雷公藤甲素对人视网膜色素上皮细胞氧化损伤的抑制作用

Inhibition effects of triptolide on human retinal pigment epithelium cells against oxidative damage

目的探讨雷公藤甲素对人视网膜色素上皮(retinal pigment epithelium,RPE)细胞氧化损伤的抑制作用及其机制。方法RPE细胞传代培养,分为对照组、氧化损伤组(100μmol·L^(-1)H_2O_2)和雷公藤甲素(100μmol·L^(-1)、200μmol·L^(-1))干预组。应用MTT比色法检测细胞增殖率,流式细胞技术测定细胞内活性氧(reactive oxygen species,ROS)含量,Hoechst33258染色观察细胞凋亡,Western-blot检测RPE细胞内超氧化物歧化酶(superoxide dismutase,SOD)及丙二醛(malondialdehyde,MDA)蛋白的表达水平。结果雷公藤甲素能明显抑制H_2O_2诱导的RPE细胞活性的下降,用不同浓度雷公藤甲素处理后,RPE细胞存活率分别提高到(56. 00±2. 76)%和(70. 33±3. 85)%,与氧化损伤组[(32. 67±3. 08)%]相比差异均有统计学意义(均为P <0. 05)。雷公藤甲素能减少细胞内ROS生成,抑制细胞凋亡,不同浓度雷公藤甲素处理后,RPE细胞凋亡率分别下降到(50. 33±4. 61)%和(46. 67±4. 73)%,与氧化损伤组(67. 00%±3. 42%)相比差异均有统计学意义(均为P <0. 05)。雷公藤甲素组SOD蛋白表达显著高于氧化损伤组,MDA蛋白表达显著低于氧化损伤组(P <0. 05)。结论雷公藤甲素对RPE细胞氧化损伤具有抑制作用,其作用机制与上调SOD的表达及下调MDA的表达有关;雷公藤甲素有望成为治疗视网膜病变的有效药物。

Objective To discuss the inhibition effect and mechanism of triptolide on retinal pigment epithelium(RPE)cells induced by oxidative damage.Methods RPE cells were subcultured and divided into control group,oxidative damage group(100μmol·L^-1 H2O2)and triptolide(100μmol·L^-1,200μmol·L^-1)intervention group.MTT colorimetry was used to detect cells proliferation rate and flow cytometry technology was used to determine the expression of reactive oxygen species(ROS)within cells,Hoechst33258 staining was used to observe nucleus apoptosis,the protein expression of superoxide dismutase(SOD)and malondialdehyde(MDA)within cells were detected by Western-blot.Results Triptolide could significantly inhibit RPE cells activity decrease induced by H2O2,after treated with different concentrations off triptolide,RPE cell activity increased to 56.00%±2.67%and 70.33%±3.85%respectively,compared with the oxidative damage group[(32.67±3.08)%],the differences have statistically significant(P<0.05);Triptolide could reduce ROS generation within cells and inhibit cells apoptosis,after treated with different concentrations of triptolide,RPE cell apoptosis rate decreased to(50.33±4.61)%and(46.67±4.73)%respectively,compared with the oxidative damage group(67.00±3.42)%,the differences have statistically significant(P<0.05);The protein expression of SOD in triptolide group were significantly higher than that in oxidative damage group,the protein expression of MDA were significantly lower than that in oxidative damage group(P<0.05).Conclusion Triptolide has inhibition effect in RPE cells oxidative damage,which related to raise expression of SOD and suppress expression of MDA,triptolide expected to become an effective drug for treatment of retinopathy.