中文摘要：

【目的】探究过氧化氢（H2O2）对骨髓间充质干细胞（BMSCs）氧化应激损伤所致衰老的相关机制,摸索合适的H2O2衰老造模浓度,为后期的药物干预实验研究奠定基础。【方法】采用全骨髓贴壁法培养BMSCs,H2O2分别以100、200、300、400、500、600、700、800、1 000μmol/L浓度作用于BMSCs 2、12、24、48 h,采用四甲基偶氮唑盐（MTT）法检测细胞代谢活力;β-半乳糖苷酶染色判断细胞衰老的建立成功与否;采用反转录荧光定量技术（RT-q real-time PCR）观察衰老相关基因p16、p21、p53表达水平的变化。【结果】当H2O2浓度〉700μmol/L时,BMSCs凋亡明显升高,而H2O2浓度〈400μmol/L时,变化不明显,甚至有刺激BMSCs增殖的作用;以500μmol/L H2O2作用BMSCs 24 h后,β-半乳糖苷酶染色阳性率显著增加,p21、p16、p53基因表达显著升高,差异均具有统计学意义（P〈0.05或P〈0.01）。【结论】以500μmol/L H2O2浓度作用于BMSCs 24 h后,可造成BMSCs氧化损伤衰老模型,并可使衰老相关基因p16、p21、p53表达上调。

英文摘要：

Objective To study the aging mechanism of bone marrow mesenchymal stem ceils （BMSCs） due to the oxidative stress damage induced by hydrogen peroxide （H2O2） , and to explore the appropriate H2O2 concentration for establishing aging model, which will lay the foundation for the later research of intervention drug. Methods BMSCs were obtained by whole bone marrow adherent culture method, and then were cultured with H2O2 at the concentrations of 100, 200, 300, 400, 500, 600, 700, 800, 1 000 μmol/L for 2, 12, 24, and 48 h, respectively. The cell metabolic activity was detected with methyl thiazolyl tetrazolium （MTT） assay, β- galactosidase staining was applied to confirm the cell senescence, and fluorescent quantitative reverse transcription technology （RT-q real-time PCR） was adopted to observe the expression levels of aging-related gene p16, p21, and p53 . Results When the H2O2 concentration was over 700 μmol/L, BMSCs apoptosis was significantly increased. However, H2O2 at the concentration of less than 400 μmol/L had no effect on BMSCs apoptosis, and even stimulated their growth. After cultured with H202 at the concentration of 500 μmol/L for 24 h, BMSCs had higher βgalactosidase staining positive rate, and had higher expression levels of p21, p16, p53 （P〈0.05 or P〈0.01 compared with those in the blank control group） . Conclusion Culturing with H202 at the concentration of 500 μmol/L for 24 h can induce the oxidative stress damage aging model, and can up-regulate the expression of aging-related gene p16, p21, and p53.