摘要
目的：探讨体外葡萄籽原花青素对氯化钴致视网膜神经节细胞RGC－5缺氧损伤模型的保护作用。方法：采用氯化钴（400μmol／L）诱导RGC－5细胞化学缺氧损伤，观察葡萄籽原花青素（40μg／mL ）对缺氧的抗损伤作用。以MTT法检测细胞存活率；Hoechst 33342荧光染色法观察细胞凋亡情况；以H2 DCFDA荧光法检测细胞内活性氧的变化；实时定量PCR检测细胞Bcl－2、半胱酰胺天冬氨酸蛋白酶（ caspase ）9和caspase 3 mRNA水平。结果：与缺氧对照组比较，葡萄籽原花青素预处理可提高细胞存活率（ P＜0．001）、减少细胞凋亡（ P＜0．001） ，并减少细胞内的活性氧（ P ＜0．001）；葡萄籽原花青素还显著升高 Bcl－2基因的表达（ P ＜0．001） ，显著降低caspase 9和caspase 3基因的表达（ P＜0．001）。结论：葡萄籽原花青素对氯化钴诱导的视网膜神经节RGC－5细胞缺氧损伤起保护作用，其保护作用可能与细胞内活性氧减少、Bcl－2基因表达上调及casepase 9和casepase 3基因表达下调有关。
Objective： To investigate the protective effects of grape seed proanthocyanidin extracts （ GSPE ） against CoCl 2-induced hypoxic injury in cultured RGC-5 cells.Mtehods： CoCl 2（ 400 μmol L/） was used to induce hypoxic injury in cultured RGC-5 cells;the cells were pretreated with 0, 100, 200, 400 and 800μmol/L GSPE for 24h. The cell viability was assayed by MTT; the apoptosis was detected by Hoechst 33342 staining;the intracellular reactive oxygen species （ ROS） was measured by H2DCFDA oxidative reaction.The mRNA expression of Bcl-2, caspase 9 and caspase 3 was determined by real-time PCR.Results： Compared to hypoxic control group, pretreatment with GSPE significantly increased viability of RGC-5 cells （ P 〈0.001 ） , reduced cell apoptosis （ P〈0.001 ） and intracellular ROS （ P〈0.001 ） .In addition , GSPE significantly increased the mRNA expression of Bcl-2 （P〈0.001 ） and decreased mRNA expression of caspase 9 （ P 〈0.001 ） and caspase 3 （ P〈0.001 ） compared to hypoxic control group .Conclusion： GSPE may have a protective effect against CoCl 2-induced hypoxic injury in cultured RGC -5 cells. The decrease of intercellular ROS , up-regulation of Bcl-2 and down-regulation of caspase 9 and caspase 3 may be involved in the mechanism of the protective effect of GSPE .
出处
《浙江大学学报（医学版）》
CAS
CSCD
北大核心
2015年第1期24-29,共6页
Journal of Zhejiang University(Medical Sciences)
基金
浙江省教育厅科学研究项目(Y201328427)
浙江省中西医结合学会科研基金(2012LY013)
