p53抑制剂对扩增晚期人骨髓间充质干细胞活力的影响  

Effect of p53 inhibitor on viability of human bone marrow mesenchymal stem cells in late-phase amplification

作　　者：贺泽斌 赵云鹤[1] 杨桂姣[1] 陆利[1] 

机构地区：[1]山西医科大学人体解剖学教研室,山西省太原市030001

出　　处：《中国组织工程研究》2015年第23期3616-3620,共5页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81200254);山西省回国留学人员科研资助项目(2014-033)~~

摘　　要：背景:p53抑制剂能否直接干预骨髓间充质干细胞活力及其可能的机制尚不完全清楚。目的:探讨p53抑制剂PFT-α对体外扩增培养晚期骨髓间充质干细胞衰老进程的影响,寻找延缓人骨髓间充质干细胞复制性衰老的关键靶点。方法:qP CR检测扩增早、晚期人骨髓间充质干细胞p53、p21和p15 mR NA的表达。20μmol/L p53抑制剂PFT-α或等量二甲基亚砜分别作用晚期人骨髓间充质干细胞2周,β-半乳糖苷酶(SA-β-Gal)染色观察衰老细胞阳性率,TUNEL染色法检测细胞凋亡情况。300μmol/L H2O2作用于人骨髓间充质干细胞30 min,CCK-8法检测细胞抗氧化应激能力。结果与结论:qP CR显示晚期人骨髓间充质干细胞p15,p21,p53 mR NA表达水平分别较早期人骨髓间充质干细胞增高(1.45±0.23)倍,(1.51±0.14)倍,(1.78±0.14)倍(P<0.05)。PFT-α组衰老细胞阳性率(41±5)%低于二甲基亚砜组(63±7)%(P<0.05),但PFT-α组与二甲基亚砜组细胞凋亡率差异无显著性意义(P>0.05)。经H2O2处理后,CCK-8结果显示PFT-α组吸光度值为二甲基亚砜组(1.27±0.13)倍(P<0.001),以上结果表明p53信号通路激活可能是导致人骨髓间充质干细胞衰老的重要因素,应用p53抑制剂PFT-α能够增强晚期人骨髓间充质干细胞抗氧化应激损伤能力。BACKGROUND： It is not fully understood that whether p53 inhibitor can directly intervene in the viability of bone marrow mesenchymal stem cells and the possible mechanism.OBJECTIVE： To investigate the effect of p53 inhibitor, PFT-α, on the aging process of bone marrow mesenchymal stem cells in late-phase amplification and to discover the key target to delay the replicative senescence of human bone marrow mesenchymal stem cells.METHODS： The expression levels of p53, p21, and p15 m RNA in human bone marrow mesenchymal stem cells in both early and late-phase amplification were detected by quantitative PCR assay. Then, human bone marrow mesenchymal stem cells in late-phase amplification were respectively treated with 20 μmol/L PFT-α or an equivalent amount of dimethyl sulfoxide for 2 weeks. The positive rate of aging cells was determined by SA-β-Gal staining. The apoptosis was detected by TUNEL staining. Human bone marrow mesenchymal stem cells were treated with 300 μmol/L H2O2 for 30 minutes, and then cellular anti-oxidative stress capacity was detected by cell counting kit-8 assay.RESULTS AND CONCLUSION： The quantitative PCR assay showed that the mR NA expression level of p15,p21 and p53 in human bone marrow mesenchymal stem cells in late-phase amplification was significantly increased（1.45±0.23）,（1.51±0.14） and（1.78±0.14） times as much as that in early phase amplification（P〈0.05）. The positive rate of aging cells in PFT-α group was significantly lower than that in the dimethyl sulfoxide group [（41±5）% vs.（63±7）%, P〈0.05）]. However, there was no significant difference in apoptosis rate between PFT-α group and dimethyl sulfoxide group.After treatment with H2O2, the absorbance value in the PFT-α group was（1.27±0.13） times as much as that in the dimethyl sulfoxide group（P〈0.001）. The above results demonstrate that the activation of p53 signaling pathway may be an important factor of causing aging of human bone marrow mesenchymal stem cells. Application of p

关 键 词：骨髓 间质干细胞 细胞凋亡 细胞衰老 基因 p53 干细胞 骨髓干细胞 复制性衰老 骨髓间充质干细胞 p53 PFT-Α 国家自然科学基金 

分 类 号：R394.2[医药卫生—医学遗传学]