| 白藜芦醇通过保护线粒体功能延缓骨髓间充质干细胞衰老 |
| |
| 引用本文： | 张大勇,林九洲,王雅艳,许珊,罗成专,蔡佳烨,蒋学范,潘建平. 白藜芦醇通过保护线粒体功能延缓骨髓间充质干细胞衰老[J]. 浙江大学学报(医学版), 2019, 48(6): 617-624. DOI: 10.3785/j.issn.1008-9292.2019.12.05 |
| |
| 作者姓名： | 张大勇 林九洲 王雅艳 许珊 罗成专 蔡佳烨 蒋学范 潘建平 |
| |
| 作者单位： | 1. 浙江大学城市学院医学院, 浙江 杭州 3100152. 浙江省人民医院耳鼻喉科, 浙江 杭州 310014 |
| |
| 基金项目： | 浙江省自然科学基金(Y17H250001);国家自然科学基金(81671613);浙江省医药卫生科研计划(2016KYB024);浙江省中医药科学研究基金(2016ZA024);浙江省公益技术应用研究（实验动物）计划项目(LGD19C070001);杭州市农业与社会发展科研项目(20180533B30) |
| |
| 摘 要： | 目的: 探讨白藜芦醇对骨髓间充质干细胞（MSC）衰老的影响及其机制。方法: 分离SD乳鼠（7日龄）MSC，传代培养至第3代后检测0、1、10、50 g/L D-半乳糖对MSC衰老的影响，确定D-半乳糖诱导MSC衰老的最佳浓度。再将细胞随机分为10、50、100 μmol/L白藜芦醇组及对照组。衰老相关β-半乳糖苷酶（SA-β-gal）染色观察各组细胞衰老变化；DCFH-DA染色检测总活性氧水平；MitoSOX Red染色检测线粒体活性氧水平；线粒体膜电位检测试剂盒（JC-1法）检测线粒体膜电位变化；纯化线粒体膜通道孔（mPTP）比色法检测线粒体膜通道开放水平；蛋白质印迹法检测衰老相关蛋白p53、p16、γ-H2AX表达和细胞质内线粒体细胞色素C外泄水平。结果: 10、50 g/L D-半乳糖可明显增加SA-β-gal染色阳性MSC数量和线粒体活性氧水平（均P < 0.01）。与对照组比较，白藜芦醇组SA-β-gal染色阳性细胞数减少（均P < 0.01），p53、p16和γ-H2AX表达减少，细胞内总活性氧和线粒体活性氧水平下降（均P < 0.01），线粒体膜电位下降（P < 0.05或P < 0.01），膜通道开放水平降低（P < 0.05或P < 0.01），细胞质内细胞色素C外泄减少。结论: 白藜芦醇可保护MSC线粒体功能，延缓MSC衰老。
|
| 关 键 词： | 白藜芦醇 骨髓细胞 间质干细胞 线粒体 衰老 细胞 培养的 |
| 收稿时间： | 2019-01-30 |
Effects of resveratrol on aging of mesenchymal stem cells and its mechanism |
| |
| ZHANG Dayong,LIN Jiuzhou,WANG Yayan,XU Shan,LUO Chengzhuan,CAI Jiaye,JIANG Xuefan,PAN Jianping. Effects of resveratrol on aging of mesenchymal stem cells and its mechanism[J]. Journal of Zhejiang University. Medical sciences, 2019, 48(6): 617-624. DOI: 10.3785/j.issn.1008-9292.2019.12.05 |
| |
| Authors: | ZHANG Dayong LIN Jiuzhou WANG Yayan XU Shan LUO Chengzhuan CAI Jiaye JIANG Xuefan PAN Jianping |
| |
| Affiliation: | 1. School of Medicine, Zhejiang University City College, Hangzhou 310015, China2. Department of Otolaryngology, Zhejiang Provincial People's Hospital, Hangzhou 310014, China |
| |
| Abstract: | Objective: To investigate the effects of resveratrol (Res) on aging of marrow mesenchymal stem cells (MSCs), and to explore its mechanism. Methods: MSCs were isolated from young SD rats and cultured in vitro. The optimal D-gal concentration for induction of MSCs senescence was determined. Then MSCs were randomly divided into four groups, namely the control group, 10μmol/L, 50μmol/L and 100μmol/L Res groups. After the cells were treated with different concentration of Res for 48 h, the senescence-associated changes were examined with senescence-associated-β-galactosidase (SA-β-gal) staining; the expression of p53, p16 and γ-H2AX was evaluated by Western blot. The total active oxygen species (ROS) level was determined by flow cytometry with DCFH-DA staining. In order to assess the effect of Res on the mitochondrial function, MitoSox Red staining was used to detect mitochondrial ROS levels in each group, mitochondrial membrane potential was detected by JC-1 assay, mPTP method was used to detect mitochondrial membrane channel opening level, and Western blot was used to detect the expression level of cytoplasmic cytochrome C (Cyt-C). Results: D-gal 10 and 50 g/L significantly increased the number of SA-β-gal positive cells and the level of mitochondrial ROS (all P < 0.01). Therefore, 10 g/L D-gal was used to induce the senescence of MSCs in subsequent experiment. Compared with the control group, the number of SA-β-gal positive cells in Res groups significantly decreased (all P < 0.01), the expression of p53, p16 and γ-H2AX decreased, and the total and mitochondrial ROS level also decreased (all P < 0.01). Moreover, mitochondrial membrane potential, open level of mitochondrial membrane channels and the levels of cytoplasm Cyt-C in the Res treatment groups decreased compared with the control group (P < 0.05 or P < 0.01). Conclusion: Resveratrol can protect the mitochondrial function of MSCs, and effectively delay the MSC senescence. |
| |
| Keywords: | Resveratrol Bone marrow cells Mesenchymal stem cells Mitochondria Aging Cells cultured |
|
| 点击此处可从《浙江大学学报(医学版)》浏览原始摘要信息 |
|
点击此处可从《浙江大学学报(医学版)》下载全文 |
|
