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| IGF-1对2,5-己二酮诱导的骨髓间充质干细胞凋亡的拮抗作用研究 |
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白秋芳1, 邹明1, 张继志2, 刘卓3, 朴丰源4
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1.大连大学附属中山医院 药剂科, 辽宁 大连 116001;2.大连大学附属中山医院 神经外科, 辽宁 大连 116001;3.锦州医科大学 基础医学实验中心机能学实验平台, 辽宁 锦州 121001;4.大连大学附属中山医院 中心实验室, 辽宁 大连 116001
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| 摘要: |
| 目的 研究胰岛素样生长因子-1（IGF-1）对2，5-己二酮（HD）暴露所致的骨髓间充质干细胞（BMSCs）凋亡的拮抗作用及其机制。方法 将BMSCs随机分成对照组（Control组）、IGF组、染毒组（HD组）和3个干预组（IGF+HD组），即用生理盐水、50 ng/L IGF-1、40 mmol/L HD及不同浓度IGF-1（50，75，100 ng/L）共同处理24 h。然后，通过TUNEL法检测BMSCs的凋亡，应用试剂盒测定caspase-3的活性，应用western blot技术检测Akt和Bad的蛋白表达及磷酸化水平。结果 HD组BMSCs凋亡率及caspase-3活性显著高于Control组（P<0.05）。而经IGF-1干预后，上述指标显著低于HD组（P<0.05）。同时，HD组BMSCs的Akt和Bad蛋白磷酸化水平显著低于Control组（P<0.05）。而经IGF-1干预后，Akt和Bad的蛋白磷酸化水平显著高于HD组（P<0.05）。此外，各组间的Akt和Bad总蛋白的表达水平无统计学差异（P>0.05）。结论 IGF-1可通过调控Akt/Bad信号通路的活性，拮抗HD诱导的BMSCs凋亡。 |
| 关键词: 胰岛素样生长因子-1 骨髓间充质干细胞 抗凋亡 Akt/Bad信号通路 |
| DOI：10.11724/jdmu.2020.01.02 |
| 分类号:R96 |
| 基金项目:国家自然科学基金项目（81273038；8773402）；辽宁省科技厅自然基金指导计划（2019-ZD-0818） |
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| Anti-apoptotic effect of IGF-1 on bone mesenchymal marrow stem cells exposed to 2,5-hexanedione |
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BAI Qiufang1, ZOU Ming1, ZHANG Jizhi2, LIU Zhuo3, PIAO Fengyuan4
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1.Department of Pharmacy, Affiliated Zhongshan Hospital of Dalian University, Dalian 116001, China;2.Department of Neurosurgery, Affiliated Zhongshan Hospital of Dalian University, Dalian 116001, China;3.Functional Experiment Center of Basic Medical College, Jinzhou Medical University, Jinzhou 121001, China;4.Comprehensive Laboratory, Affiliated Zhongshan Hospital of Dalian University, Dalian 116001, China
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| Abstract: |
| Objective To investigate the anti-apoptotic effect of (insulin-like growth factor-1,IGF-1) on bone marrow mesenchymal stem cells(BMSCs) exposed to 2, 5-hexanedione(HD) and its protective mechanism. Methods BMSCs were randomly divided into control group, IGF-1 group, exposure group (HD group) and 3 IGF-1 antagonist groups (IGF+HD group), that is, the cells were treated with saline, 50 ng/L IGF-1, 40 mmol/L HD alone or 40 mmol/L HD with different concentrations of IGF-1 (50, 75, 100 ng/L) for 24 h. Then the apoptosis of BMSCs was estimated by TUNEL assay, caspase-3 activity was determined by commercial kit and the expression and phosphorylation level of Akt and Bad were examined by western blot. Results The apoptotic index and caspase-3 activity of BMSCs in HD group were significantly higher than those in Control group (P<0.05). However, after treating with IGF-1, the apoptotic index and caspase-3 activity of BMSCs were significantly lower than those in HD group (P<0.05). Morever, the phosphorylation level of Akt and Bad in BMSCs of HD group was significantly lower than those in Control group (P<0.05), while the phosphorylation level of Akt and Bad were significantly higher than those in HD group after treating with IGF-1 (P<0.05). Additionally, there were no significant differences in the the expression of total Akt and Bad among the groups (P>0.05). Conclusion IGF-1 may antagonize HD-induced BMSCs apoptosis by regulating the Akt/Bad signaling pathway. |
| Key words: IGF-1 BMSCs 2,5-hexanedione anti-apoptosis Akt/Bad pathway |