淋巴瘤细胞系Raji细胞血管内皮细胞生长因子表达及基质胶中ECV304细胞血管形成与雷公藤内酯醇的干预  

Effects of triptolide on the expression of vascular endothelial growth factor in Raji cells and angiogenesis of ECV304 cells in matrigel

作　　者：崔国惠[1] 陈卫华[1] 张纯[1] 陈燕[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院血液病研究所,湖北省武汉市430022

出　　处：《中国组织工程研究与临床康复》2008年第11期2039-2042,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30472267)~~

摘　　要：观察雷公藤内酯醇对淋巴瘤细胞系Raji细胞血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)的表达、合成分泌以及对基质胶中ECV304细胞血管形成的影响。方法:实验于2005-08/2006-02在华中科技大学同济医学院公共卫生学院分子生物学实验室完成。①MTT法检测3.125,6.25,12.5,25,50nmol/L的雷公藤内酯醇在作用12,24,36,48,60,72h对Raji细胞增殖的影响。②RT-PCR法检测12.5,25,50nmol/L雷公藤内酯醇处理Raji细胞24h后,细胞VEGFmRNA表达的变化。③ELISA法检测25nmol/L雷公藤内酯醇处理Raji细胞24h后,细胞培养上清VEGF的分泌量。④采用血管形成实验检测雷公藤内酯醇对ECV304细胞在基质胶中血管形成的影响。结果:①雷公藤内酯醇以时间和剂量依赖方式抑制Raji细胞增殖,24h半数抑制浓度为25nmol/L。②Raji细胞内VEGFmRNA主要为VEGF165和VEGF121,雷公藤内酯醇明显抑制其表达,呈剂量依赖方式。③雷公藤内酯醇明显抑制Raji细胞VEGF的分泌量(P<0.01)。④Raji细胞培养上清、10μg/L VEGF处理的Raji细胞培养上清可促进ECV304细胞在基质胶中形成血管,而25nmol/L雷公藤内酯醇处理的Raji细胞培养上清加入基质胶则未见血管形成。结论:雷公藤内酯醇以时间和剂量依赖性方式抑制淋巴瘤细胞增殖、VEGF的表达分泌及血管形成。AIM： To investigate the effect of triptolide on the expression of vascular endothelial growth factor （VEGF） in Raji cell line and on angiogenesis in cell line （ECV31M）. METHODS： The experiment was performed at the Molecular Biology Laboratory of Public Health Constitute, Tongji Medical College, Huazhong University of Science and Technology from August 2005 to February 2006. ①The antiproliferative effects of triptolide （3.125, 6.25, 12.5, 25 and 50 nmol/L） against Raji cells were determined by the MTT method at 12, 24, 36, 48, 60 and 72 hours. ②After 24 hours incubations treated by different concentrations of triptolide （12.5, 25, 50 nmol/L）, the mRNA level of VEGF by Raji cells was measured by reverse transcription-polymerase chain reaction （RT-PCR）. ③After 24 hours treatment with 25 nmol/L triptolide, VEGF concentrations in cell culture supernatants were determined by enzyme-labeled immunosorbent assay （ELISA）. （g） Effect of triptolide on angiopoiesis of ECV304 cells in matrigel was detected by angiopoiesis test. RESULTS： ①The triptolide significantly inhibited the proliferation of Raji cells in time- and dose-dependent manner. IC50 value was 25 mmol/L. ②The mRNA expression of VEGF165 and VEGF121 （containing 165 and 121 amino acid residues, respectively） was detected in all groups. Triptolide reduced the mRNA expression of VEGF165 and VEGF121 in a dose-dependent manner. ③ Triptolide significantly suppressed VEGF expression in Raji cells （P 〈 0.01）. ④There was blood vessel formation on matrigel in the supernatants from Raji culture group and VEGF group. No network and cord formed were seen in control and triptolide groups. CONCLUSION： Triptolide could inhibit the proliferation of Raji cells in time- and dose-dependent manners as well as the expressions of VEGF in Raji cells and angiogenesis in ECV304 cells.

关 键 词：雷公藤内酯醇 血管内皮细胞生长因子 血管形成 

分 类 号：R329.114[医药卫生—人体解剖和组织胚胎学]