重复远程缺血后适应对急性缺血心肌中骨髓间充质干细胞移植的影响  

Effects of repeated remote ischemic postconditioning on bone marrow mesenchymal stem cells transplantation in acute ischemic myocardium

作　　者：蒋钦[1] 向波[1] 于涛[1] 黄克力[1] 

机构地区：[1]四川省人民医院心脏外科中心,成都610072

出　　处：《中华细胞与干细胞杂志（电子版）》2016年第2期86-91,共6页Chinese Journal of Cell and Stem Cell（Electronic Edition）

基　　金：四川省人民医院博士基金(30305030842)

摘　　要：目的探讨在大鼠急性心肌缺血模型中重复远程缺血后适应对骨髓间充质干细胞(MSC)经心肌注射移植存留率的影响。方法结扎成年雌性SD大鼠的左前降支诱导心肌缺血。雄性SD大鼠来源的骨髓MSC培养至第三代。大鼠缺血30 min后开放结扎的冠状动脉,并经缺血心肌边缘注射4×106个骨髓间充质细胞。60只心肌缺血大鼠随机平均分为再灌注对照(IR)组、远程缺血后适应(RIPo C)组、重复远程缺血后适应(r RIPo C)组和CXCR4抗体拮抗(CXCR4-Ab)组。远程缺血后适应在后肢上实行4个周期5 min缺血和再灌注。重复远程缺血后适应在后肢上每3 d实行4个周期5 min缺血和再灌注。CXCR4抗体拮抗(CXCR4-Ab)组在行r RIPo C前经腹腔注射CXCR4特异的抗体。1个月以后,进行心功能评价、免疫组化寻找标记的细胞、聚合酶联反应检测Y染色体拷贝数计算心肌中细胞存留率,采用方差分析和独立t检验进行统计学分析。结果重复远程缺血后适应的细胞移植组显著改善缺血心脏的左室短轴缩短分数[r RIPo C(25.90±4.33)﹪,RIPo C(20.60±3.50)﹪,IR(16.60±3.20)﹪,CXCR4-Ab(20.00±3.23)﹪,F=11.422,P=0.000]和左室收缩末期内径[r RIPo C(5.24±0.51)mm,RIPo C(5.77±0.44)mm,IR(6.15±0.33)mm,CXCR4-Ab(5.78±0.33)mm,F=8.159,P=0.000]。免疫组化发现重复远程缺血后适应组的心脏内细胞滞留率更高,PCR检测MSC滞留比例[r RIPo C(2.33±0.46)﹪,RIPo C(1.85±0.50)﹪,IR(1.42±0.27)﹪,F=8.189,P=0.000];特异CXCR4抗体阻断降低重复远程缺血后适应诱导的心脏内高细胞滞留率[r RIPo C(2.33±0.46)﹪,CXCR4-Ab(1.82±0.36)﹪,n=10,P=0.014]和心脏收缩功能改善[r RIPo C(25.90±4.33)﹪,CXCR4-Ab(20.00±3.23)﹪,P=0.003]。结论重复远程缺血后适应较单次远程缺血后适应提高经心肌注射移植的骨髓MSC在心脏中的移植存留率。CXCR4受体在重复远程缺血后适应后期诱导的心肌高移植存留率中发挥重要作用。Objective To investigate the effect of repeated remote ischemic postconditioning （rRIPoC） on the retention of mesenchymal stem cells in the acute rat myocardial ischemia model. Methods Myocardial ischemia of adult female SD rats was induced by ligation of the left anterior descending （LAD） coronary artery for 30 minutes. Male SD rat-derived mesenehymal stem cell （MSC） were harvested from bone marrow. Cells of the third passage were used. A total of 4 ×10^6 MSC were intramyocardially infused into 60 female MI rats which were randomly assigned into IR, RIPoC, rRIPoC, and CXCR4-Ab groups. RIPoC group was induced with 4 cycles of 5-minute occlusion and reperfusion of the limb. rRIPoC group was induced with 4 cycles of 5-minute occlusion and reperfusion of the limb every three days. For CXCR4-Ab group, anti-CXCR4 antibody was intraperitoneally administered before rRIPoC procedure. One month later, all groups underwent left ventricular function examination. MSC engraftment in the myocardium was evaluated by immunohistochemistry （IHC） examination and quantitative polymerase chain reaction （PCR） for SRY gene in Y chromosome. Statistical analysis was conducted with two-way ANOVA and t test. Results MSC significantly ameliorated left ventricular shortening fraction [rRIPoC （25.90 ± 4.33） %, RIPoC （20.60± 3.50） %, IR （16.60 ± 3.20） %, CXCR4-Ab （20.00± 3.23 ） %, F = 11.422, P = 0.000] and left ventricular end-systolic diameter ErRIPoC （5.24 ± 0.51） mm, RIPoC （5.77 ± 0.44）mm, IR （6.15 ± 0.33）mm, CXCR4-Ab （5.78 ± 0.33） ram, F = 8.159, P = 0.000]. IHC examination for labeled infused cells and PCR analysis for Y chromosome demonstrated that rRIPoC significantly increased MSCs retention in myocardiumErRIPoC （2.33 ± 0.46） %, RIPoC （1.85 ± 0.50） %, IR （1.42 ± 0.27） %, F = 8.189, P = 0.000]. Furthermore, blockade with anti-CXCR4 antibody before rRIPoC markedly decreased therapeutic efficacy[rRIPoC （2.33 ± 0.46） %, CXCR4-Ab （1.82 ± 0.36） %, n = 10,

关 键 词：骨髓 间质干细胞移植 心肌梗死 心肌缺血 

分 类 号：R542.2[医药卫生—心血管疾病]