本文選題：GATA- + 外排體　； 參考：《臨床心血管病雜志》2017年09期

【摘要】：目的:探討過表達(dá)GATA-4小鼠骨髓間充質(zhì)干細(xì)胞(BMSC)通過分泌外排體(exosome)修復(fù)心肌損傷的機(jī)制。方法:通過慢病毒載體GV308攜帶GATA-4轉(zhuǎn)染小鼠BMSC構(gòu)建過表達(dá)GATA-4小鼠BMSC并加入基因開啟劑——強(qiáng)力霉素(DOX),然后采用SBI公司的ExoQuick-TC提取分泌的exosome。通過電鏡檢測exosome的形態(tài)。經(jīng)尾靜脈注射采用Dir預(yù)染的過表達(dá)GATA-4-BMSCs分泌的exosome(過表達(dá)GATA-4組)、空載體-BMSCs分泌的exosome(空載體組)、BMSCs細(xì)胞分泌的exosome(BMSCs組),將不給予任何處理的心肌梗死(心梗)模型小鼠及正常喂養(yǎng)的小鼠分別設(shè)為未處理組和對照組。采用心臟彩超檢測給予干預(yù)措施96h的心功能改善情況。進(jìn)而采用小動物活體成像檢測心梗局部Dir熒光強(qiáng)度。番茄凝集素評估心梗局部血管生成及采用免疫組化檢測心梗局部C-kit陽性細(xì)胞的數(shù)量。結(jié)果:過表達(dá)GATA-4組較其他組可以更好地改善心梗后的心功能,射血分?jǐn)?shù)及環(huán)比收縮改善幅度最大。過表達(dá)GATA-4組較其他組在96h時心臟局部可見更高熒光強(qiáng)度。番茄凝集素實(shí)驗(yàn)可見:過表達(dá)GATA-4組較其他組在給予干預(yù)措施后96h,可以表達(dá)出更多的血管。C-kit免疫組織化學(xué)檢測可見:過表達(dá)GATA-4組較其他組在給予干預(yù)措施后96h,心梗局部C-kit細(xì)胞數(shù)量較其他組多。結(jié)論:過表達(dá)GATA-4的BMSCs可以通過分泌的exosome增強(qiáng)"歸巢"效應(yīng)、促進(jìn)心梗局部血管增生、有效動員C-kit陽性細(xì)胞修復(fù)心肌損傷。
[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.
【作者單位】： 云南省第一人民醫(yī)院心臟大血管外科;
【基金】：國家自然科學(xué)基金(No:81460073) 云南省科技廳-昆明醫(yī)科大學(xué)應(yīng)用基礎(chǔ)研究聯(lián)合專項(xiàng)(No:2014FB089) 云南省教育廳科學(xué)研究基金(No:2015Z051) 中國博士后科學(xué)基金(No:2015M582764XB) 成都醫(yī)學(xué)院2015年度科研項(xiàng)目(No:CYZ15-18) 云南省醫(yī)學(xué)后備人才(No:H-201607)
【分類號】：R542.2

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