过表达IDO骨髓间充质干细胞通过外排体促进移植心脏存活的分子机制研究

贺继刚; 韩金秀; 李贝贝; 李宏远; 撒亚莲; 严丹; 李洪荣

doi:10.13201/j.issn.1001-1439.2018.03.021

过表达IDO骨髓间充质干细胞通过外排体促进移植心脏存活的分子机制研究

- 云南省第一人民医院心脏大血管外科(昆明, 653200)
基金项目:
国家自然科学基金（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）

详细信息

通讯作者: 李洪荣,E-mail:jiganghe@sina.com

中图分类号: R654.2

收稿日期: 2017-07-19

Molecular mechanism of the overexpression of IDO bone marrow mesenchymal stem cells for the survival of transplanted heart

- Cardiovascular Surgery, the First People's Hospital of Yunnan Province, Kunming, 650032, China

More Information

Corresponding author: LI Hongrong, E-mail: jiganghe@sina.com

Received Date: 19 July 2017

摘要

摘要: 目的：探讨过表达IDO大鼠骨髓间充质干细胞（BMSCs）通过分泌外排体促进移植心脏存活的分子基础。方法：通过慢病毒载体GV308携带IDO转染构建过表达IDO大鼠BMSCs，并加入基因开启剂强力霉素（DOX），按照分泌外排体的类型分为3组：过表达IDO-BMSCs-exosome组（过表达IDO组）、空载体-BMSCs-exosome组（空载体组）、BMSCs-exosome组（BMSCs组）；然后采用SBI公司的ExoQuick-TC提取3组分泌的外排体，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外排体，利用彩色超声心动图检测注射3组外排体后2 d移植心脏的心功能变化。另将注射吗替麦考（吗替麦考组）及建模未处理（未处理组）的大鼠作为对照。进而采用小RNA测序技术检测过表达IDO组大鼠BMSCs外排体中与免疫相关的microRAN表达。结果：心脏彩色超声结果显示：过表达IDO-BMSCs分泌的外排体可以有效改善异位移植心脏存活。而根据KEGG分析可见前20位上调microRNA中涉及免疫的有10个，其中miR-540-3p的差异倍数（FC）值上升幅度最大。前20位下调microRNA中涉及免疫的有3个，其中miR-338-5p的FC值下降幅度最大。结论：通过采用小RNA （sRNA）测序技术检测过表达IDO组大鼠BMSC分泌外排体内与免疫相关的microRNA，最终确认上调重点microRNA为miR-540-3p，下调重点microRNA为miR-338-5p。
- IDO /
- 外排体 /
- microRNA /
- 骨髓间充质干细胞
Abstract: Objective: To investigate the molecular basis of the overexpression of IDO rat bone marrow mesenchymal stem cells (BMSCs) secreted exosomes contribute to the survival of transplanted heart. Method: Overexpressed IDO rat BMSCs were constructed using lentiviral vectors (GV308) that carry IDO transfected rat BMSCs, which was then put into gene activators (doxycycline). According to the type of exosome, divided into three groups, each containing three samples. Three groups were:overexpression of IDO-BMSCs-exosome, empty vector-BMSCs-exosome, BMSCs-exosome.The exuQuick-TC of SBI was used to extract the secreted exosomes, and the model of ectopic endothelial cardiac transplantation was created. The secreted exosome was injected through the tail vein. Echocardiography was performed to examine function of transplanted heart after a 2-day treatment with exosome. Simultaneous Mycophenolate mofetil group and untreated group as a control group. MicroRAN sequencing technique was applied to examine immunity-related over-expression IDO rat BMSCs in exosome secretion. Result: Doppler ultrasonography showed that the exosome secreted by overexpressed IDO-BMSCs greatly contributed to the survival of ectopic transplanted heart. KEGG analysis showed that among the top 20 up-regulated microRNAs, 10 were involved in immunity. Furthermore, the FC value of microRNA-540-3p increased in a large margin. And top 20 down-regulated microRNAs, 10 were involved in immunity. Furthermore, the FC value of microRNA-338-5p decreased in a large margin.Conclusion: Overexpressed IDO rat BMSCs are detected during exosome secretion and immunity-related expression of microRNA. MicroRNA-540-3p is up-regulated, whereas microRNA-338-5p is down-regulated.
- indoleamine 2,3-dioxygenase /
- exosome /
- microRNA /
- bone marrow mesenchymal stem cells [LM]

