补阳还五汤对缺氧—复氧内皮细胞ICAM—1 mRNA表达影响的研究

The Effects of Buyang Huanwu Decoction on the Expression of ICAM-1 mRNA in the Endothelial Cells Cultured in Hypoxic-Reoxgenative Circumstance [BHDWG0.1mm,WK40W]

目的探讨补阳还五汤对缺氧 — 复氧内皮细胞ICAM—1mRNA表达的影响及其保护作用。方法取内皮细胞株EVC30 4 ,随机分为 5组进行相应处理。空白组 (A) 37℃ ,5 %CO2 正常培养 2 4h ;缺氧组 (B)持续通入95 %N2 +5 %CO2 混合气 2 4h ;缺氧 — 复氧组 (C)持续通入 95 %N2 +5 %CO2 混合气 12h后 ,改用正常培养 12h ;西药组 (D)实验前加入终浓度为 10ng L的尼莫地平 ,方法同C组 ;补阳还五汤组 (E) :以补阳还五汤培养 ,方法同C组。硫氰酸胍 — 酚 — 氯仿法提取各组细胞总RNA ,以 β— Actin为内对照 ,将RT—PCR产物用 1.5 %的琼脂糖凝胶电泳 ,用ICAM—1和β—Actin的光密度比值表示mRNA水平。结果缺氧 — 复氧组ICAM—1mRNA表达最高 (P <0 .0 1) ,补阳还五汤组和西药组均显示ICAM—1mRNA表达抑制 (P <0 .0 1,P <0 .0 1)。结论补阳还五汤能抑制内皮细胞ICAM—1mRNA异常表达 ,对缺氧 — 复氧内皮细胞起保护作用。

Objective To investigate the effects of Buyang Huwan Decoction(BHD) on the expression of intercellular adhesion molecule-1 (ICAM-1) mRNA in the endothelial cell line EVC304 cultured in hypoxic-reoxgenative circumstance. Methods The cuturled cells of ECV304 were randomly divided into 5 groups, i.e. blank group (A), hypoxic group (B), hypoxic-reoxgenative group (C),drug controlling group (D) and BHD group (E), all being treated by corresponding conditions respectively. Then, the expression of ICAM-1 mRNA was determined by reverse transcriptase polymerase chain reaction (RT-PCR). Results The highest level of ICAM-1 mRNA was determined in group C (P<0.01). In contrast,the expressive inhibition of ICAM-1mRNA was tested in both BHD group and group D.Conclusions BHD can inhibit ICAM-1 mRNA transcription of endothelial cells in the circumstance of hypoxic-reoxgenation. In this way, it is effective in the protection of endothelial cells in the circumstance of hypoxic-reoxgenation.