过表达吲哚胺2,3双加氧酶大鼠骨髓间充质干细胞通过外排体抑制免疫排异反应机制的研究

发布时间：2018-06-10 08:10

本文选题：大鼠骨髓间充质干细胞 + 吲哚胺　；参考：《临床心血管病杂志》2017年11期

【摘要】：目的:探讨过表达吲哚胺2,3双加氧酶(IDO)大鼠骨髓间充质干细胞(BMSCs)通过分泌外排体(exosome)抑制免疫排异反应机制研究。方法:通过慢病毒载体GV308携带IDO转染干细胞构建过表达IDOBMSCs体系并加入基因开启剂强力霉素(DOX)后采用SBI公司的ExoQuick-TC提取分泌的exosome。同时建立大鼠腹腔异位移植心脏模型,经尾静脉给予相应细胞分泌的exosome,利用超声心动图检测经注射过表达IDOBMSCs exosome(过表达IDO-BMSCs exosome组,A组)、空载体-BMSCs exosome(空载体-BMSCs exosome组,B组)、BMSCs exosome(BMSCs exosome组,C组)48h后移植心脏心功能变化,将移植未处理组、给予吗替麦考组作为对照。完成心脏彩超后取A组、B组、C组大鼠注射48h后的脾脏,采用流式细胞技术检测其CD40、CD86、CD80、MHCII、CD274、CD45RA、CD45RA+CD45RB、Treg细胞的表达情况,将移植未处理组、给予吗替麦考组及正常大鼠(正常组)脾脏细胞作为对照。进一步采用TMT标记定量蛋白质组学分析A组有关调节免疫排异的蛋白。结果:A组的LVEF、FS较其他各组有提高;采用流式细胞技术证实其CD40、CD86、CD80、MHCII、CD45RA、CD45RA+CD45RB表达降低,而CD274、Treg细胞的表达增高。蛋白质谱示:Four and a half LIM domains 1蛋白为调节免疫排异反应的关键蛋白。结论:采用TMT标记过表达IDO-BMSCs分泌exosome内蛋白,Four and a half LIM domains 1蛋白为调节免疫排异反应的关键蛋白。
[Abstract]:Aim: to investigate the mechanism of inhibition of immune rejection by secreting exosome of rat bone marrow mesenchymal stem cells (BMSCs) overexpression of indoleamine 2nd3 dioxygenase (IDO). Methods: Lentivirus vector GV308 carrying IDO transfected stem cells was used to construct the system of expressing IDOBMSCs and adding doxycycline (DOX) to extract exosome-secreted by ExoQuick-TC of SBI Company. At the same time, an ectopic heart transplantation model was established in rats. Exosome secreted by the corresponding cells was given via caudal vein. Echocardiography was used to detect the cardiac function changes of IDO-BMSCs exosome-expressing IDO-BMSCs exosome group (group A) and empty vector -BMSCs exosome group (empty vector -BMSCs exosome group) BMSCs exosome group B group (BMSCs exosome group C group) 48 hours after transplantation. The untreated group was given Matimex as a control group. The spleen of rats in group A, B and C were injected 48 hours after echocardiography, and the expression of CD40, CD86, MHCII, CD274RA45RA45RA, CD45RBT, Treg cells were detected by flow cytometry, and the untreated group was transplanted into the untreated group, and the expression of CD40, CD86, MHCII, CD274and CD45RA-CD45RA, CD45RBT / Treg cells were detected by flow cytometry. The spleen cells of the Mutimex group and the normal rats (normal group) were given as control group. Furthermore, TMT labeled quantitative proteomics was used to analyze group A proteins involved in regulation of immune rejection. Results compared with other groups, the expression of CD40, CD86, MHCI, CD45RA, CD45RB and CD274Treg cells were decreased and CD274Treg cells were increased by flow cytometry. The protein profile showed that the 1: four and a half Lim domains 1 protein was the key protein to regulate immune rejection. Conclusion: the overexpression of exosome secreted by IDO-BMSCs with TMT-labeled expression of four and a half Lim domains 1 protein is the key protein to regulate immune rejection.
【作者单位】：云南省第一人民医院心脏大血管外科;
【基金】：国家自然科学基金项目(No:81460073,31460298) 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】：R392

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