HTML全文

参考文献(15)

[1]	Zaher SS, Germain C, Fu H, et al.3-hydroxykynurenine suppresses CD4⁺ T-cell proliferation, induces T-regulatory-cell development, and prolongs corneal allograft survival[J].Invest J Vis Sci, 2011, 52(5):2640-2648.
[2]	Spielmann N, Wong DT. Saliva:diagnostics and therapeutic perspectives[J].Oral Dis, 2011, 17(4):345-354.
[3]	Lei D, Zhang F, Yao D, et al. MiR-338-5p suppresses proliferation, migration, invasion, and promote apoptosis of glioblastoma cells by directly targeting EFEMP1[J]. Biomed Pharmacother, 2017, 89:957-965.
[4]	Chen X, Pan M, Han L, et al.miR-338-3p suppresses neuroblastoma proliferation, invasion and migration through targeting PREX2a[J].FEBS Lett, 2013, 587(22):3729-3737.
[5]	Na H, Wu Z, Li L, et al. MiR-338-3p inhibits epithelial-mesenchymal transition in gastric cancer cells by targeting ZEB2 and MACC1/Met/Akt signaling[J]. Oncotarget, 2015, 6(17):15222-15234.
[6]	Sun J, Feng X, Gao S, et al. microRNA-338-3p functions as a tumor suppressor in human non-small-cell lung carcinoma and targets Ras-related protein 14[J]. Mol Med Rep, 2015, 11(2):1400-1406.
[7]	Xing Z, Lan Y, Xian L, et al. Anticancer bioactive peptide-3 inhibits human gastric cancer growth by targeting miR-338-5p[J]. Cell Biosci, 2016, 6(1):53.
[8]	Leonard WJ, O'Shea JJ. Jaks and STATs:biological implications[J]. Annu Rev Immunol, 1998, 16(16):293-322.
[9]	Mishra J, Karanki SS, Kumar N. Identification of molecular switch regulating interactions of Janus kinase 3 with cytoskeletal proteins[J]. J Biol Chem, 2012, 287(49):41386-41391.
[10]	Mishra J, Waters CM, Kumar N. Molecular mechanism of interleukin-2-induced mucosal homeostasis[J]. Am J Physiol Cell Physiol, 2012, 302(5):735-747.
[11]	Kumar N, Mishra J, Narang VS, et al. Janus kinase 3 regulates interleukin 2-induced mucosal wound repair through tyrosine phosphorylation of villin[J]. J Biol Chem, 2007, 282(42):30341-30345.
[12]	Johnston JA, Kawamura M, Kirken RA, et al. Phosphorylation and activation of the Jak-3 Janus kinase in response to interleukin-2[J]. Nature, 1994, 370(6485):151-153.
[13]	Fujimoto M, Naka T, Nakagawa R, et al. Defective thymocyte development and perturbed homeostasis of T cells in STAT-induced STAT inhibitor-1/suppressors of cytokine signaling-1 transgenic mice[J]. J Immunol, 2000, 165(4):1799-1806.
[14]	Degryse S, de Bock CE, Cox L, et al. JAK3 mutants transform hematopoietic cells through JAK1 activation, causing T-cell acute lymphoblastic leukemia in a mouse model[J]. Blood, 2014, 124(20):3092-3100.
[15]	Henkels KM, Frondorf K, Gonzalezmejia ME, et al. IL-8-induced neutrophil chemotaxis is mediated by janus kinase 3(jak3)[J]. FEBS Lett, 2011, 585(1):159-166.