杞黄颗粒对H2O2诱导的人视网膜色素上皮细胞炎症损伤的保护作用

目的　探讨杞黄颗粒（QHG）对H₂O₂诱导的人视网膜色素上皮（RPE）细胞系ARPE-19炎症损伤的保护作用。方法　常规培养ARPE-19细胞，随机分为对照组、H₂O₂损伤组(200 μmol?L^-1 H₂O₂处理细胞）、QHG低剂量组（1 g?L^-1 QHG作用24 h后再加入H₂O₂处理细胞）和QHG高剂量组（2 g?L^-1QHG作用24 h后再加入H₂O₂处理细胞）。使用相差显微镜观察各组ARPE-19细胞形态；流式细胞仪检测各组ARPE-19细胞凋亡率，Real-time PCR检测各组ARPE-19细胞β淀粉样蛋白（Aβ)mRNA表达，ELISA检测各组ARPE-19细胞Aβ以及攻膜复合物（MAC）的蛋白表达水平。结果　相差显微镜下可见，QHG低剂量组、QHG高剂量组ARPE-19细胞形态改变均较H₂O₂损伤组轻。对照组、H₂O₂损伤组、QHG低剂量组、QHG高剂量组ARPE-19细胞凋亡率分别为（6.85±0.14）%、（43.60±1.80）%、（23.23±1.43）%、（17.90±0.78）%，H₂O₂损伤组细胞凋亡率较对照组明显增加，QHG低剂量组及QHG高剂量组细胞凋亡率均较H₂O₂损伤组明显降低，且QHG高剂量组较QHG低剂量组降低更明显，差异均有统计学意义（均为P<0.05）。与对照组相比，H₂O₂损伤组ARPE-19细胞Aβ mRNA及Aβ、MAC蛋白表达水平均升高，差异均有统计学意义（均为P<0.01）。与H₂O₂损伤组相比，QHG低剂量组及QHG高剂量组均可明显降低ARPE-19细胞Aβ mRNA 相对表达量及Aβ、MAC蛋白的表达水平，差异均有统计学意义（均为P<0.01）。结论　QHG能有效抑制H₂O₂诱导的人RPE细胞凋亡，减少细胞Aβ及MAC表达，从而起到保护人RPE细胞的作用。

Qihuang Granule protects retinal pigment epithelium cells against H2O2-induced inflammatory injury

Objective To investigate the protective effects of Qihuang granule(QHG)on H2O2-induced inflammatory damage in retinal pigment epithelium(RPE)cell ARPE-19.Methods RPE cells were cultured routinely,and then divided into control group,H2O2 injury group(cells were treated with 200μmol·L-1 H2O2),including QHG low-dose group(cells were treated with 1 g·L-1 QHG for 24 hours and then H2O2 was added)and QHG high-dose group(cells were treated with 2 g·L-1 QHG for 24 hours and then H2O2 was added).The morphological changes,apoptosis rate,mRNA and protein levels of Amyloid-β(Aβ)and membrane attack complex(MAC)in RPE cells were observed by inverted microscope,flow cytometry,RT-PCR and ELISA,respectively.Results It could be seen under a phase contrast microscope that the changes in the QHG low-dose group and QHG high-dose group were more obvious.The apoptosis rates of the control group,H2O2 injury group,QHG low-dose group,and QHG high-dose group were(6.85±0.14)%,(43.60±1.80)%,(23.23±1.43)%,(17.90±0.78)%,respectively.The apoptosis rate of the H2O2 injury group was significantly higher than that of the control group.The apoptosis rate of the QHG low-dose group and the QHG high-dose group was significantly lower than that of the H2O2 injury group,and the QHG high-dose group decreased more significantly than the QHG low-dose group,and the differences were statistical significance(all P<0.05).The expression of AβmRNA and extracellular Aβand MAC protein levels were increased in H2O2 injured group compared with the control group(all P<0.01).QHG low-dose group and high-dose group could significantly improve RPE cell morphology,inhibit apoptosis,and reduce the expression of AβmRNA and extracellular Aβand MAC protein expression compared with H2O2 injured group(all P<0.01).Conclusion QHG can effectively inhibit the apoptosis of RPE cells induced by H2O2,and down-regulate the expression of Aβand MAC,which can protect RPE cells.