Journal of Shanghai Jiao Tong University (Medical Science) >

2022 , Vol. 42 >Issue 3: 323 - 330

DOI: https://doi.org/10.3969/j.issn.1674-8115.2022.03.009

Basic research

Effect of miR-124-1 mediated by exosomes of bone marrow-derived mesenchymal stem cells on the regulation of transformation of M2 microglia

  • Lei HAO ,
  • Ge JIN ,
  • Yongtao YANG ,
  • Junwei WANG ,
  • Yang SUN ,
  • Cuiling QIN ,
  • Qunling ZHAN
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  • Department of Neurology, the Fifth People's Hospital of Chongqing, Chongqing 400062, China
ZHAN Qunling, E-mail:aczhan@163.com.

Received date: 2022-01-04

  Online published: 2022-05-09

Supported by

Natural Science Foundation of Chongqing(cstc2020jcyj-msxmX0988)

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Abstract

Objective

·To investigate the effect of exosomes (Exo) of bone marrow-derived mesenchymal stem cells (BMMSCs) overexpressing miR-124-1 gene on the regulation of transformation of M2 microglia (MG).

Methods

·BMMSCs of rats were isolated and cultured. BMMSCs-Exo were extracted. BMMSCs and BMMSCs-Exo were identified by flow cytometry, transmission electron microscope (TEM) and Western blotting, respectively. MiR-124-1 gene was synthesized and its lentiviral vector was constructed. The expression changes of miR-124-1 gene in BMMSCs and BMMSCs-Exo were observed. BMMSCs-Exo overexpressing miR-124-1 gene (Exo/124-1) was co-cultured with HAPI microglia cell line activated by lipopolysaccharide (LPS). Cells from Exo group and EXO/124-1 group were collected, respectively. HAPI cells cultured with LPS only (LPS group) were compared with untreated HAPI cells (normal group). Quantitative real-time PCR (qPCR) and Western blotting were used to detect the mRNA and protein expression of M1 molecules [interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α)] and M2 molecules(CD206, IL-10) in HAPI cells.

Results

·BMMSCs and BMMSCs-Exo were successfully isolated, cultured and identified. Exo/124-1 significantly expressed miR-124-1 gene. BMMSCs-Exo could carry miR-124-1 gene, significantly down-regulating the expression of IL-6 (compared with the normal group and the LPS group, the down-regulation percentage were 55.71% and 73.04%, respectively at the gene level, and the percentages were 52.32% and 76.95%, respectively at the protein level) and TNF-α (compared with the normal group and the LPS group, the down-regulation percentage were 51.95% and 68.91%, respectively at the gene level, and 52.14% and 77.47%, respectively at the protein level) of HAPI cells tested by qPCR and Western blotting. Exo/124-1 up-regulated the expression of CD206 (compared with the normal group and the LPS group, the down-regulation percentage were 46.90% and 76.99%, respectively at the gene level, and 65.13% and 85.11%, respectively at the protein level) and IL-10 (compared with the normal group and the LPS group, the down-regulation percentage were 43.09% and 72.36%, respectively at the gene level, and 55.19% and 78.18%, respectively at the protein level) in HAPI cells.

Conclusion

·BMMSCs-Exo can mediate miR-124-1 to regulate the polarization of HAPI cells to M2 type.

Key words： bone marrow-derived mesenchymal stem cell (BMMSC); exosome (Exo); miR-124-1 gene; HAPI cell; the polarization of M2 type

Cite this article

Lei HAO , Ge JIN , Yongtao YANG , Junwei WANG , Yang SUN , Cuiling QIN , Qunling ZHAN . Effect of miR-124-1 mediated by exosomes of bone marrow-derived mesenchymal stem cells on the regulation of transformation of M2 microglia[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2022 , 42(3) : 323 -330 . DOI: 10.3969/j.issn.1674-8115.2022.03.009

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