摘要
目的探讨血小板衍生膜微粒(PMP)对人脐静脉内皮细胞(HUVEC)增殖和鸡胚绒毛尿囊膜(CAM)血管生成的影响。方法在血小板悬液中加1.0U/ml的凝血酶,37℃下作用10min后,800g离心20min,取上清,17000g离心分离出PMP,采用BCA法测定PMP含量(以蛋白含量计),将HUVEC与不同浓度的PMP和血管内皮细胞生长因子(VEGF)共同孵育培养24h后,采用MTT法测定细胞增殖;将20μl的PMP(40μg/ml)和VEGF(10μl/ml)分别接种于CAM上,孵育72h后,观察PMP对CAM血管生成的影响。结果HUVEC的增殖与PMPs呈剂量依赖关系,在培养液中添加40μg/mlPMP时,HUVEC增殖是空白对照组的(1.83±0.33)倍;而VEGF(10μl/ml)组HUVEC的增殖是空白对照组的(1.91±0.47)倍,两者相比差异无显著性(P>0.05)。PMP组及VEGF组在接种点周围可见不同程度放射状密集生长的血管网,而对照组无特异性密集血管生成。PMP组及VEGF组的CAM血管分支点数分别为112.5±11.3、128.4±10.0,均显著高于空白对照组82.6±8.1。结论PMP可促进人脐静脉内皮细胞的增殖和CAM血管的生成。
Objective To investigate the effect of platelet-derived microparticles（PMP） on proliferation of human umbilical vein endothelial cells （HUVEC） and angiogenesis in chick chorioallantoic membranes （CAM）. Methods The platelet concentrates were activated by 1 U/ml thrombin for 10 minutes and centrifuged at 800 g for 20 minutes,and supernatants enriched in PMP were collected by centrifugation at 17 000 g for 1 h. The protein concentration was measured using BCA method. HUVEC were incubated for 24 h with or without （control） PMP or VEGF,the effect of PMP on cell proliferation was analyzed by MTT assay. PMP were put into 9 day chick embryo. Chicken CAMs angiogenesis were observed at 72 h after incubation. Result PMP stimulated HUVEC proliferation. After 24 h of incubation,PMP stimulated HUVEC proliferation in a dose-dependent manner. At the concentration of 40 μg/ml PMP,the proliferation rate of HUVEC was as 1.83±0.33 times as control and there was no difference with the group of vascular endothelial growth factor （VEGF）, which the proliferation rate was 1.91＋0.47 times vs. control （P〉0.05）. Better effects were obtained in the formation of capillary and the count of blood vessel in groups treated by PMP,compared with the NS control group（P〈0. 05） and compared with the VEGF（P〉0.05）. Conclusion PMP could stimulate proliferation of HUVEC,and induce the formation of new blood vessels.
出处
《临床输血与检验》
CAS
2007年第3期229-232,共4页
Journal of Clinical Transfusion and Laboratory Medicine
关键词
血小板衍生膜微粒
人脐静脉内皮细胞
增殖
Platelet-derived microparticles Human umbilical vein endothelial cells Proliferation
