紫绀及非紫绀型先心病患者来源的人骨髓间充质干细胞抗凋亡能力的比较  

Comparison of Anti-apoptotic Potency between Acyanotic and Cyanotic Congenital Heart Disease-derived Human Mesenchymal Stem Cells in vitro

作　　者：郭海娇 康凯[1] 刘焕欣[1] 刘思雨[1] 啜俊波[1] 蔡俊[1] 吴华[1] 李建中[1] 蒋树林[1] 王宜淳 

机构地区：[1]哈尔滨医科大学附属二院心脏外科心肌缺血机理与诊疗技术省部共建教育部重点实验室,哈尔滨150086 [2]大连枫叶国际学校,辽宁大连116000

出　　处：《中国胸心血管外科临床杂志》2016年第12期1172-1176,共5页Chinese Journal of Clinical Thoracic and Cardiovascular Surgery

基　　金：国家自然科学基金(81471805);哈尔滨市科技局创新人才科研基金(RC2016QN004036);黑龙江省大学生创新训练计划项目(201510226057)~~

摘　　要：目的 对比紫绀型先心病（cyanotic congenital heart disease,C-CHD）和非紫绀型先心病（acyanotic congenital heart disease,A-CHD）患儿来源的骨髓间充质干细胞（human mesenchymal stem cells,hMSCs）在乏氧环境下的抗凋亡能力并探讨其发生机制。方法 hMSCs取自C-CHD（C组）和A-CHD（A组）患儿（男女不限,0~5岁）,于体外乏氧环境下（1%O_2,5%CO_2,94%N_2）培养。以Annexin V/PI双染结合流式细胞技术对比两组细胞抵御缺血缺氧诱导细胞凋亡的能力;以Western blot法检测两组细胞中B细胞淋巴瘤-2基因（B-cell lymphoma-2,Bcl-2）,Bcl-2相关X蛋白（Bax）以及半胱氨酸天冬氨酸蛋白酶-3（caspase-3）的含量。结果 Annexin V/PI双染结合流式细胞检测结果显示：在缺血缺氧环境下,C组细胞的早期凋亡率（P〈0.01）和总体凋亡率（P〈0.05）低于A组细胞。Western-blot检测结果显示：C组细胞的凋亡抑制基因Bcl-2的含量高于A组细胞（P〈0.05）,抗凋亡抑制基因Bax（P〈0.05）以及caspase-3（P〈0.05）的含量低于A组细胞。结论 C-CHD患者来源的hMSCs具有更好的抗缺血缺氧诱导的总凋亡及早期凋亡能力,这可能与其天然乏氧诱导的Bcl-2表达上调,Bax,caspase-3表达下调有关。Objective To compare the anti-apoptotic potency of human mesenchymal stem cells （hMSCs） derived from patients with cyanotic congenital heart diseases （C-CHD） or acyanotic congenital heart diseases （A-CHD） in vitro and explore the possible mechanism. Methods hMSCs were isolated from patients with cyanotic （Group C） or acyanotic （Group A） congenital heart diseases and cultured in a hypoxic incubator （1% 02, 5% CO2, 94% N2） in vitro. The anti-apoptotic potency of the hMSCs was assayed by the Annexin V-FITC/PI double labeled flow cytometry. The content of B-cell lymphoma-2 （Bcl-2）, Bax and caspase-3 in both groups was determined by Western blot. Results Flow cytometry results revealed that hMSCs from C-CHD patients presented higher level of resistance to ischemia- and anoxia- induced apoptosis with lower overall （P〈0.05） and early apoptosis ratio （P〈0.01）. Further Western blot examination identified that C-CHD-derived hMSCs produced more Bcl-2 （P〈0.05） but less Bax （P〈0.05） and caspase-3 （P〈0.05） in comparison to their A-CHD-derived ones. Conclusion C-CHD-derived hMSCs presented the superiority for the antiapoptotic potential, and the possible mechanism is the favorable change of Bcl-2, Bax and caspase-3 induced by the natural hypoxic and anoxic precondition.

关 键 词：紫绀 骨髓干细胞 先天性心脏病 细胞凋亡 

分 类 号：R725.4[医药卫生—儿科]