背景:氧化应激诱导的视网膜色素上皮(RPE)细胞损伤是年龄相关性黄斑变性(AMD)的主要因素。维生素D3(VD3)是一种强大的抗氧化剂,已被认为具有抗衰老特性和治疗AMD的潜力。本研究旨在探讨VD3对RPE细胞氧化凋亡的影响,为AMD的治疗提供实验依据。
方法:人视网膜色素上皮细胞19(ARPE-19)细胞分为4组:空白组(未处理),模型组(在400μmol/LH2O2培养基中孵育1h),VD3组(在100μmol/LVD3培养基中孵育24h),和处理组(在400μmol/LH2O2和100μmol/LVD3的培养基中孵育1h)。细胞活力,细胞衰老,ROS含量,维生素D特异性受体的表达水平,Akt,Sirt1,NAMPT,和JNKmRNA表达水平,SOD活性,MDA,GSH,并测量GPX水平。
结果:我们首先建立了具有H2O2的ARPE-19细胞应激模型。我们的对照实验表明,VD3处理在6-48h内对ARPE-19细胞活力没有显着影响。用VD3处理应激的ARPE-19细胞显示出混合的结果;caspase-3表达降低,Bcl-2表达增加,ARPE-19细胞的MDA水平降低,GSH-PX,GPX和SOD水平升高,Akt的相对mRNA表达水平,Sirt1、NAMPT均升高(P<0.05),JNK的相对mRNA表达水平降低(P<0.05)。
结论:VD3可能会减缓AMD的发展。
BACKGROUND: Oxidative stress-induced retinal pigment epithelium (RPE) cell damage is a major factor in age-related macular degeneration (AMD). Vitamin D3 (VD3) is a powerful antioxidant and it has been suggested to have anti-aging properties and potential for treating AMD. This study aimed to investigate the effect of VD3 on RPE cell oxidative apoptosis of RPE cells in order to provide experimental evidence for the treatment of AMD.
METHODS: Human retinal pigment epithelial cell 19 (ARPE-19) cells were divided into four groups: blank group (untreated), model group (incubated in medium with 400 μmol/L H2O2 for 1 h), VD3 group (incubated in medium with 100 μmol/L VD3 for 24 h), and treatment group (incubated in medium with 400 μmol/L H2O2 for 1 h and 100 μmol/L VD3 for 24 h). Cell viability, cell senescence, ROS content, expression levels of vitamin D specific receptors, Akt, Sirt1, NAMPT, and JNK mRNA expression levels, SOD activity, and MDA, GSH, and GPX levels were measured.
RESULTS: We first established an ARPE-19 cell stress model with H2O2. Our control experiment showed that VD3 treatment had no significant effect on ARPE-19 cell viability within 6-48 h. Treating the stressed ARPE-19 cells with VD3 showed mixed results; caspase-3 expression was decreased, Bcl-2 expression was increased, MDA level of ARPE-19 cells was decreased, GSH-PX, GPX and SOD levels were increased, the relative mRNA expression levels of Akt, Sirt1, NAMPT were increased (P < 0.05), and the relative mRNA expression level of JNK was decreased (P < 0.05).
CONCLUSIONS: VD3 can potentially slow the development of AMD.