Journal of Jilin University(Medicine Edition) ›› 2022, Vol. 48 ›› Issue (3): 606-614.doi: 10.13481/j.1671-587X.20220308

• Research in basic medicine • Previous Articles    

Effect of autophagy induced by exosomes derived from bone marrow mesenchymal stem cells on survival of SH-SY5Y cells inhibited by MPP+ and its mechanism

Wentao WANG1,Xuguang MI2,Yang ZHOU1,Wenxing PU1,Jiaxu GAO3,Meng JING4,Fankai MENG4()   

  1. 1.Department of Surgery, Changchun University of Chinese Medicine, Changchun 130117, China
    2.Central Laboratory, People’s Hospital, Jilin Province, Changchun 130021, China
    3.Department of Forensic Medicine, Jilin Shiran Forensic Expertise Center, Changchun 130022, China
    4.Department of Neurosurgery, People’s Hospital, Jilin Province, Changchun 130021, China
  • Received:2021-12-28 Online:2022-05-28 Published:2022-06-21
  • Contact: Fankai MENG E-mail:mfk200372019@126.com

Abstract: Objective

To investigate the effect of autophagy induced by exosomes (exo) derived from the bone marrow mesenchymal stem cells (BMSCs) on the survival of the human neuroblastoma SH-SY5Y cells inhibited by 1-methyl-4-phenylpyridine ion (MPP+), and to darify its mechanism.

Methods

The human neuroblastoma SH-SY5Y cells were given different concentrations(0,0.25,0.50,1.00,and 2.00 mmol·L-1)MMP+,and the survival rates of the SH-SY5Y cells were detected after 24 and 48 h.The experiment was divided into control group[given phosphate buffered saline(PBS)], 0.50 mmol·L-1 MPP+ group (given 0.50 mmol·L-1 MPP+),MPP++BMSCs supernatnt (BMSCs-Sup)group (given 0.50 mmol·L-1 MPP++BMSCs-Sup),0.50 mmol·L-1 MPP++BMSCs-Exo group (given 0.50 mmol·L-1 MPP+ +100 mg·L-1 BMSCs-Exo),MPP+ +rapamycin(Rap) group (given 0.50 mmol·L-1 MPP+ +2 mmol·L-1 Rap) and MPP+ +BMSCs-Exo+3-methyladenine(3-MA) group(given 0.50 mmol·L-1 MPP+ +100 mg·L-1 BMSCs-Exo +1 mmol·L-1 3-MA). The survival rates of the SH-SY5Y cells in various groups were detected by MTT assay.The fluorescence intensities of autophagic acid vesicles in the SH-SY5Y cells in various groups were detected by acridine orange staining under confocal microscope.The expression levels of CD63, TGS101,and Calnexin proteins in the SH-SY5Y cells in various groups were detected by Western blotting method.

Results

The MTT assay results showed that the survival rates of the SH-SY5Y cells were decreased gradually with the increasing of the MPP+ concentration or the extension of treatment time (P<0.05) in a concentration- and time-dependent manner. Compared with MPP+ group, the survival rates of the SH-SY5Y cells in MPP+ + BMSCs-Sup group, MPP+ +BMSCs-Exo group,and MPP+ + Rap group were increased (P<0.05). Compared with MPP+ +BMSCs-Exo group, the survival rate of the SH-SY5Y cells in MPP+ +BMSCs-Exo + 3-MA group was decreased (P<0.05).The acridine orange staining results showed that compared with control group, the fluorescence intensities of autophagic acid vesicles in the SH-SY5Y cells in MPP+ + BMSCs-Sup group, MPP+ +BMSCs-Exo group and MPP+ + Rap group were increased (P<0.05); compared with MPP+ +BMSCs-Exo group, the fluorescence intensity of autophagic acid vesicles in the SH-SY5Y cells in MPP+ +BMSCs-Exo + 3-MA group was decreased (P<0.05).The Western blotting results showed that the exosome marker proteins CD63 and TGS101 were highly expressed, while the endoplasmic reticulum marker protein Calnexin was lowly expressed.

Conclusion

BMSCs-Exo can resist the MPP+-induced decreasing of the survival of the SH-SY5Y cells by activating the autophagy.

Key words: Exosomes, Bone marrow mesenchymal stem cells, Autophagy, Cell viability, SH-SY5Y cells

CLC Number: 

  • R285.5