低浓度H2O2预处理增强骨髓间充质干细胞活力  

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

作　　者：张娅[1] 陈民佳[2] 朱明[2] 徐祥[2] 黄宏[2] 邱伟[2] 邢伟[2] 郭韡[2] 杨戎[1] 

机构地区：[1]重庆医科大学基础医学院干细胞与组织工程研究室,重庆400016 [2]第三军医大学第三附属医院野战外科研究所,创伤、烧伤与复合伤国家重点实验室,重庆400042

出　　处：《基础医学与临床》2016年第3期342-347,共6页Basic and Clinical Medicine

基　　金：国家重点基础研究发展规划项目(2012CB518105);国家自然科学基金(81372059,81571912)

摘　　要：目的探讨过氧化氢(H_2O_2)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H_2O_2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50μmol/L H_2O_2预处理BMSCs 12 h后,再经500μmol/L H_2O_2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/m TOR关键蛋白的表达。结果 25、50μmol/L H_2O_2能有效促进BMSCs增殖;50、100μmol/L H_2O_2能显著上调SDF-1、CXCR4、CXCR7表达;50μmol/L H_2O_2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;50μmol/L H_2O_2预处理干细胞可显著抑制500μmol/L H_2O_2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/m TOR通路关键蛋白磷酸化,CXCR7抗体则阻断此效应。结论H_2O_2预处理通过CXCR7受体活化Akt/m TOR信号通路增强干细胞存活和抗凋亡能力。Objective To investigate the effect and its mechanism of H202 preconditioning-induced the enhance- ment of anti-apoptosis in BMSCs. Methods BMSCs isolated from rat were treated with H202 for 48 h, or 50 μmol/L H202 proconditioning following treatment with 500 p, mol/L H202 combined with or without anti-CXCR7 antibody for 48 h. BMSCs viability was measured by MTF assay. The apoptosis was measured by Hoechst33342. The expression of SDF-1 and its CXCR4, CXCR7 receptor, Bcl-2, Bax and the proteins involved in Akt/mTOR pathway were detected by Western blot. Results 5 μmol/L and 50 μmol/L was promoted markedly H202 BMSCs proliferation and the expression of SDF- 1 and its CXCR4, CXCR7 receptor, and key phosphorylated proteins ofAkt/mTOR pathway. Transwell migration assay and scraping test showed that H202 preconditioning of BMSCs sig- nificantly augmented the migration ability, and protected BMSCs from 500 μmol/L H202-induced apoptosis, decrea- sing the rate of apoptotic cells, but antagonizing by anti-CXCR7 antibody. Conclusions H202 preconditioning en- hances the activity of bone marrow stem cell through CXCR7 receptor.

关 键 词：骨髓间充质干细胞 过氧化氢 预处理 CXCR7受体 细胞保护 

分 类 号：R457.7[医药卫生—治疗学]