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目的研究不同浓度同型半胱氨酸(Hcy)对静息和钙离子导体(A23187)激活状态下人脐静脉内皮细胞中内皮型一氧化氮合酶(eNOS)和小凹蛋白-1(caveolin-1)mRNA和蛋白表达的影响。方法人脐静脉内皮细胞(HU-VECs)随机分4组:对照组、Hcy组(20、50、100、300μmol/L)、A23187(1μmol/L)组及上述浓度Hcy分别与A23187(1μmol/L)共同孵育组,测定各组eNOS和caveolin-1蛋白及mRNA表达情况,同时检测各组一氧化氮(NO)、丙二醛(MDA)含量和eNOS、超氧化物歧化酶(SOD)活力。结果对照组caveolin-1蛋白表达为1.13,Hcy各组该蛋白为0.21～2.05,呈浓度依赖性增高,差异有统计学意义(F=30.163,P<0.05);各组eNOS mRNA表达为1.80～2.08,差异无统计学意义;对照组eNOS活力和NO含量分别为1.20 U/mL和122.41μmol/L,Hcy组eNOS活力和NO含量分别为0.65～0.74 U/mL和65.33～98.91μmol/L,均呈浓度依赖性降低,差异均有统计学意义(F=5.12、18.91,P<0.05)。结论 Hcy可能通过增强caveolin-1蛋白表达,使其与eNOS藕联增加,进而抑制了eNOS活力。
Objective To investigate the effects of different concentrations of homocysteine (Hcy) on the activity of endothelial nitric oxide synthase (eNOS) and caveolin-1 in human umbilical vein endothelial cells (HUVECs) activated by resting and calcium ion conductor (A23187) 1) mRNA and protein expression. Methods Human umbilical vein endothelial cells (HU-VECs) were randomly divided into 4 groups: control group, Hcy group (20,50,100,300μmol / L), A23187 (1μmol / L). The mRNA and protein expressions of eNOS and caveolin-1 in each group were measured. The levels of nitric oxide (NO), malondialdehyde (MDA), eNOS and superoxide dismutase (SOD) . Results The expression of caveolin-1 in control group was 1.13 and the protein in Hcy group was 0.21-2.05, showing a significant increase in concentration-dependent manner (F = 30.163, P <0.05). The eNOS mRNA expression in each group was 1.80 ~ 2.08, the difference was not statistically significant. The eNOS activity and NO content in control group were 1.20 U / mL and 122.41μmol / L, respectively. The eNOS activity and NO content in Hcy group were 0.65-0.74 U / mL and 65.33-98.91μmol / L, All in a concentration-dependent manner, the differences were statistically significant (F = 5.12,18.91, P <0.05). Conclusions Hcy may increase the expression of caveolin-1 and increase its coupling with eNOS, thus inhibiting eNOS activity.