEGF和低氧诱导因子1α(HIF-1α)mRNA的表达,转录活性实验检测THP-1细胞HIF-1α的转录活性;利用共培养系统检测PMP与THP-1细胞相互作用对人脐静脉内皮细胞(HUVECs)体外管腔形成的影响。结果:PMP诱导在体基质胶栓表面形成新生血管,HIF-1α选择性抑制剂chetomin可明显减弱该效应(P<0.01)。在常氧和缺氧条件下,PMP均呈剂量依赖性地促进THP-1细胞表达和释放VEGF,上调HIF-1αmRNA的表达并增强其转录活性;阻断PMP与THP-1细胞的相互作用可以抑制THP-1细胞HIF-1α的转录活性以及VEGF的生成。PMP激活的单核细胞可促进HUVECs在体外基质胶上形成管腔,chetomin干预导致管腔的数量明显减少。结论:PMP通过激活单核细胞的HIF-1α诱导其释放VEGF,导致新生血管的形成,这可能是PMP促进动脉粥样硬化血管生成的一种新机制。AIM:To explore the role of platelet microparticles(PMP)in angiogenesis and its mechanisms involving monocytes.METHODS:Matrigel plug assay was used to evaluate the proangiogenic activity of PMP in vivo.Under normoxia and hypoxia condition,THP-1 cells were incubated with prepared PMP in vitro.ELISA and RT-qPCR were performed to measure secreted vascular endothelial growth factor(VEGF)and the mRNA expression of VEGF and hypoxia inducible factor-1α(HIF-1α)in the THP-1 cells,respectively.Transcriptional activity of HIF-1αwas detected by TransAM method.The proangiogenic effects of PMP were detected by Matrigel tube formation assay in a monocyte/endothelial co-culture model.Moreover,the effect of P-selectin neutralizing antibody and HIF-1αinhibitor were also evaluated.RESULTS:PMP induced angiogenesis in subcutaneous implantation of Matrigel in the mice,which was suppressed by chetomin,a selective inhibitor of HIF-1α(P<0.01).Under normoxia and hypoxia condition,PMP provoked release of VEGF in THP-1 cells in a time-and dose-dependent manner(P<0.05).PMP also up-regulated the mRNA expression of VEGF and HIF-1αand enhanced transcriptional activity of HIF-1α.Blockage of the interaction between PMP and THP-1 cells or inhibition of HIF-1αactivity attenuated PMP-induced VEGF production.The interaction between PMP and THP-1 cells resulted in a dramatical HIF-1α-dependent increase in capillary-like network formation of human umbilical vein endothelial cells in vitro,and this effect was attenuated by chetomin.CONCLUSION:PMP provoke the release of VEGF in monocytes,at least in part,via HIF-1αactivation,resulting in formation of new vessels,by which PMP promote angiogenesis in atherosclerosis.

关 键 词：血小板微粒 单核细胞 低氧诱导因子1Α 血管生成 

分 类 号：R363.2[医药卫生—病理学] R329.25[医药卫生—基础医学]