黄酒对TNF-α诱导的大鼠血管内皮细胞功能及动脉粥样硬化的影响

Effects of Chinese yellow wine on rat vascular endothelial cells and atherosclerosis induced by TNF-α

目的：观测黄酒对 TNF －α诱导的大鼠血管内皮细胞的作用，分析其对动脉粥样硬化的影响。方法大鼠原代主动脉 VECs 经分离培养及纯化鉴定后，取第3～4代细胞用于实验。根据前期实验，分为对照组、TNF －α组、TNF －α＋瑞舒伐他汀组（10μmol／L）、TNF －α＋酒精组（0．5％、1．0％、1．5％）、TNF －α＋黄酒组（0．5％、1．0％、1．5％），共有5种不同处理的9个水平组。培养24 h 后收集样品，硝酸还原酶法测定培养液上清液 NO 的含量；培养48 h 后用免疫印迹法检测 eNOS 和 iNOS 蛋白的表达量。结果与 TNF －α组相比，瑞舒伐他汀组、黄酒1．0％组、黄酒1．5％组内皮上清中 NO 含量升高，eNOS 活性增加，eNOS 蛋白的表达升高， iNOS 蛋白表达降低（P ＜0．05）；与瑞舒伐他汀组相比，黄酒1．0％组和黄酒1．5％组的 eNOS 蛋白表达降低， iNOS 蛋白表达升高（P ＜0．05）；相比酒精组（1．0％或1．5％），其对应酒精度的黄酒中 eNOS 蛋白表达升高（P ＜0．05），iNOS 蛋白表达降低（P ＜0．05）。结论小剂量黄酒能够使上清 NO 含量增加，增强 eNOS 的活性及其蛋白表达，抑制 iNOS 蛋白表达，具有类他汀样作用，这可能是其抗动脉粥样硬化的机制之一。

Objective To determine similarities in effect of yellow wine as compared statin and the possibility that yellow wine inhibits TNF -α-induced NO production,eNOS activity,expression of eNOS and iNOS protein in cultured rat VECs.Methods Isolation,cultivation,purification and identification of vascular endothelial cells of rat thoracic aorta in vitro were conducted.The passages 3 or 4 of VECs were used in all studies.Then we divided cells into 9 groups according to assays before：control,TNF -α,TNF -α＋rosuvastatin （10 umol/L）,TNF -α＋ethanol 0.5%,TNF -α＋yellow wine 0.5%,TNF -α＋ethanol 1.0%,TNF -α＋yellow wine 1.0%,TNF -α＋ethanol 1.5%,and TNF -α＋yellow wine 1.5% and the cells were given the corresponding treatment.NO production of culture supernatant was determined by nitrate reduction method and eNOS activity of cells was measured by chemical colorimetric method after the corresponding treatment for 24 h.The expression of eNOS and iNOS protein were detected by western blotting after the corresponding treatment for 48 h.Results Compared with the TNF -αgroup,NO production,eNOS activity,and eNOS protein expression in the rosuvastatin,and yellow wine 1.0%,and 1.5% groups were significantly increased and expression of iNOS protein were significantly decreased.Compared with the rosuvastatin group,eNOS protein expression in yellow wine 0.5% and yellow wine 0.5% groups significantly decreased.The expression of iNOS protein were significantly increased. Compared with ethanol groups,eNOS protein expression in yellow wine 0.5% and yellow wine 0.5% groups significantly increased and expression of iNOS protein were significantly decreased.Conclusion Treatment with yellow wine increased NO production,eNOS activity,and eNOS protein expression,which decreases iNOS protein expression.We conclude that yellow wine has similar beneficial effects as rosuvastatin on the cardiovascular system.These effects may be attributed to their anti -atherosclerotic actions.