复方血栓通对猴脉络膜一视网膜内皮细胞增生、移行和管腔形成的作用  

Effects of Complex Xueshuantong on the proliferation, migration, and tube formation of rhesus choroid-retina endothelial cells

作　　者：罗旭昇[2] 吴星伟[1] 顾青[1] 张红梅[1] 施宇华[1] 仇亚婷[1] 

机构地区：[1]上海交通大学附属第一人民医院眼科,上海200080 [2]中国中医科学院眼科医院眼科工作

出　　处：《中华眼视光学与视觉科学杂志》2011年第4期263-268,共6页Chinese Journal Of Optometry Ophthalmology And Visual Science

基　　金：上海市卫生局中医药重大课题资助项目（2006ZD01）

摘　　要：目的探讨复方血栓通干预猴脉络膜．视网膜内皮细胞（Rr／6A）参与血管生成的作用及机理。方法实验研究。本研究分为四部分：①取对数生长期RF／6A细胞进行培养，设立空白对照组、阳性对照组以及不同浓度的复方血栓通观察组．每组均加入终浓度为10ng／ml的血管内皮生长因子（VEGF）．阳性对照组加入终浓度0．20mg／ml的硫酸鱼精蛋白，复方血栓通观察组设立0．39～50．00mg／ml的多个不同浓度组。培养24h后采用酶联免疫检测仪测定各组吸光度4值，观察不同浓度的复方血栓通对VEGF诱导的RF／6A细胞增殖的影响。②设立空白对照组和终浓度为3．13、6．25、12．50mg／ml的复方血栓通组，培养24h后检测不同浓度复方血栓通对RF／6A细胞移行的影响。⑧设立空白对照组和终浓度为0．39、0．78、1．56mg／ml的复方血栓通组，培养12h后检测不同浓度的复方血栓通对RF／6A细胞管腔形成的影响；④设立空白对照组、终浓度为0．20mg／ml的硫酸鱼精蛋白阳性对照组和终浓度为1．56、3．13、6．25mg／ml的复方血栓通组，培养24h后，运用Western Blot和实时定量逆转录聚合酶链反应（RT．PCR）的方法，分别检测不同浓度的复方血栓通对RF／6A细胞表达VEGF、基质金属蛋白酶2（MMP．2）蛋白和mRNA的影响。对多组计量资料进行单因素方差分析。结果①各组间4值差异有统计学意义（F=158．669，P〈0．01），同空白对照组比较，0．78～25．00mg／ml浓度的复方血栓通对VEGF诱导的RF／6A细胞增殖具有明显抑制作用（尸均〈0．011；②复方血栓通浓度为0、3．13、6．25和12．50mg／ml时，RF／6A细胞移行数分别为123．3±13．8、114．3±15．5、54．0±6．1和40．3±10．1，组间差异有统计学意义（B36．918，P〈0．01）；与空白对照组比较，6．25和12．50mg／ml浓度的复方血栓通对RF／6A细胞移行具有明显�Objective To investigate the effect and mechanism of Complex Xueshuantong on the angiogenesis of rhesus choroid-retina endothelial （RF/6A） cells. Methods This experimental research included 4 parts. ①RF/6A cells in logarithmic growth phase were cultured and divided into blank control group, positive control group （0.20 mg/ml protamine） and Complex Xueshuantong groups at different concentrations （0.39 to 50.00 mg/ml）. Vascular endothelial growth factor （VEGF） at terminal concentration of 10 ng/ml was added into all groups. The optic absorptions （A values） of each group were measured 24 hours later. ②RF/6A cells in logarithmic growth phase were cultured in Transwell insert and divided into blank control group, Complex Xueshuantong groups at different t concentrations of 3.13, 6.25, 12.50 mg/ml. The number of cells that migrated to under the Transwell membrane was counted after cultured for 24 hours. ③RF/6A ceils were divided into blank control group, Complex Xueshuantong groups at different concentrations of 0.39, 0.78, 1.56 mg/ml. The number of full-formed tubes was counted after cultured for 12 hours. ④RF/6A cells were divided into blank control group, positive control group of protamine at terminal concentration of 0.20 mg/ml, and Complex Xueshuantong groups at different concentrations of 1.56, 3.13, 6.25 mg/ml. Western Blot and real-time quantitative RT-PCR were used to detect the expression of VEGF and matrix metalloproteinase 2 （MMP-2） protein and mRNA of RF/6A ceils after cultured for 24 hours. Data were statistically analyzed by ANOVA method. Results ①The difference of the A values within groups was statistically significant （F=158.669, P〈0.01）. Compared with that of blank control group, the A values of Complex Xueshuantong groups at different concentrations from 0.78 to 25.00 mg/ml were significantly reduced （all P〈0.01）, the proliferation of RF/6A cells induced by VEGF was inhibited by Complex Xueshuantong at certain concentrations. ②When the concentrations

关 键 词：复方血栓通 中草药 血管生成抑制剂 脉络膜新生血管 基质金属蛋白酶类 血管内皮生长因子类 细胞增生 细胞移行 

分 类 号：R276.7[医药卫生—中医五官科学]