补肾养血明目方对ARPE-19细胞氧化损伤的保护机制研究

Study on protective mechanisms of Bushen Yangxue Mingmu Formula on oxidative injury of ARPE-19 cells

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 oxidative 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 superoxide 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 levels,decreased the production of ROS and alleviated mt DNA 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.