| D-半乳糖诱导间充质干细胞衰老的研究 |
| 投稿时间：2018-01-26 修订日期：2018-03-16 点此下载全文 |
| 引用本文：杨孟恺,易诚青.D-半乳糖诱导间充质干细胞衰老的研究[J].医学研究杂志,2018,47(7):31-36 |
| DOI：
10.11969/j.issn.1673-548X.2018.07.008 |
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| 基金项目:国家自然科学基金资助项目（81371979） |
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| 中文摘要:目的 利用D-半乳糖诱导建立间充质干细胞（mesenchymal stem cells，MSCs）体外衰老模型，并探讨其潜在机制。方法 以正常培养的MSCs做对照组，以分别用1、10、100g/L D-半乳糖诱导的MSCs作为处理组，培养48h后，采用免疫蛋白印迹法（Western blot）检测细胞内AKT与mTOR蛋白磷酸化水平以及衰老相关蛋白p16Ink4a的变化。同时使用β-半乳糖苷酶试剂盒检测MSCs中β-半乳糖苷酶（SA-β-gal）表达量的变化。最后用AKT抑制剂预处理D-半乳糖诱导组以明确AKT/mTOR信号通路在MSCs衰老中的作用。结果 与对照组相比，10g/L D-半乳糖作用48h后MSCs中β-半乳糖苷酶表达和衰老相关蛋白p16Ink4a含量明显增加，同时p-AKT与p-mTOR含量也较对照组增加，加入AKT抑制剂后，衰老组β-半乳糖苷酶表达和衰老相关蛋白p16Ink4a含量皆有所下降。结论 D-半乳糖能成功诱导构建MSCs体外衰老模型，AKT/mTOR信号通路参与了这一过程。 |
| 中文关键词:D-半乳糖 信号通路 间充质干细胞 衰老 |
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| Study on D-galactose-induced Senescence of Mesenchymal Stem Cells |
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| Abstract:Objective To establish the aging model of mesenchymal stem cells (MSCs) induced by D-galactose (D-gal) and investigate its underlying mechanism. Methods The MSCs were divided into control group and D-gal treatment group. Control group was normal cultured MSCs. D-gal treatment group was the control group treated with different concentrations of D-gal (1, 10, 100g/L). All of them were treated for 48h. The expression of p16Ink4a protein, p-AKT, and p-mTOR were detected by Western blot. At the same time, the SA-β-galactose (SA-β-gal) staining was used to examined the senescence-associated changes. At last, AKT inhibitor was added in the D-gal treatment group to clarify the role of AKT/mTOR signaling pathway in MSCs aging. Results Compared with the control group, the quantity of SA-β-gal positive cells and the expression of p16Ink4a in MSCs were significantly increased in 10g/L D-gal treatment group. Meanwhile the expression of p-AKT and p-mTOR also increased compared with the control group. After adding in AKT inhibitor, the quantity of SA-β-gal positive cells in D-gal treatment group and the expression of p16Ink4a were decreased, too. Conclusion D-gal can successfully establish the aging MSCs model in vitro and AKT/mTOR signaling pathway is involved in this process. |
| keywords:D-galactose Signaling pathway Mesenchymal stem cells Senescence |
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