低浓度H2O2预处理增强骨髓间充质干细胞活力

基础医学与临床 ›› 2016, Vol. 36 ›› Issue (3): 342-347.

低浓度H2O2预处理增强骨髓间充质干细胞活力

张娅¹,陈民佳²,朱明²,徐祥³,黄宏³,邱伟²,邢伟³,郭韡²,杨戎¹

1. 重庆医科大学
2. 第三军医大学第三附属医院
3. 第三军医大学大坪医院

收稿日期:2015-09-24 修回日期:2015-12-07 出版日期:2016-03-05 发布日期:2016-02-22
通讯作者: 杨戎 E-mail:1364262572@qq.com
基金资助:
国家自然基金;国家自然基金

The proliferation of BMSCs is increased following the precondition with low concentration hydrogen peroxide

Received:2015-09-24 Revised:2015-12-07 Online:2016-03-05 Published:2016-02-22
Supported by:
The National Nature Science Foundation of China;The National Nature Science Foundation of China

摘要/Abstract

摘要： 目的探讨过氧化氢（H2O2）预处理增强骨髓间充质干细胞（BMSCs）存活和抗凋亡能力的作用及机制。方法分离、培养、鉴定小鼠BMSCs。观察不同低浓度H2O2处理BMSCs 48 h后细胞的增殖，以及SDF-1及其受体CXCR4、CXCR7的表达；其次，观察经50 μmol/L H2O2预处理BMSCs12 h后，再经500 μmol/L H2O2作用，或同时加入CXCR7抗体作用48 h，用Hoechst33342染核观察BMSCs凋亡率，Western blot检测Bcl-2、Bax、CXCR4、CXCR7和信号通路Akt/mTOR关键蛋白的表达。结果 25、50 μmol/L H2O2能有效促进BMSCs增殖；50、100 μmol/L H2O2能显著上调SDF-1、CXCR4、CXCR7表达；50 μmol/L H2O2预处理干细胞能显著增强迁移能力，而CXCR7抗体能阻断此效应；50 μmol/L H2O2预处理干细胞可显著抑制500 μmol/L H2O2引起的损伤，凋亡细胞核由30.97％±3.71％降至16.23％±2.51％（p<0.01），但仍然显著高于对照组（4.67％±2.37％）（p<0.01）；同样，预处理能显著下调Bax、上调Bcl-2表达（p<0.05），上调Akt/mTOR通路关键蛋白磷酸化，CXCR7抗体则阻断此效应。结论 H2O2预处理通过CXCR7受体活化Akt/mTOR信号通路增强干细胞存活和抗凋亡能力。

关键词: 骨髓间质干细胞, 过氧化氢, 预处理, CXCR7受体, 细胞保护

Abstract: Objective To investigate the effect and its mechanism of H2O2 preconditioning-induced the enhancement of anti-apoptosis in BMSCs. Methods BMSCs isolated from rat were treated with H2O2 for 48h, or 50μmol/L H2O2 proconditioning following treatment with 500μmol/L H2O2 combined with or without anti-CXCR7 antibody for 48h. BMSCs viability was measured by MTT assay. The apoptotic cells was measured by Hoechst33342. The expression of SDF-1 and its CXCR4, CXCR7 receptor, Bcl-2, Bax and the key proteins of Akt/mTOR pathway was detected by Western blot. Results 25μmol/L and 50μmol/L was promoted markedly H2O2 BMSCs proliferation and the expression of SDF-1 and its CXCR4, CXCR7 receptor, and key phosphorylated proteins of Akt/mTOR pathway. Transwell migration assay and scraping result were shown that H2O2 preconditioning of BMSCs significantly augmented the migration ability, and protected BMSCs from 500 μmol /L H2O2-induced apoptosis，decreasing the rate of apoptotic cells, but antagonizing by anti-CXCR7 antibody. Conclusion H2O2 preconditioning induces the enhancement of the activity of bone marrow stem cell through CXCR7 receptor.

Key words: Bone marrow derived stromal stem cells(BMSCs), Hydrogen peroxide (H2O2), precondition, CXCR7 receptor, Cytoprotection

张娅陈民佳朱明徐祥黄宏邱伟邢伟郭韡杨戎. 低浓度H2O2预处理增强骨髓间充质干细胞活力[J]. 基础医学与临床, 2016, 36(3): 342-347.

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