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

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

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+H_2O_2(500μmol·L^(-1)处理1 h);3个浓度的原花青素组:正常培养的HTMC+H_2O_2(500μmol·L^(-1)处理1 h)+原花青素(浓度分别为0.02 g·L^(-1)、0.05 g·L^(-1)、0.10 g·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。

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 H2O2.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 μmol·L-1 H2O2 for 1 h,as well as normal cells in treatment groups were treated with 500 μmol·L-1 H2O2 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.