原花青素对氧化应激人眼小梁网细胞凋亡及细胞色素C释放的影响

目的 研究原花青素对H2O2诱导人眼小梁网细胞(human trabecular meshwork cells,HTMC)凋亡及细胞色素C释放的影响.方法 将HTMC进行传代后随机分为5组.未处理组:正常培养的HTMC;对照组:正常培养的HTMC+H2O2(500μmol·L-1处理lh);3个浓度的原花青素组:正常培养的HTMC+H2O2(500 μmol· L-1处理lh)+原花青素(浓度分别为0.02 g·L-1、0.05 g·L-1、0.10g·L-1).使用Annexin V-FITC检测细胞凋亡情况,Western blot检测HTMC胞浆细胞色素C含量.结果 与未处理组相比,对照组及各原花青素组细胞凋亡率均升高,差异均有统计学意义(均为P＜0.01).与对照组相比,各原花青素组细胞凋亡率均降低,差异均有统计学意义(均为P＜0.01);不同浓度原花青素组间随原花青素浓度增加细胞凋亡率下降,各原花青素组间差异均有统计学意义(均为P＜0.01).与未处理组相比,对照组以及0.02 g·L-1和0.05 g· L-1原花青素组细胞色素C的释放均增加,差异均有统计学意义(均为P＜0.01);而0.10 g·L-1原花青素组与未处理组相比,差异无统计学意义(P＞0.05).与对照组相比,各原花青素组细胞色素C释放均减少,差异均有统计学意义(均为P＜0.01).不同浓度原花青素组间随原花青素浓度增加细胞色素C的释放减少,各组间差异均有统计学意义(均为P＜0.01).结论 外源性原花青素可以降低氧化应激HTMC的凋亡率,减少细胞色素C的释放,有较强的抗氧化作用,并在一定浓度范围内随着浓度的增加其抗氧化作用逐渐增强.

Effects of procyanidins on cell apoptosis and the release of cytochrome C in human trabecular meshwork cells under oxidative stress

Objective To investigate the effect of procyanidins (PC) on cell apoptosis and cytochrome C releasing in human trabecular meshwork cells (HTMC) under oxidative stress induced by H2 O2.Methods HTMC were cultured and then divided randomly into 5 groups.Normal cultured HTMC served as normal group without treatment,and normal cultured HTMC in control group were treated with 500 μrnol · L-1 H2O2 for 1 h,as well as normal cells in treatment groups were treated with 500 imol · L-1 H2 O2 for 1 h combined with different concentrations of PC (0.02 g · L-1,0.05 g · L-1,0.10 g · L-1).Then flow cytometry and Western blot were applied to detect the cell apoptosis rate and the release of cytochrome C (CytC) respectively.Results Compared with the normal group,the apoptotic rates of the control group and the PC treatment groups were significantly increased,and the difference was statistically significant (all P ＜ 0.01).Compared with the control group,the apoptotic rates of the 3 PC treatment groups were decreased,and the differences were statistically significant (all P ＜ 0.01).Moreover,the apoptotic rate in 0.02 g · L-1,0.05 g · L-1,0.10 g · L-1 PC treatment groups was decreased as PC concentration increased,and the differences were statistically significant (all P ＜ 0.01).Furthermore,compared with the normal group,the release of cytochrome C in the control group,0.02 g · L-1 and 0.05 g · L-1 PC treatment group enhanced,and the differences were statistically significant (all P ＜ 0.01),but there was no significant difference between 0.01 g · L-1 PC treatment group and the normal group (P ＞ 0.05).Compared with the control group,the release of cytochrome C in the 3 PC treatment group was attenuated with significant differences (all P ＜0.01).Meanwhile,the release of cytochrome C was decreased in the 3 PC treatment groups with a concentration-dependent manner,and the pairwise differences were statistically significant (all P ＜ 0.01).Conclusion Exogenous PC can decrease the apoptotic rate of HTMC under oxidative stress and reduce the release of cytochrome C,as well as it also has an antioxidant effect in a certain concentration-dependent manner.