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SDF-1、CXCR4轴在缺氧预处理骨髓间充质干细胞移植促进SD大鼠急性心肌梗死心脏功能恢复中的作用
An Experimental Study on the Expression of SDF-1，CXCR4 in the Acute Infarct Myocardium in SD Rat after Implantation of Hypoxia-preconditioned Bone Marrow MSCs

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DOI:
作者:
盛瑾，夏宇，石蓓，许官学，赵然尊，沈长银，王冬梅，刘志江
Sheng Jin，Xia Yu,Shi Bei,Xu Guanxue,Zhao Ranzun, Sheng Changyin, Wang Dongmei, Liu Zhijiang
作者单位:
贵州省遵义医学院第一附属医院心内科，贵州省遵义医学院第一附属医院心胸外科，贵州省遵义医学院第一附属医院心内科，贵州省遵义医学院第一附属医院心内科，贵州省遵义医学院第一附属医院心内科，贵州省遵义医学院第一附属医院心内科，贵州省遵义医学院第一附属医院心内科，贵州省遵义医学院第一附属医院心内科
Department of Cardiology, Affiliated Hospital of Zunyi Medical College；Department of Thoracic cardiovascular surgery, Affiliated Hospital of Zunyi Medical College；Department of Cardiology, Affiliated Hospital of Zunyi Medical College；Department of Cardiology, Affiliated Hospital of Zunyi Medical College；Department of Cardiology, Affiliated Hospital of Zunyi Medical College；Department of Cardiology, Affiliated Hospital of Zunyi Medical College；Department of Cardiology, Affiliated Hospital of Zunyi Medical College；Department of Cardiology, Affiliated Hospital of Zunyi Medical College
关键词:
骨髓间充质干细胞，基质细胞衍生因子-1，CXCR4，缺氧预处理，心肌梗死 SD大鼠
mesenchymal stem cells， stromal cells-derived factor 1, CXC chemokine receptor 4, hypoxic preconditioning, myocardial infarction
摘要:
目的 观察缺氧预处理的同种异体骨髓间充质干细胞（MSCs）移植入受体后，基质细胞衍生因子SDF-1 CXCR4轴在促进SD大鼠急性心肌梗死心脏功能恢复中的作用。 方法 对SD大鼠MSCs进行体外培养、鉴定、缺氧预处理。128只SD大鼠，随机分为缺氧预处理MSCs移植组（预处理MSCs组，n=32）、MSCs移植组（MSCs组，n=32 ）、单纯心梗对照组（对照组，n=32），正常对照组（假手术组，n=32）。预处理MSCs组结扎冠SD大鼠状动脉左前降支造成急性心肌梗死，10分钟后，在梗死区域、梗死区边缘局部注射共约0.5×106uL- 1个经缺氧预处理的MSCs。MSCs组注射约0.5×106uL- 1个MSCs，对照组注入相同容量生理盐水。其余32只SD大鼠作假手术组。心肌梗死前及梗死后28d采用超声心动图测量LVDd、 LVDs、 EF三项反应心脏功能指标。分别于术后1d、7d、14d和28d每组各处死8只大鼠取心脏组织作免疫组化、常规病理染色、心肌梗死面积、梗死区心室壁厚度和心室肌内微血管密度等项目检查。RT-PCR、免疫激光共聚焦定性及定量测定SDF-1 CXCR4在各组心肌组织内表达变化。 结果 心肌梗死后28d时心肌梗死面积，预处理MSCs组：(18.6±1.88)比(35.0±2.05)，MSCs组：(22.5±2.36)比(34.9±1.91)，对照组：(30.2±2.36)比(35.5±2.76)，组间有显著性差异（P<0.05）。预处理MSCs组、MSCs组、对照组心肌梗死区左室前壁厚度分别为：3.56±0.45mm、2.88±0.32mm、1.80±0.71mm，组间有显著性差异（P0.05）。28d时超声心动图结果，预处理MSCs组LVDd22.85±0.37mm、 LVDs12.98±0.47mm、 EF49.28±5.61，MSCs组LVDd26.23±0.40mm、 LVDs17.00±0.81mm、 EF41.33±6.02，对照组LVDd28.14±0.39mm、 LVDs20.17±0.53mm、 EF33.32±6.88，而假手术LVDd20.99±0.51mm、 LVDs112.99±0.41mm、 EF51.88±4.10，三项反应心脏功能指标预处理MSCs组心脏功能最好，组间比较差异显著（P0.05）。预处理MSCs组心肌梗死边缘区新生微血管量20.97±0.32 （个视野），MSCs组15.77±0.25 （个视野），与对照组5.67±0.15 （个视野）、假手术组8.70±0.63 （个视野），组间比较有显著性差异（P0.05）。预处理MSCs、MSCs、正常心肌组织和梗死心肌组织均有SDF-1、CXCR4表达。移植预处理MSCs和MSCs后SDF-1 CXCR4在梗死周边区域注射细胞处及局部血管壁发生高表达，其中预处理MSCs组更明显。 SDF-1mRNA、CXCR4mRNA、SDF-1和CXCR4蛋白荧光强度在移植后1d、7d、14d、28d时，预处理MSCs组、MSCs组和对照组与假手术组比较均有显著性升高（P0.05），而预处理MSCs组与MSCs组和对照组比较SDF-1mRNA、CXCR4mRNA、SDF-1和CXCR4蛋白荧光强度升高更明显（P0.05）。 结论 SD大鼠急性心肌梗死局部注射缺氧预处理MSCs或MSCs后可使SDF-1、CRCX4在受损组织局部表达上调，同时心功能得到改善。 SDF-1、CRCX4轴在心梗区局部表达上调可能是缺氧预处理MSCs或MSCs移植后改善心脏功能的重要途径之一。
Objective To study the expression of stromal cells-derived factor 1and CXC chemokine receptor 4 (SDF-1，CXCR4) in the acute infarct myocardium in rat after implanation of hypoxic preconditioning bone marrow mesenchymal stem cells (HPMSCs). Methods MSCs were cultured by using the whole bone marrow adherence method. HPMSCs were cultured under hypoxic (0.5 O2 for 24 hrs) before transplantation. SD Rats were anesthetized by intraperitoneal injection of pentobarbital sodium. Lefe anterior thoracotomy through 3, 4 intercostals region was performed, and then the left anterior descending coronary arteries were ligated to produce acute myocardial infarction．The bone marrow mesenchymal stem cells and were injected into the area of acute infarct myocardium after the left anterior descending coronary artery been ligated 10 minutes. The different concentration and expression of SDF-1、CXCR4 in the area of acute myocardial infarction were analyzed. Results The expression of SDF-1 and CXCR4 in topical injection sites of infarct myocardium respectively in the MSCs and HPMSCs implanted groups, were significantly higher than that in infarct site in the control group (p0.05), and HPMSCs group were the highest group in those. The level of SDF-1 and CXCR4 in the HPMSCs implanted group and MSCs implanted groups were highest at 24h after transplantation, then the level began to reduced. Conclusion The MSCs, after being implanted into the acute infarct myocardium, showed the level of SDF-1 and CXCR4 increasing. This study suggested that MSCs and HPMSCs, implanted into the region of acute infarct myocardium, had the ability of increasing of the level of SDF-1 and CXCR4, the effect of HPMSCs were better.

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