缺氧骨髓干细胞对糖尿病心肌病的保护作用  

The protective effect of bone marrow mesenchymal stem cells by anoxic preconditioning on diabeticcardiomyopathy

作　　者：李佳慧[1] 张楠[2] 乔薇[3] 王勇[1] 王建安[4] 

机构地区：[1]中日友好医院心内科,北京100029 [2]浙江大学医学院附属邵逸夫医院内分泌科 [3]中日友好医院干部保健科 [4]浙江大学医学院附属第二医院心内科

出　　处：《中华急诊医学杂志》2011年第10期1062-1066,共5页Chinese Journal of Emergency Medicine

摘　　要：目的探讨缺氧预处理骨髓间充质干细胞（AP-MSC）对大鼠糖尿病心肌病（DCM）的保护作用。方法8周雄性SD大鼠腹腔单次注射链脲佐菌素（STZ）（60mg／kg）构建糖尿病（DM）模型。骨髓间充质干细胞（MSC）由密度梯度离心法分离获取，并经3～5次传代扩增和纯化。MSC在体外用CM—DiI（2μg／mL）标记20min。AP-MSC缺氧（氧体积分数〈0．5％）3h。STZ注射后4个月，大鼠随机（随机数字法）分组，分别心肌注射150μL DMEM，5×10^6 MSC／150μL，或5×10^6AP-MSC／150μL（每组n=10）。在STZ注射后3个月及MSC移植后2周，采用心脏超声评价大鼠心功能情况，并在移植后2周，采用碱性磷酸酶染色，以凋亡相关蛋白和信号通路蛋白的Westernblot分析来评价三组大鼠的心脏情况。结果与DMEM相比，MSC特别是AP-MSC增加了DCM大鼠的短轴缩短率（P〈0．01），AP-MSC显著增加了DCM的心肌毛细血管密度（P）。AP-MSC组Bcl-2／Bax比值升高（P〈0．05vs．DMEM），被剪切的caspase-3表达低于DMEM和MSC组（P〈0．01）。结论AP-MSC对大鼠DCM具有保护作用，改善了心功能。Objective To explore the protective effect of bone marrow mesenchymal stem cells（MSC） on diabetic myocardium and anoxic pre-conditioning （AP）. Methods Eight-week-- old male Sprague-Dawley rats were given with a single intraperitoneal injection of streptozotocin （STZ） （60 mg/kg） to induce diabetes mellitus （DM）. Donor rats were 8-week-old male Sprague - Dawley rats. Before transplantation, MSC were incubated in CM-DiI at a concentration of 2 μg/mL for 20 min. AP-MSC were exposed to 3 hours of anoxia. At 4 months after STZ injection, diabetic rats were randomly given with an intramyocardial injection of one of the followings ： 150 μL of Dulbecco＇s modified Eagleg medium （DMEM） , 5 × 10^6 MSC/150 μL, or 5 × 10^6 AP - MSC/150 μL （ n = 10 for each group）. Three months after STZ injection and 2 weeks after transplantation, we evaluated the cardiac function by echocardiography, and also evaluated the cardiac conditions by alkaline phosphatase staining, western blot analysis for apoptosis related proteins and signal pathways. Results MSC, especially AP - MSC increased fractional shortening （FS） of diabetic heart （P 〈 0. 01 vs DMEM respectively）. AP-MSC greatly increased the capillary density of diabetic myocardium （P 〈0. 01 vs DMEM and MSC group respectively）. AP-MSC are anti-apoptotic in the rat DCM model, possibly mediated through cardiac upregulation of Bcl-2/Bax ratio （ P 〈 0. 05 ） and inhibiting the expression and activation of caspase - 3 （ P 〈 0. 01 ）. Conclusions Intramyocardial transplantation of AP - MSC has a protective effect on diabetic cardiomyopathy.

关 键 词：骨髓间充质干细胞 糖尿病心肌病 糖尿病 缺氧 缺氧预处理 

分 类 号：R587.2[医药卫生—内分泌]