Bone marrow-derived mesenchymal stem cells overexpressing MiR-21 efficiently repair myocardial damage in rats.
Bone marrow-derived mesenchymal stem cells overexpressing MiR-21 efficiently repair myocardial damage in rats.
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过度表达 MiR-21 的骨髓间充质干细胞可有效修复大鼠心肌损伤
DOI：
10.18632/oncotarget.16254
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发表时间：
2017-04-25
期刊：
影响因子：
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通讯作者：
Yang T
中科院分区：
文献类型：
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作者：
Zeng YL;Zheng H;Chen QR;Yuan XH;Ren JH;Luo XF;Chen P;Lin ZY;Chen SZ;Wu XQ;Xiao M;Chen YQ;Chen ZZ;Hu JD;Yang T
我们研究了过表达微小RNA - 21（miR - 21）的骨髓间充质干细胞（BMSCs）修复蒽环类药物诱导的大鼠心脏损伤的能力。
选取2 - 3周龄的Sprague - Dawley（SD）大鼠分离培养BMSCs。构建携带pLVX - miR - 21的慢病毒并转染大鼠BMSCs。将大鼠分为未处理的阴性对照组，以及单独注射阿霉素组，或先注射阿霉素后分别注射BMSCs、pLVX - BMSCs或pLVX - miR - 21 - BMSCs组（每组n = 10）。通过胆囊收缩素 - 8（CCK - 8）和Transwell检测细胞的增殖和迁移。通过实时定量聚合酶链反应（qRT - PCR）检测miR - 21的表达以及B细胞淋巴瘤2（Bcl2）、BAX（Bcl - 2相关X蛋白）和血管内皮生长因子（VEGF）的mRNA表达。采用蛋白质印迹法检测Bcl - 2、Bax和VEGF的蛋白表达。
通过CCK - 8和Transwell实验，我们发现过表达miR - 21的pLVX - miR - 21 - BMSCs比未转染的BMSCs或pLVX - BMSCs表现出更强的增殖和迁移能力。超声心动图和免疫组化分析表明，在五组中，pLVX - miR - 21 - BMSC组心脏功能改善最明显且血管生成增强。此外，pLVX - miR - 21 - BMSC组Bcl - 2、VEGF和Cx43表达增强，Bax、脑钠肽（BNP）和肌钙蛋白T表达降低。
这些研究结果表明，miR - 21过表达增强了BMSCs的增殖、侵袭和分化能力，以及修复蒽环类药物诱导的心脏损伤和恢复心脏功能所必需的关键因子（Bcl - 2、VEGF和Bax）的表达。
We investigated the ability of bone marrow derived mesenchymal stem cells (BMSCs) overexpressing microRNA-21 (miR-21) to repair cardiac damage induced by anthracyclines in rats. Sprague-Dawley (SD) rats of 2~3 weeks old were selected to isolate and culture BMSCs. A lentivirus harboring pLVX-miR-21 was generated and transfected into rat BMSCs. The rats were assigned into an untreated negative control group, and groups injected with adriamycin alone or with adriamycin followed by BMSCs, pLVX-BMSCs or pLVX-miR-21-BMSCs (n = 10 each). Proliferation and migration of cells were detected by cholecystokinin-8 (CCK- 8) and transwell. MiR-21 expression, mRNA expressions of B cell lymphoma 2 (Bcl2), BAX (BCL-2-associated X protein) and vascular endothelial growth factor (VEGF) were tested by qRT-PCR. Western blotting was applied to detect protein expressions of Bcl-2, Bax and VEGF. Using CCK- 8 and transwell assays, we found that pLVX-miR-21-BMSCs, which overexpressed miR-21, exhibited greater proliferation and migration than untransfected BMSCs or pLVX-BMSCs. Ultrasonic cardiograms and immunohistochemical analysis demonstrated that among the five groups, the pLVX-miR-21-BMSC group exhibited the most improved heart function and enhanced angiogenesis. Moreover, the pLVX-miR-21-BMSC group showed enhanced expression of Bcl-2, VEGF and Cx43 and reduced expression of Bax, BNP and troponin T. These findings suggest miR-21 overexpression enhanced the proliferation, invasiveness and differentiation of BMSCs as well as expression of key factors (Bcl-2, VEGF and Bax) essential for repairing the cardiac damage induced by anthracyclines and restoring heart function.