[摘要]目的：探讨重组人血管内皮抑制素(YH-16)对血管内皮细胞（EC）的作用。方法：传代培养ECV304，将不同浓度的YH-16作用于生长旺盛的EC，以MTT法筛选有效浓度。以有效浓度的YH-16作用于EC，在不同时相点光镜下观察EC形态，MTT检测细胞活性，流式细胞术检测细胞周期，实时荧光定量PCR检测VEGF的表达量。结果：YH-16作用后，光镜下可见EC发生弥散性坏死。MTT检测显示吸光度降低，流式细胞术示G2期细胞比例增多，实时荧光定量PCR检测示VEGF表达量明显降低。结论：YH-16可明显抑制EC的生长增殖，可能通过抑制EC分泌VEGF，从而抑制EC增殖、迁移及血管的形成。
[关键词]重组人血管内皮抑制素；血管内皮细胞；细胞培养
[中图分类号]R96[文献标识码]A[文章编号]1008-6455（2008）05-04
Effects of recombinanted human angiostatin on vascular endothelial cell
WU Shang-min1,ZHAO Ya-nan1,YANG Li2,WANG Lu2,GUO Shu-zhong2,LIU Yin1,LIU Dan2,LU Kai-hua2
（1. Department of Plastic Surgery,Second Afiliated Hospital,Kunming Medical College, Kunming 650101, Yunnan,China.2.The Centre of Plastic Surgery,the Fourth Military Medical University, Xi'an 710032,Shaanxi,China）
Abstract:ObjectiveTo investigate the function of recombinanted human angiostatin (YH-16) on vascular endothelial cell (EC)． Methods①Different dosage of YH-16 was used in the vascular endothelial cell, and the effective concentration (10μg/ml) of YH-16 was examined by MTT method.②The test was divided into three groups randomly as YH-16 - experiment group, PBS- blank control group and RPMI-1640 medium - positive group.③In the following 24h、48h and 72h interventions, the morphous of EC was observed by photology inverted microscope, the cytoactive was detected by method of MTT and the cell cycle was investigated by flow cytometry(FLC).The expression of VEGF was dectected by real-time fluorescent quantitate polymerase chain reaction(Real-time PCR). ResultsECs were necrosis in the effect of YH-16, and many cell debris can be seen under the light microscope. The absorptance was degraded by MTT assay(P<0.05), and FLC revealed the percentage of G2 stage was mult(P<0.05).The express of VEGF was obviously degraded(P<0.05).ConclusionYH-16 can suppress the proliferation of vascular endothelial cell actively and it can suppress the DNA synthesis. Simultaneously, it can inhibit the secreting of VEGF, and suppress the prolifetation of EC , also migration and angiogenesis．
Key words: recombinant human vascular endostatin；vascular endothelial cell；cell culture
血管畸形和血管瘤的病因目前仍不明了，治疗上尚无针对性强的措施[1]。（剩余1003字）