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目的:探讨白藜芦醇(Res)对过氧化氢(H_2O_2)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、500μmoL/L)H_2O_2处理24 h,建立动脉平滑肌细胞氧化损伤模型。不同终浓度(2、10、50μmoL/L)的Res单纯作用或分别与100μmol/L H_2O_2共同作用于动脉平滑肌细胞24 h。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(SOD)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、500μmol/L H_2O_2处理人动脉平滑肌细胞24 h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50μmol/L Res与100μmol/L H_2O_2共同作用动脉平滑肌细胞24 h后.可呈剂量依赖性地抑制H_2O_2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50μmol/L Res+100μmoL/L H_2O_2,处理组与100μmol/L H_2O_2,模型组比较,差异均有统计学意义(均P<0.05)。10和50μmol/L Res+100μmol/L H_2O_2处理组DCF荧光强度均显著低于100μmoL/L H_2O_2,模型组(P均<0.05)。10和50μmoL/L Res+100μmoL/L H_2O_2,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100μmol/L H_2O_2,模型组的(27.2±3.6)%(均P<0.05)。结论:Res可对抗H_2O_2诱导的动脉平滑肌细胞氧化损伤.其机制可能与其减少R0s生成以及降低动脉平滑肌细胞凋亡率有关。
Aims: To investigate the effect of resveratrol on human arterial smooth muscle cells induced by hydrogen peroxide (H2O2) and its possible mechanism. Methods: H22 cells were treated with H 2 O 2 at different concentrations (20, 100, 500μmoL / L) for 24 h to establish a model of arterial smooth muscle cell oxidative injury. Effects of different concentrations of Res (2, 10, 50μmoL / L) alone or in combination with 100μmol / L H 2 O 2 on arterial smooth muscle cells for 24 h. Cell viability was measured by MTT colorimetric assay. The activity of superoxide dismutase (SOD) was detected by xanthine oxidase method. The content of malondialdehyde (MDA) was detected by thiobarbituric acid colorimetry. The level of reactive oxygen species (ROS) in the cells was detected by DCFH-DA staining and fluorescence microscope. The percentage of apoptosis was detected by flow cytometry. RESULTS: After treated with 20, 100, 500μmol / L H 2 O 2 for 24 h, the viability of arterial smooth muscle cells and the activity of superoxide dismutase (SOD) in culture supernatant decreased in a dose-dependent manner. The activity of MDA in cell lysate Dependently increased (P <0.05). 2, 10 and 50μmol / L Res combined with 100μmol / L H 2 O 2 for 24 h could inhibit the cell viability induced by H 2 O 2, the decrease of SOD activity and the increase of MDA content in a dose-dependent manner 50μmol / L Res + 100μmoL / L H 2 O 2, the treatment group and 100μmol / L H 2 O 2, the model group, the difference was statistically significant (P <0.05). The fluorescence intensity of DCF in 10 and 50μmol / L Res + 100μmol / L H 2 O 2 treatment group was significantly lower than that in 100μmoL / L H 2 O 2 group (all P <0.05). The apoptotic rates of smooth muscle cells of human arteries were (20.7 ± 3.0)% and (13.2 ± 1.5)%, respectively, which were significantly lower than those of 100 μmol / L H 2 O 2 in 10 and 50μmoL / L Res + 100μmoL / (27.2 ± 3.6)% (all P <0.05). CONCLUSION: Res can counteract H 2 O 2 -induced oxidative damage of arterial smooth muscle cells, which may be related to the decrease of R0s production and the decrease of apoptosis rate of arterial smooth muscle cells.