白藜芦醇对过氧化氢诱导人动脉平滑肌细胞的影响及其机制

目的：探讨白藜芦醇（Res）对过氧化氢（H2O2）诱导的人动脉平滑肌细胞的影响及其可能机制。方法：在人动脉平滑肌细胞中加入不同浓度（20、100、500umol／L）H2O2处理24h，建立动脉平滑肌细胞氧化损伤模型。不同终浓度（2、10、50umol／L）的Res单纯作用或分别与100umol／LH202共同作用于动脉平滑肌细胞24h。应用MTT比色法检测细胞存活率，黄嘌呤氧化酶法检测超氧化物歧化酶（SODl的活力，硫代巴比妥酸比色法检测丙二醛（MDA）含量，2，7-二氯荧光黄双乙酸盐（DCFH—DA）染色荧光显微镜照相检测细胞内的活性氧簇（ROS）水平，流式细胞术检测细胞凋亡百分率。结果：20、100、500umol／LH2O2处理人动脉平滑肌细胞24h后，动脉平滑肌细胞存活率和培养基上清液SOD活力均呈剂量依赖性地下降，细胞蛋白裂解液MDA含量活性则呈剂量依赖性地升高（P〈0．05）。2、10和50txmol／LRes与100umol／LH2O2共同作用动脉平滑肌细胞24h后，可呈剂量依赖性地抑制H2O2诱导的细胞存活率、SOD活力的下降以及MDA含量的升高，其中10和50umol／LRes＋100umol／LH202处理组与100umol／LH2O2模型组比较，差异均有统计学意义（均P〈0．05）。10和50umol／LRes＋100Ixmol／LH202处理组DCF荧光强度均显著低于100txmol／LH2O2模型组（P均〈0．05）。10和50btmol／LRes＋100umol／LH2O2处理组人动脉平滑肌细胞的凋亡率分别为（20．7±3．0）％和（13．2±1．5）％，均显著低于100umol／LH2O2模型组的（27．2±3．6）％（均P〈0．05）。结论：Res可对抗H2O2诱导的动脉平滑肌细胞氧化损伤，其机制可能与其减少ROS生成以及降低动脉平滑肌细胞凋亡率有关。

Effects and mechanisms of resveratrol on hydrogen peroxide-induced human arterial smooth muscle cells

t Objective ： To investigate the effects and mechanisms of resveratrol （Res） on hydrogen peroxide （H2 02 ）-induced human arterial smooth muscle cells. Methods： Human arterial smooth muscle cells were incu- bated with H202 （20, 100 and 500umol/L） for 24h to establish the oxidative stress-induced cell injury model. This was followed by the addition of different concentrations of Res （2, 10 and 50umol/L） with or without H202 （100umol/L） to the arterial smooth muscle cells for 24h. The viability of human arterial smooth muscle cells was measured by MTF assay, and the SOD activity was assayed by using xanthine oxidase method. Thiobarbituric acid eolorimetrie approach was employed to determine the level of MDA. The level of reactive oxygen species （ROS） was detected by DCFH-DA staining and photofluorography, and the percentage of apoptotic cells was as- sessed by using flow eytometry （FCM）. Results： Following treatment with 20, 100 and 500pumol/L H2 02 for 24 h, there was a reduction in the cell viability and SOD activity in the supernatant whilst increased level of MDA, in a concentration-dependent manner, in the human arterial smooth muscle cell supernatant （all P 〈0.05）. In- cubation with 2, 10 and 50p.mol/L Res anO 100umol/L H202 for 24 h ameliorated the reduced cell vitality, SOD activity and the level of MDA in a dose-dependent fashion. The differences in SOD activity differed substantially between 100umol/L H202 model group and 10 and 50 umol/L Res plus 100umol/L H202 treatment groups （both P 〈0.05）. Compared with 100 umol/L H2O2 model group, the DCF fluorescence intensity was signifi-cantly increased in 10 and 50umol/L Res plus 100 umol/L H202 treatment groups （ both P 〈0.05 ）. The 10 and 50umol/L plus 1001umol/L H202 treatment groups were associated with reduced apoptosis rate compared with 100 umol/L H2 02 model group （20.7 ± 3.0） % and （ 13.2 ± 1.5 ） % vs. （27.2 ± 3.6） % ]. Conclusion ： Res protected human arterial smooth muscle cells against H2Oz-induced oxidative stress injury, presumably associated with redueed production of Res and apoptosis rate of vascular smooth muscle cells.