目的:探讨补肾养血明目方含药肠吸收液对氢醌(hydroquinone,HQ)诱导的人视网膜色素上皮细胞(ARPE-19)氧化应激损伤的保护作用。方法:采用不同浓度HQ诱导体外培养ARPE-19细胞氧化损伤,确定最佳造模浓度;制备补肾养血明目方含药肠吸收液。采用CCK-8法检测补肾养血明目方含药肠吸收液对ARPE-19细胞活力的影响,TUNEL技术观察其对细胞凋亡的保护作用,以及化学比色法检测细胞中超氧化物歧化酶(superoxide dismutase,SOD)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)的活性。结果:当HQ浓度达到90μmol/L以上时,ARPE-19细胞活性明显降低。相较于模型组,预防给药时,浓度1%和2%补肾养血明目方含药肠吸收液对HQ损伤具有有显著保护作用(均P〈0.01),0.5%和5%含药肠吸收液对HQ损伤有一定保护作用(均P〈0.05);治疗给药时,1%和2%含药肠吸收液对HQ损伤有一定保护作用(均P〈0.05)。与模型组相比,预防组的细胞凋亡率显著下降(P〈0.01),治疗组的细胞凋亡率有一定下降(P〈0.05)。抗氧化酶检测结果显示与模型组相比,预防组和治疗组均能明显提高SOD和GSH-Px含量(均P〈0.05),其中预防组的作用更为显著(P〈0.01,P〈0.05)。结论:补肾养血明目方对HQ诱导的ARPE-19细胞氧化损伤有保护作用,其作用机制可能与增加细胞内抗氧化酶的含量有关。
AIM: To explore the effect of Bu Shen Yang Xue Ming Mu (BSYXMM) Formula on hydroquinone-induced oxidative stress injury in ARPE-19 cells.METHODS: The oxidative injury model of ARPE-19 cell was induced by exposure to various concentrations of hydroquinone (HQ) to determine the optimal concentration.Intestinal absorption solutions of BSYXMM Formula were prepared.Effect of intestinal absorption solutions of BSYXMM Formula on the cell viability was detected by CCK-8 assay,and the percentage of apoptotic cells was measured by TUNEL assay.The levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in ARPE-19 cells were detected by means of chemical colorimetry.RESULTS: It was found that ARPE-19 cell viability significantly decreased when the concentration of HQ was higher than 90μmol/L.Compared with the model group,1% and 2% intestinal absorption solutions in the pre-treatment group could significantly alleviate HQ-induced injury (P〈0.01) and 0.5% and 5% intestinal absorption solutions in the pre-treatment group could alleviate the injury in certain degree(P〈0.05).While in the treatment group 1% and 2% intestinal absorption solutions could alleviate the injury to some extent (P〈0.05).TUNEL results showed that the apoptosis rate decreased significantly in the pre-treatment group (P〈0.01)and to some extent in the treatment group (P〈0.05)compared with the model group.It was shown that both levels of SOD and GSH-Px in pre-treatment group and treatment group were markedly higher than that of model group(P〈0.05),and pre-treatment group had more significant effect (P〈0.01,P〈0.05).CONCLUSION: BSYXMM Formula could protect against HQ-induced oxidative stress injury in ARPE-19 cells,which may be related with the increasing of antioxidant enzyme in the cells.