白藜芦醇对碘酸钠诱导后人视网膜色素上皮细胞凋亡的抑制作用

Inhibitory effect of resveratrol on apoptosis of human retinal pigment epithelial cells induced by sodium iodate

目的研究白藜芦醇(resveratrol,RES)对碘酸钠诱导的人视网膜色素上皮(retinal pigment epithelial,RPE)细胞凋亡的抑制作用及机制,探索该药是否可用于干性年龄相关性黄斑变性(age-related macular degeneration,AMD)的治疗。方法通过碘酸钠诱导的RPE细胞模拟干性AMD。RPE细胞传代培养分为空白对照组、碘酸钠损伤组、RES+碘酸钠损伤组,并观察各组细胞形态。MTT法检测RES对RPE细胞最佳保护作用的浓度以及最佳浓度下对不同浓度碘酸钠损伤的RPE细胞的保护作用。Western blot检测各组细胞的凋亡蛋白Bax和Bcl-2的表达情况。Hosste33342及流式细胞术检测各组细胞的凋亡总量变化以及凋亡种类之间的差别。结果 5μmol·L^-1 RES对RPE细胞有较强的保护作用且此浓度下对不同损伤程度的RPE细胞都有较强的保护作用。RES+碘酸钠损伤组Bax蛋白的相对表达量(0.45±0.07)明显低于碘酸钠损伤组(0.90±0.09),差异有统计学意义(P<0.05),与空白对照组(0.34±0.06)差异无统计学意义(P>0.05)。RES+碘酸钠损伤组Bcl-2蛋白的相对表达量(0.56±0.14)与碘酸钠损伤组(0.24±0.04)差异有统计学意义(P<0.05),与空白对照组(0.73±0.08)差异无统计学意义(P>0.05)。Hosste33342检测结果显示,RES+碘酸钠损伤组凋亡细胞含量明显低于碘酸钠损伤组(P<0.05),且流式细胞术检测发现,RES+碘酸钠损伤组和碘酸钠损伤组比较,RES+碘酸钠损伤组早期凋亡的细胞含量相对较高,晚期凋亡细胞含量相对较少。结论RES对碘酸钠诱导损伤的RPE细胞的凋亡具有良好的抑制作用,对干性AMD的治疗具有潜在价值。

Objective To study the inhibitory effect of resveratrol(RES) on sodium iodate-induced apoptosis of human retinal pigment epithelial(RPE) cells and its mechanism,and to explore whether RES can be used in the treatment of dry age-related macular degeneration(AMD).Methods RPE cells induced by sodium iodate were used to mimic dry AMD.RPE cells were divided into three groups:blank control group,sodium iodate injury group and RES+sodium iodate injury group.Cell morphology was observed in each group.MTT assay was used to detect the optimal concentration of RES on RPE cells and the protective effect of RES on RPE cells damaged by different concentrations of sodium iodate,and Western blot was used to detect the expression of apoptotic proteins Bax and Bcl-2.Hoechst33342 and flow cytometry were used to detect the changes in total apoptotic cells and the differences of apoptotic types.Results RES had a strong protective effect on RPE cells at the concentration of 5 μmol·L^-1,and had a strong protective effect on RPE cells with different degrees of injury.Relative expression of Bax protein in RES+sodium iodate injury group(0.45±0.07) was significantly lower than that in sodium iodate injury group(0.90±0.09) and the difference was statistically significant(P<0.05).There was no significant difference between sodium iodate injury group and blank control group(0.34±0.06)(P>0.05).Relative expression of Bcl-2 protein in RES+sodium iodate injury group(0.56±0.14) was significantly higher than that in sodium iodate injury group(0.24±0.04).There was a significant increase(P<0.05),but there was no significant difference compared with the blank control group(0.73±0.08).The results of Hoechst33342 showed that the content of apoptotic cells in the RES+sodium iodate injury group was significantly lower than that in sodium iodate injury group.Flow cytometry showed that the content of apoptotic cells in RES+sodium iodate injury group was relatively higher in the early stage than that in sodium iodate injury group,while the content of apoptotic cells in the late stage was relatively less.Conclusion RES can inhibit the apoptosis of RPE cells induced by sodium iodate and have potential value in the treatment of dry AMD.