补肾养血明目方对ARPE-19细胞氧化损伤的保护机制研究

摘要

目的 探讨补肾养血明目方对氢醌(hydroquinone,HQ)诱导的人视网膜色素上皮细胞(ARPE-19)氧化损伤的保护机制.方法 采用HQ制作ARPE-19细胞氧化应激损伤模型,分为正常对照组,模型组,空白肠吸收液组,预防给药组和治疗给药组.分别采用TUNEL技术观察细胞凋亡情况,免疫组化技术检测8-羟基脱氧鸟苷(8-OHdG)的表达情况,化学发光法测定细胞内ATP含量,荧光法测定活性氧(ROS)水平,化学比色法检测细胞中超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)含量.结果 与氧化损伤模型组相比,补肾养血明目方可减少ARPE-19细胞凋亡,抑制细胞内ATP的水平下降,减少ROS的生成,减少RPE细胞8-OHdG阳性表达率,提高抗氧化酶(SOD和GSH-Px)的含量.结论 线粒体保护是补肾养血明目方发挥抗HQ诱导的ARPE-19细胞氧化损伤的重要途径.%OBJECTIVE To investigate the possible protective mechanism of Bushen Yangxue Mingmu (BSYXMM) Formula on ARPE-19 cells under oxidative stress induced by Hydroquinone (HQ). METHODS The ox-idative stress models of ARPE-19 cells were induced by HQ and divided into normal control group, model group, blank group, preventive group and treatment group. The percentage of apoptotic cells was measured by TUNEL assay and the expression of 8-OHdG was detected by immunohistochemical analysis. Chemiluminescence detection was used to determine ATP (adenosinetriphosphate) and ROS (reactive oxygen species) content. The contents of superox-ide dismutase (SOD) and glutathione peroxidase (GSH-Px) were detected by colorimetry. RESULTS Compared to the model group, BSYXMM Formula significantly reduced apotosis in ARPE-19 cells, inhibited the decline of ATP lev-els, decreased the production of ROS and alleviated mtDNA damage. Moreover, BSYXMM Formula increased the content of SOD and GSH-Px. CONCLUSIONS Mitochondrial protection was the important pathway for BSYXMM formula protecting against HQ-induced oxidative stress injury in ARPE-19 cells.