黄酒对同型半胱氨酸诱导的大鼠血管内皮细胞基质金属蛋白酶2表达的影响

 ［摘要］目的:观察黄酒和红葡萄酒是否能抑制同型半胱氨酸(Hcy)诱导的大鼠血管内皮细胞(VECs)中基质金属蛋白酶2(MMP-2)的表达。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。将Hcy(0、50、100、500、1 000 μmol/L)与VECs共同孵育48 h及100 μmol/L Hcy与VECs共同培养24、48、72 h，分别用免疫印迹和明胶酶谱法检测Hcy 对MMP-2蛋白表达和活性的影响，确定Hcy最佳干预的浓度和时间。每种酒（1.0%、1.2%、1.4%、1.6%、1.8%、2.0%）与100 μmol/L Hcy共同孵育VECs 48 h， MTT法检测酒类对细胞活性的影响，确定最佳干预浓度后处理分control组、Hcy组、Hcy+黄酒组、Hcy+红酒组和Hcy+酒精组5组。培养48 h后收集样品，免疫印迹法检测VECs中MMP-2的表达量，明胶酶谱法测定VECs上清液中MMP-2的活性。结果:Hcy在50~500 μmol/L呈浓度依赖性地促进MMP-2蛋白表达和活性的增强（P<0.05或P<0.01），Hcy在100 μmol/L时干预24、48、72 h MMP-2蛋白表达和活性的增强呈时间依赖性，48 h时最明显（P<0.01）。与Hcy组相比，黄酒组和红酒组VECs MMP-2蛋白表达及上清液中MMP-2活性显著下降（P<0.05），酒精组MMP-2蛋白表达及上清液中MMP-2活性下降，但差异均无统计学意义（P>0.05）。与酒精组相比，黄酒组和红酒组VECs MMP-2表达及上清液中MMP-2活性的差异有统计学意义（P<0.05），黄酒组和红酒组之间无显著差异（P>0.05）。结论: Hcy能增强血管内皮细胞MMP-2蛋白表达及酶的活性，小剂量的黄酒和红葡萄酒均能抑制Hcy 诱导的MMP-2表达及活化。

Effects of Chinese rice wine on expression of matrix metalloproteinase-2 induced by homocysteine in rat vascular endothelial cells

［ABSTRACT］AIM: To study the possibility that rice wine and red wine inhibit the production of homocysteine (Hcy)-induced matrix metalloproteinase-2(MMP-2) in cultured rat vascular endothelial cells (VECs). METHODS: Isolation, cultivation, purification and identification of VECs of rat thoracic aorta in vitro were conducted. The VECs in passages 3 and 4 were used in all studies. The effects of Hcy at different concentrations and for different durations on the protein expression and activity of MMP-2 were observed by Western blotting and gelatin zymography. The VECs were incubated with one kind of wine (at the concentrations of 1.0%, 1.2%, 1.4%, 1.6%, 1.8% and 2.0%) and 100 μmol/L Hcy for 48 h. The optimal concentration of the wine was selected. In another experiment, the cells were divided into 5 groups: control, Hcy, Hcy+rice wine, Hcy+red wine and Hcy+ethanol, and were given the corresponding treatment for 48 h. The protein level of MMP-2 was detected by Western blotting. The activity of MMP-2 was assayed by gelatin zymography. RESULTS: Hcy at the concentrations of 50~500 μmol/L significantly increased the protein level and activity of MMP-2 in a dose-dependent manner. The cells exposed to Hcy for 24 h, 48 h and 72 h significantly increased the protein expression and activity of MMP-2 in a time-dependent manner. Compared with Hcy group, the protein expression and activity of MMP-2 in rice wine group and red wine group were significantly decreased. No significant difference was shown between ethanol group and Hcy group. Compared with ethanol group, the cells exposed to rice wine and red wine also significantly reduced the protein expression and activity of MMP-2. However, no significant difference between rice wine and red wine groups was found. CONCLUSION: Hcy promotes the protein expression and activity of MMP-2, which may play an important role in pathogenesis of atherosclerosis. Treatment with rice wine or red wine decreases Hcy-induced MMP-2 production in VECs. The attenuation of MMP-2 activation by rice wine and red wine might contribute to their beneficial effects on the cardiovascular system.