骨髓间充质干细胞来源外泌体诱导自噬对MPP+抑制SH-SY5Y细胞存活的影响及其机制

doi:10.13481/j.1671-587X.20220308

Abstract

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^－1MPP⁺+BMSCs-Sup），0.50 mmol·L^－1 MPP⁺+BMSCs-Exo group （given 0.50 mmol·L^－1 MPP⁺+100 mg·L^－1BMSCs-Exo），MPP⁺+rapamycin（Rap） group （given 0.50 mmol·L^－1MPP⁺+2 mmol·L^－1 Rap） and MPP⁺+BMSCs-Exo+3-methyladenine（3-MA） group（given 0.50 mmol·L^－1 MPP⁺+100 mg·L^－1BMSCs-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

Wentao WANG,Xuguang MI,Yang ZHOU,Wenxing PU,Jiaxu GAO,Meng JING,Fankai MENG. 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[J].Journal of Jilin University(Medicine Edition), 2022, 48(3): 606-614.

Figures/Tables 15

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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

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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