Chinese Journal of Tissue Engineering Research ›› 2019, Vol. 23 ›› Issue (29): 4617-4622.doi: 10.3969/j.issn.2095-4344.1809

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Time-volume variation in miR-330-3p expression in GATA-4-overexpressing bone marrow-derived mesenchymal stem cell exosomes 

He Jigang1, Xie Qiaoli1, Wang Zihao1, Yan Dan2, Zhang Hongbo2   

  1. 1Department of Cardiovascular Surgery, 2Department of Intensive Care Unit, Affiliated Hospital of Kunming University of Science and Technology, the First People’s Hospital of Yunnan Province, Kunming 650032, Yunnan Province, China
  • Revised:2019-04-12 Online:2019-10-18 Published:2019-10-18
  • Contact: Zhang Hongbo, Attending physician, Department of Intensive Care Unit, Affiliated Hospital of Kunming University of Science and Technology, the First People’s Hospital of Yunnan Province, Kunming 650032, Yunnan Province, China
  • About author:He Jigang, MD, Associate chief physician, Department of Cardiovascular Surgery, Affiliated Hospital of Kunming University of Science and Technology, the First People’s Hospital of Yunnan Province, Kunming 650032, Yunnan Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81460073 (to HJG); Yunnan Provincial Department of Science and Technology - Kunming Medical University Joint Research Project, No. 2014FB089 (to HJG); Yunnan Provincial Department of Education Science Research Fund, No. 2015Z051 (to HJG); China Postdoctoral Science Foundation, No. 2015M582764XB (to HJG); 2015 Research Project of Chengdu Medical College, No. CYZ15-18 (to HJG); Yunnan Medical Reserve Talents, No. H-201607 (to HJG)

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Abstract:

BACKGROUND: Previous experiments have demonstrated that exosomes secreted by bone marrow mesenchymal stem cells overexpressing GATA-4 (BMSCsGATA-4) can effectively repair myocardial injury induced by myocardial infarction through inhibiting cardiomyocyte apoptosis. We further discovered that miRNA-330-3p is highly expressed in BMSCsGATA-4 exosomes and involved in anti-apoptosis, suggesting that miRNA-330-3p may be a key molecule for exosomes to repair myocardial injury.
OBJECTIVE: To investigate the time-volume variation in miR-330-3p expression in BMSCsGATA-4 exosomes. 
METHODS: Experimental group 1 (BMSCsGATA-4–miR-330-3p-mimic) and experimental group 2 (BMSCsGATA-4–miR-330-3p-inhibitor) were established by adding miR-330-3p-mimic or miR-330-3p-inhibitor to the culture systems of BMSCsGATA-4 exosomes, respectively. The BMSCsGATA-4, BMSCsGATA-4–empty vector, BMSCs, BMSCsGATA-4–miR-330-3p-mimic–empty vector, and BMSCsGATA-4–miR-330-3p-inhibitor–empty vector groups were taken as confounding factors and incubated for 24 or 48 hours with doxycycline, a gene opener. Exosomes secreted by each group were extracted. RT-PCR was used to detect the expression level of miR-330-3p in each group of cells and secreted exosomes. Morphological changes in the BMSCsGATA-4-microRNA-330-3p-mimic, BMSCsGATA-4-microRNA-330-3p-inhibitor, and BMSCs groups at 24 and 48 hours of incubation with doxycycline were observed under light microscopy. 
RESULTS AND CONCLUSION: The expression of miR-330-3p was highest in the cells and exosomes of the BMSCsGATA-4–miR-330-3p-mimic group (P < 0.05), and increased gradually with time. Conversely, the expression of miR-330-3p was lowest in the cells exosomes of the BMSCsGATA-4-miR-330-3p-inhibitor group, and moreover, it decreased gradually with time. There were no morphological changes in the BMSCsGATA-4–miR-330-3p-mimic, BMSCsGATA-4–miR-330-3p-inhibitor, and BMSC group at 24 and 48 hours of incubation with doxycycline. Therefore, miR-330-3p expression in BMSCs is positively correlated with that in exosomes. Expression of miR-330-3p in exosomes can be effectively increased through miR-330-3p overexpression while reduced by silencing.

Key words: myocardial infarction, myocardial injury, bone marrow mesenchymal stem cells, GATA-4, exosomes, miR-330-3p, National Natural Science Foundation of China

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