摘要
目的研究鼠尾草酸(carnosic acid,CA)延缓细胞衰老的作用及相关机制。方法采用体外细胞模型人胚肺二倍体成纤维2BS细胞系,应用四甲基偶氮唑盐(MTT)法测定CA对2BS细胞活力的影响,筛选获得最佳CA培养浓度,再用该浓度的CA对2BS进行传代培养,观察其对细胞代龄、衰老相关β-半乳糖苷酶(SA-β-Gal)活性、脂质过氧化产物丙二醛(MDA)水平、细胞周期分布和衰老相关蛋白p53、p21的影响。结果 MTT实验表明CA在1μmol/L、5μmol/L能够上调2BS细胞活力;采用含1μmol/L CA的培养液对2BS细胞从30PD(population doubling,倍增次数,亦称作"代")开始培养,直至完全停止生长,同时设立对照。对照组细胞于54PD左右发生衰老,SA-β-Gal染色阳性率达85%,而55PD的CA处理组仅为20%左右,接近年轻细胞水平;此时CA组细胞内MDA水平显著低于对照组,处于G1/G0期细胞的比例显著低于对照组,而S期比例则显著高于对照组,p53、p21蛋白水平均显著低于对照组,CA组细胞继续培养至61PD左右才出现衰老。结论 CA在1μmol/L剂量下能延缓2BS细胞的复制性衰老,该作用可能与其抗氧化能力有关。
Objective To investigate the effect of carnosic acid（CA） on cellular aging as its related mechanisms. Methods Human embryonic lung diploid fibroblasts 2BS cell line was used. MTT assay was performed to evaluate the effect of CA on cellular viability. An optimal concentration of CA was determined for 2BS cells culture from 30PD（population doubling）. The effects of CA were evaluated by detecting the changes of cumulative population doubling（CPD）, activity of senescence-associated beta galactosidase（SA-β-Gal）, malondialdehyde（MDA） level, cell cycle distribution as well as senescence associated molecules p53 and p21. Results It was showed in MTT assay that CA could increase the cellular viability at 1-5 μmol/L, with 1 μmol/L as the optimal dose. The SA-β-Gal positive rate was 20% in 2BS cells grown in medium with 1 μmol/L of CA when they reached 55PD, which was significantly lower than that in the control cells at 54PD（85%）. Meanwhile, a dramatic decline of intracellular MDA level was also observed in CA treated cells in comparison to that of control cells. Besides, CA treated cells showed a lower percentage of G1/G0 and high percentage of S distribution and decreased p53 and p21 protein expression. The 2BS cells grown in medium with 1 μmol/L CA reached about 61 CPD. Conclusion CA can delay the senescence of 2BS cells and this effect may be related with its antioxidant capacity at an optimal dose.
出处
《营养学报》
CAS
CSCD
北大核心
2014年第3期273-277,共5页
Acta Nutrimenta Sinica
基金
国家自然科学基金(No.31201040)
浙江省医药卫生一般研究计划项目(No.2013KYA003)
卫生部科学研究基金项目(No.WKJ2011-2-014)
关键词
鼠尾草酸
细胞衰老
氧化应激
carnosic acid
cellular senescence
oxidative stress
