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中国人民解放军总医院老年心血管病研究所
中国科技出版传媒股份有限公司
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中华老年多器官疾病杂志编辑委员会
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E-mail: zhlndqg@mode301.cn
创刊人 王士雯
总编辑 范利
副总编辑 陈韵岱
执行主编 叶大训
编辑部主任 王雪萍
ISSN 1671-5403
CN 11-4786
创刊时间 2002年
出版周期 月刊
邮发代号 82-408
友情链接
张正斌,朱丽雯,谭延振,冯攀,张冰,吴岩,易蔚,王晓明,孙阳.C1q/TNF相关蛋白3减轻氧化应激诱导的间充质干细胞衰老[J].中华老年多器官疾病杂志,2019,18(5):372~377
C1q/TNF相关蛋白3减轻氧化应激诱导的间充质干细胞衰老
C1q/TNF-related protein 3 attenuates oxidative stress-induced senescence of mesenchymal stem cells
投稿时间:2018-11-29  
DOI:10.11915/j.issn.1671-5403.2019.05.076
中文关键词:  C1q/TNF相关蛋白3;间充质干细胞;衰老;过氧化氢
英文关键词:C1q/TNF-related protein 3; mesenchymal stem cells; senescence; hydrogen peroxide This work was supported by the National Natural Science Foundation of China
基金项目:国家自然科学基金(81870266,81470477,81570231);陕西省创新人才推进计划-青年科技新星项目(2017KJXX-56)
作者单位E-mail
张正斌 空军军医大学第一附属医院老年病科,西安 71003  
朱丽雯 陕西中医药大学第二临床医学院,咸阳 712000  
谭延振 空军军医大学第一附属医院心血管外科,西安 710032  
冯攀 空军军医大学第一附属医院心血管外科,西安 710032  
张冰 空军军医大学第一附属医院心血管外科,西安 710032  
吴岩 空军军医大学第一附属医院心血管外科,西安 710032  
易蔚 空军军医大学第一附属医院心血管外科,西安 710032  
王晓明 空军军医大学第一附属医院老年病科,西安 71003  
孙阳 空军军医大学第一附属医院老年病科,西安 71003 dr_yangsun@163.com 
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中文摘要:
      目的 建立间充质干细胞(MSCs)衰老模型,探讨C1q/TNF相关蛋白3(CTRP3)对衰老MSCs的作用并初步探讨机制。方法 用过氧化氢(H2O2)诱导小鼠MSCs衰老,CCK8实验和β-半乳糖苷酶染色验证模型的建立。外源性给予CTRP3处理MSCs 24 h,CCK8实验检测增殖活性,β-半乳糖苷酶染色检测MSCs衰老,分化诱导实验检测MSCs分化能力,Western blotting法检测MSCs凋亡。RT-PCR检测MSCs中衰老相关基因P16、P21以及抗衰老基因SIRT1的mRNA表达变化。采用GraphPad Prism 6进行统计分析。组间比较采用方差分析或LSD两两比较。结果 H2O2可抑制MSCs的增殖活性,且呈浓度依赖性。与正常对照组相比,200 μmol/L H2O2作用4 h,细胞立体感消失,细胞形态不规则;CCK8染色结果显示细胞增殖活性显著下降;β-半乳糖苷酶阳性细胞百分比显著升高。继续CTRP3处理24 h后,衰老MSCs增殖和分化能力增强。Western blotting结果表明CTRP3可显著减低衰老MSCs的凋亡。RT-PCR检测发现CTRP3上调MSCs抗衰老基因SIRT1 mRNA表达,并且下调衰老相关基因P16、P21 mRNA的表达。结论 CTRP3可抑制MSCs衰老,SIRT1基因表达水平上调,及P16、P21基因表达水平下调可能是其重要机制。
英文摘要:
      Objective To explore the role of C1q/TNF-related protein 3 (CTRP3) in senescence model of mesenchymal stem cells (MSCs) and explore the mechanism. Methods The senescence model of mouse MSCs was induced by hydrogen peroxide (H2O2) treatment, and then identified by CCK8 assay and β-galactosidase staining. After exogenous treatment of CTRP3 was given to the obtained MSCs for 24 h, CCK8 was used to detect cell proliferation, and β-galactosidase staining was employed to measure the senescence. The differentiation capacity of the aging MSCs were measured by differentiation induction assay, and the expression of apoptosis-related proteins were detected by Western blotting. Then, RT-PCR was applied for the expression changes of the senescence-related genes P16, P21 and the anti-aging gene SIRT1. ANOVA or LSD analysis was used for intergroup comparison. Results H2O2 inhibited the proliferation activity of MSCs in a concentration-dependent manner. After 200 μmol/L H2O2 treatment for 4 h, light microscopy displayed that the 3-dimensional impression of the MSCs disappeared, and the cells presented irregular shape. CCK8 assay results showed that the cell proliferation activity was significantly decreased (P<0.05); while the percentage of β-galactosidase positive cells was obviously elevated. After 24 h of CTRP3 treatment, the proliferation and differentiation capacities of senescent MSCs were increased. Western blotting indicated that CTRP3 significantly reduced the apoptosis of senescent MSCs. RT-PCR analysis showed that CTRP3 up-regulated the MSCs anti-aging gene SIRT1 and down-regulated the expression of aging-related genes P16 and P21 at mRNA level. Conclusion CTRP3 treatment inhibits the apoptosis of senescent MSCs, which might be related with the up-regulation of SIRT1 and down-regulation of P16 and P21.
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