血小板衍生膜微粒对人脐静脉内皮细胞增殖和凋亡的影响_医学论文.docVIP

血小板衍生膜微粒对人脐静脉内皮细胞增殖和凋亡的影响_医学论文 血小板衍生膜微粒对人脐静脉内皮细胞增殖和凋亡的影响_医学论文 作者：仲悦娇， 陈宝安， 黄成垠， 李翠萍， 高峰，费菲，裴孝平 【摘要】 本研究探讨血小板衍生膜微粒(platelet对人脐静脉内皮细胞（human umbilical vein endothelial cells, HUVEC）增殖和凋亡的影响。用不同浓度的凝血酶激活血小板释放出PMP，采用流式细胞术检测PMP的释放量，确定制备PMP时凝血酶的最佳应用浓度。以体外培养HUVEC为载体,通过噻唑蓝(MTT)实验和流式细胞术研究不同浓度的PMP对HUVEC增殖和凋亡的影响。结果表明，2、1.5、1.0和0.5 U/ml凝血酶激活血小板后PMP的释放率分别为28.7、47.7、50.1和43.9%；HUVEC的增殖与PMP呈剂量依赖关系。在培养液中添加40 μg/ml PMP时，HUVEC增殖是空白对照组的1.8±0.3倍；10 μl/ml 血管内皮细胞生长因子（VEGF）组的HUVEC增殖是空白对照组的1.9±0.5倍，两组之间无统计学差异，（pgt0.05）；PMP能抑制HUVEC的凋亡，40 μg/ml PMP组的细胞凋亡率为（3.9±0.4）%,明显低于空白对照组的细胞凋亡率（9.4±0.5)%（plt0.05）。VEGF（10 μl/ml）对HUVEC细胞的凋亡无明显抑制作用,其凋亡率为（8.0±0.8）%。结论： 用1 U/ml的凝血酶刺激血小板释放的PMP较为均一，且释放量最大，可促进HUVEC细胞的增殖并抑制其凋亡。 【关键词】 细胞凋亡 Effects of Platelet Proliferation and Apoptosis of Human Umbilical Vein Endothelial Cells Department of Hematology, Zhongda Hospital , Southeast University, Nanjing 210009, China 1 Jiangsu Province Blood Center, Nanjing 210042, China Abstract This study was purposed to investigate the effects of plateletderived membrane microparticles (PMP) on the proliferation and apoptosis of human umbilical vein endothelial cells (HUVEC) . Different concentrations of thrombin were adopted to activate the platelets so as to release PMPs. Flow cytometry (FCM) was adopted to evaluate the efficiencies of different concentrations of thrombin to release PMPs. By using the HUVEC cultivated in vitro as vector, the effects of PMPs on the proliferation and apoptosis of HUVEC were investigated by MTT and FCM. The results showed that the efficiencies releasing PMPs from platelets activated by 2.0, 1.5, 1.0, 0.5 U/ml thrombin were 28.7,47.7,50.1 and 43.9% respectively PMPs induced proliferation of HUVEC in a dose dependent manner. At the concentration of 40 μg/ml PMPs, the proliferation rate of HUVEC was 1.8±0.3 times as much as blank control, the proliferation rate in group of vascular endothelial growth factor was 1.9±0.5 times of as much as blank control, there was no statistic difference (pgt0.05).