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

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。

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 （P0.01）. While, no difference could be detected in CUR group or C-G group compared with CONT group （P0.05）.CONCLUSION：Curcumin inhabits HGC-induced oxidative stress, reverses expression of PCNA and p27, and, therefore, contribute to curcumin maintaining RF/6A viability.