汉防己甲素对人脐静脉内皮细胞VEGF mRNA和Flk1蛋白的影响.docVIP

汉防己甲素对人脐静脉内皮细胞VEGF mRNA和Flk1蛋白的影响 【摘要】 目的 观察汉防己甲素对体外培养的人脐静脉血管内皮细胞(Human umbilical vEin endothelial cells , HUVEC)中血管内皮生长因子(Vascular endothelial growth factor , VEGF)mRNA 和血管内皮生长因子受体-2(Fetal liver kinase， Flk1)蛋白表达的影响,探讨其可能的作用机制。方法 应用MTT法测定不同浓度(10-3、10-4、10-5、10-6M)Tet对HUVEC的抑制作用，用免疫组织化学和原位杂交方法分别检测血管内皮生长因子受体-2(Flk1)蛋白和血管内皮生长因子(VEGF)mRNA在血管内皮细胞中的表达。结果 (1)Tet能抑制HUVEC增殖且具有量效依赖关系;(2)Flk1蛋白、VEGF mRNA在1640对照组、Tet实验组中的表达率分别为(0.742±0.057， 0.445±0.066)，(0.596±0.047， 0.324±0.049)。Tet明显降低Flk1蛋白、VEGF mRNA在内皮细胞中的表达(P0.05)。结论 汉防己甲素下调人脐静脉内皮细胞中VEGF mRNA和Flk1蛋白的表达，从而抑制人脐静脉内皮细胞增殖。 【关键词】 汉防己甲素;人脐静脉内皮细胞;血管内皮生长因子受体;血管内皮生长因子 　Flk1 protEIn and VEGF mRNA in tetrandrine on human umbilical vein endothelial cells 　　LI Dai,LI Qing-chun.Department of Ophthalmology,Xianning College, Xianning 437100,China 　　[Abstract] Objective To investigate the inhibitory effect of tetrandrine (Tet) on the VEGF mRNA and Flk1 in the third passage human umbilical vein endothelial cell (HUVEC) in vitro and discuss the drug effects of Tetrandrine in anti-angiogenesis.Methods Cultured HUVEC were exposed to different concentrations of Tet solution(10-3、10-4、10-5、10-6M) and investigated by MTT assay. Using immunochemistry to observe the expression of Flk1 and VEGF mRNA was examined by utlizing in situ hybridization (ISH) in cultured HUVEC.Results (1)Tet (10-3、10-4、10-5、10-6M) significantly suppressed the proliferation of HUVEC in vitro and showed a dose-dpendent tendency. (2)The rate of the expression of Flk1 and VEGF mRNA in Tet-treated group was 0.445±0.066， 0.324±0.049, respectively and that in control group was 0.742±0.057, 0.596±0.047，respectively.Conclusion Tetrandrine at some concentrations can suppress the expression of Flk1 and VEGF mRNA in cultured HUVEC, and inhibit HUVEC’s proliferative activity. Tetrandrine is a potent antiangiogenic agent. 　　[Key words] Tetrandrine; human umbilical vein endothelial cell; Flk1; VEGF 　　眼部新生血管是许多种眼科疾病的常见并发症之一,常导致视力减退或盲，而血管内皮细胞生长因子(VEGF)在血管生成过程中起着重要作用[1],因此近年来,新生血管研究的体