过表达GATA-4骨髓间充质干细胞通过外泌体修复心肌损伤的探讨

发布时间：2018-06-14 21:58

本文选题：GATA- + 外排体　；参考：《临床心血管病杂志》2017年09期

【摘要】：目的:探讨过表达GATA-4小鼠骨髓间充质干细胞(BMSC)通过分泌外排体(exosome)修复心肌损伤的机制。方法:通过慢病毒载体GV308携带GATA-4转染小鼠BMSC构建过表达GATA-4小鼠BMSC并加入基因开启剂——强力霉素(DOX),然后采用SBI公司的ExoQuick-TC提取分泌的exosome。通过电镜检测exosome的形态。经尾静脉注射采用Dir预染的过表达GATA-4-BMSCs分泌的exosome(过表达GATA-4组)、空载体-BMSCs分泌的exosome(空载体组)、BMSCs细胞分泌的exosome(BMSCs组),将不给予任何处理的心肌梗死(心梗)模型小鼠及正常喂养的小鼠分别设为未处理组和对照组。采用心脏彩超检测给予干预措施96h的心功能改善情况。进而采用小动物活体成像检测心梗局部Dir荧光强度。番茄凝集素评估心梗局部血管生成及采用免疫组化检测心梗局部C-kit阳性细胞的数量。结果:过表达GATA-4组较其他组可以更好地改善心梗后的心功能,射血分数及环比收缩改善幅度最大。过表达GATA-4组较其他组在96h时心脏局部可见更高荧光强度。番茄凝集素实验可见:过表达GATA-4组较其他组在给予干预措施后96h,可以表达出更多的血管。C-kit免疫组织化学检测可见:过表达GATA-4组较其他组在给予干预措施后96h,心梗局部C-kit细胞数量较其他组多。结论:过表达GATA-4的BMSCs可以通过分泌的exosome增强"归巢"效应、促进心梗局部血管增生、有效动员C-kit阳性细胞修复心肌损伤。
[Abstract]:Aim: to investigate the mechanism of repairing myocardial injury by overexpression of bone marrow mesenchymal stem cells (BMSC) of GATA-4 mice by exosome-secreting exosome. Methods: Lentivirus vector GV308 carrying GATA-4 was used to transfect mouse BMSC to construct the overexpression of GATA-4 mouse BMSC and to add doxycycline, a gene opener, to extract exosome-secreted by ExoQuick-TC of SBI Company. The morphology of exosome was examined by electron microscope. Tail vein injection of exosome secreted by Dir prestained overexpression GATA-4-BMSCs (overexpression of GATA-4 group), exosome-secreted by empty vector -BMSCs (empty vector group) and exosome-BMSCs secreted by BMSCs cells, will be given to myocardial infarction (MI) model mice and positive mice without any treatment. The mice were divided into untreated group and control group. Cardiac function was improved after 96 hours of intervention by echocardiography. Then the local Dir fluorescence intensity of myocardial infarction was detected by small animal in vivo imaging. Tomato lectin was used to evaluate angiogenesis and immunohistochemistry was used to detect the number of C-kit positive cells in myocardial infarction. Results: overexpression of GATA-4 could improve the cardiac function after myocardial infarction, and the ejection fraction and annular contraction were the largest. Overexpression of GATA-4 was more intense in the heart than that in the other groups at 96 h. Tomato agglutinin assay showed that GATA-4 overexpression could express more blood vessel. C-kit immunohistochemistry than other groups at 96h after intervention. It can be seen that GATA-4 overexpression is more effective than other groups in myocardial infarction 96 hours after intervention. The number of local C-kit cells was more than that of other groups. Conclusion: overexpression of GATA-4 can enhance homing effect by secreting exosome, promote regional angiogenesis of myocardial infarction, and mobilize C-kit positive cells to repair myocardial injury.
【作者单位】：云南省第一人民医院心脏大血管外科;
【基金】：国家自然科学基金(No:81460073) 云南省科技厅-昆明医科大学应用基础研究联合专项(No:2014FB089) 云南省教育厅科学研究基金(No:2015Z051) 中国博士后科学基金(No:2015M582764XB) 成都医学院2015年度科研项目(No:CYZ15-18) 云南省医学后备人才(No:H-201607)
【分类号】：R542.2

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