姜黄素对高浓度葡萄糖培养的RF/6A细胞活力的影响

Effects of curcumin on viability of high-glucose- concentration-cultured RF/6A

摘要:

目的:探讨姜黄素对高浓度葡萄糖培养的猴脉络膜-视网膜内皮细胞（rhesus choroidoretinal endothelial cell,RF/6A）活力的影响及其机制。方法:RF/6A分4组培养（n=6）:对照组；高浓度葡萄糖组（培养基添加30mmol/L葡萄糖）；姜黄素组（培养基添加30μmol/L姜黄素）；姜黄素-葡萄糖组（培养基添加30mmol/L葡萄糖及30μmol/L姜黄素）。以噻唑蓝比色法（methyl thiazolyl tetrazolium,MTT）检测培养1,3,5d后各组细胞活力的变化；试剂盒检测分组5d后各组细胞丙二醛（malonaldehyde,MDA）含量及超氧化物歧化酶（superoxide dismutase,SOD）活性；Western-blot检测分组5d后各组增殖细胞核抗原（proliferating cell nuclear antigen,PCNA）及抑癌基因p27蛋白（protein 27,p27）表达。结果:与对照组相比,高浓度葡萄糖组RF/6A活性明显降低,MDA含量增加,SOD活力降低,PCNA表达下调,p27表达上调（P＜0.01）；而姜黄素组及姜黄素-葡萄糖组RF/6A活力、MDA含量、SOD活力、PCNA与p27的表达均无明显变化（P＞0.05）。结论:姜黄素维持高浓度葡萄糖培养的RF/6A活力,可能与姜黄素抑制高浓度葡萄糖诱发的氧化应激,恢复RF/6A的PCNA及p27蛋白表达有关。

Abstract:

AIM:To explore effects of curcumin on viability of high glucose concentration (HGC) cultured rhesus choroido-retinal endothelial cell (RF/6A).METHODS:RF/6A was assigned into 4 groups (n= 6/group). Control group (CONT group), high glucose concentration group (HGC group):30mmol/L glucose was administered, curcumin group (CUR group):30μmol/L curcumin was administered, curcumin-glucose group (C-G group):30mmol/L glucose and 30μmol/L curcumin were administered. Cell viability was detected by methyl thiazolyl tetrazolium (MTT) at 1, 3 and 5 day after cultivation. Moreover, malonaldehyde (MDA), activity of superoxide dismutase (SOD) and expression of proliferating cell nuclear antigen (PCNA) and anti-oncogene protein 27 (p27) were detected at day 5.RESULTS:Compared with CONT group, HGC group showed decrease of cell viability, increase of MDA, decrease of activity of SOD, down-regulation of PCNA and up-regulation of p27 (P＜0.01). While, no difference could be detected in CUR group or C-G group compared with CONT group (P＞0.05). CONCLUSION:Curcumin inhabits HGC-induced oxidative stress, reverses expression of PCNA and p27, and, therefore, contribute to curcumin maintaining RF/6A viability.