加减大黄虫丸干预RF/6A细胞参与血管生成的实验研究  

Experimental Research of the Modified Dahuang Zhechong Pills in the Intervention of Angiogenesis with RF/6A Cells Involved

作　　者：罗旭昇[1] 吴星伟[1] 顾青[1] 

机构地区：[1]上海交通大学附属第一人民医院,上海200080

出　　处：《世界中西医结合杂志》2011年第9期754-759,共6页World Journal of Integrated Traditional and Western Medicine

基　　金：上海市卫生局中医药重大课题基金资助(No.2006ZD01)

摘　　要：目的探讨加减大黄虫丸干预猴脉络膜-视网膜内皮细胞(RF/6A)参与血管生成的作用。方法 MTS比色法观察加减大黄虫丸对RF/6A细胞增殖的影响;Transwell小室检测其对RF/6A细胞移行的影响;Matrigel实验检测其对RF/6A细胞管腔形成的影响;Western Blot和实时定量逆转录聚合酶链反应(RT-PCR),分别检测其对RF/6A细胞表达VEGF、基质金属蛋白酶2(MMP-2)蛋白和mRNA的影响。结果 0.2 g.mL-1浓度的加减大黄虫丸对VEGF诱导的RF/6A细胞增殖具有抑制作用;其浓度为01、2.52、5和50 mg.mL-1时RF/6A细胞移行数分别为73.333±4.51、61.333±4.042、8.667±6.66和17.667±4.16;其浓度为01、2.52、5和50 mg.mL-1时RF/6A细胞内皮管腔形成数分别为20.333±0.581、3.333±1.53、11.000±1.00和1.333±0.58;其100和200 mg.mL-1浓度能够抑制RF/6A细胞VEGF、MMP-2蛋白和mRNA表达,400 mg.mL-1浓度能够抑制RF/6A细胞MMP-2蛋白和VEGF mRNA表达。结论加减大黄虫丸通过抑制RF/6A细胞增殖、移行及管腔形成的途径抑制其参与新生血管生成,其机理可能与抑制RF/6A细胞VEGF、MMP-2的表达有关。Objective To explore the effect of modified dahuang zhechong pills on the intervention of angiogenesis with monkey choroid-retinal endothelial cells（RF/6A）involved.Methods MTS assay was used to observe the impact of modified dahuang zhechong pills on the proliferation of RF/6A cells.Transwell chamber test was applied to detect its influence on the migration of RF/6A cells.Matrigel experiment was provided to detect its impacts on lumen formation of RF/6A cells.Western Blot and real-time quantitative reverse transcription-polymerase chain reaction（RT-PCR）were used to detect the expressions of VEGF,matrix metalloproteinase-2（MMP-2）and mRNA of RF/6A cells respectively.Results The modified dahuang zhechong pills,0.2 g·mL-1 inhibited the proliferation of RF/6A cells induced by VEGF.Separately,with the concentrations of 0,12.5,25 and 50 mg·mL-1,the numbers of RF/6A for cell migration were 73.333±4.51,61.333±4.04,28.667±6.66 and 17.667±4.16.With the concentration of 0,12.5,25 and 50 mg·mL-1,the numbers of RF/6A cells for lumen formation were 20.333±0.58,13.333±1.53,11.000±1.00 and 1.333±0.58.With the concentration of 100 and 200 mg·mL-1,the expressions of VEGF,MMP-2 and mRNA of RF/6A cells were inhibited.With the concentration of 400 mg·mL-1,the expressions of MMP-2 and mRNA of RF/6A cells were inhibited.Conclusion The modified dahuang zhechong pills inhibit RF/6A cells in neovascularization via the inhibition of RF/6A cell proliferation,migration and lumen formation,which is probably relevant with the inhibition of the expressions of VEGF and MMP-2 of RF/6A cells.

关 键 词：加减大黄虫丸 猴脉络膜-视网膜内皮细胞(RF/6A) 增殖 移行 脉络膜新生血管(CNV) 血管内皮生长因子(VEGF) 基质金属蛋白酶2(MMP-2) 

分 类 号：R285.5[医药卫生—中药学]