The Effect Of Different Sources Of Mesenchymal Stem Cells On Autoimmune Encephalomyelitis Model Mice And Its Mechanism

Background and objectives:Multiple sclerosis is a kind of central nervous system white matter inflammatory demyelination as the main pathological features of autoimmune diseases,the most frequently involved parts of the periventricular white matter,optic nerve,spinal cord and so on.The etiology and pathogenesis is unknown so far,the current view is that innate susceptibility genes carried by an individual,the day after tomorrow the environment,due to the role of viral infection,trauma and other external factors,and disease-induced central component of myelin abnormal autoimmune responseMethods:1.EAE models were prepared by immunizing C57 BL / 6 mice with myelin oligodendrocyte glycoprotein(MOG 35-55)as antigen and complete Freund's adjuvant.The mice were randomly divided into control group,model group,UCMSC group,ADMSC group and UCMSC-SPK1 group.The mesenchymal stem cells from different sources were treated by tail vein,and were injected every seven day after onset.The control group and the model group were treated with the same dose of saline instead,and the neurological deficits were scored on each group of mice by double blindness.2.Each group of mice was subjected to neurological deficit score every day for 32 days.3.Mice were sacrificed on day 33 after immunization,and the brain and spinal cord were removed and stored at-80 ° C.4.The myelination of the spinal cord was observed by electron microscopy5.The expression of GFAP and MBP in the spinal cord of each group was detected by Western blotting.6.The percentage of regulatory T cells(CD4 + CD25 + foxp3 +)and NK(NK1.1 +CD3-)cells in the spleen were detected by flow cytometry.7.The expression of SPK1 in UCMSC,ADMSC and UCMSC-SPK1 was detected by western blot.Results:1.EAE mouse models were successfully prepared by hybrid immunization with MOG35-55 and complete Freund's adjuvant.2.The results of western blot analysis showed that compared with UCMSC,the expression of SPK1 in ADMSC was higher than that in UCMSC,and the expression of SPK1 in UCMSC-SPK1 was higher than that in ADMSC after SPK1 transfection.3.The neurological score of the mice in the UCMSC group,the ADMSC group and the UCMSC-SPK1 group was significantly lower than that in the model group(P<0.05).The scores of the neurological deficits in the mice were significantly lower than those in the control group,Compared with the ADMSC group,the neurological score of UCMSC group was more obvious than that of UCMSC group.Compared with ADMSC group,the trend of neurodegel score in UCMSC-SPK1 group was more significant.4.The spinal cord myelination was observed by electron microscopy.The results showed that the myelin sheath of the spinal cord was lost in the model group,while the myelination area of the three treatment groups was significantly less and the degree was significantly reduced.The differences in the three cell treatment groups were also obvious.Compared with the UCMSC group,the ADMSC group and the UCMSC-SPK1 group,the improvement of the spinal cord was more obvious;compared with the ADMSC group,this trend in the UCMSC-SPK1 group was more Significantly.5.The expression of GFAP in mouse spinal cord were detected by Western blotting.In model group,the expression of GFAP was significantly higher than control group.After cell therapy,the expression of GFAP were more significantly decreased.Compared with the UCMSC group,the expression of GFAP in the ADMSC group and the UCMSC-SPK1 group were significantly lower,and the trend in UCMSC-SPK1 group was lower than that in the ADMSC group.6.The expression of MBP in model group was significantly lower than that in control group,and the expression of MBP in ADMSC group was significantly higher than that of UCMSC group;and this trend in the UCMSC-SPK1 group is more significant.7.The ratio of regulatory T cells and NK cells in the spleen of each group was measured by flow cytometry.The results showed that the proportion of regulatory T cells in the spleen of the model group was significantly lower than that of the control group.The proportion of NK cells in the spleen of UCMSC group,ADMSC group and UCMSC-SPK1 group was significantly increased,and the proportion of NK cells was significantly decreased.Compared with UCMSC group,The ADMSC group and the UCMSC-SPK1 group were more pronounced.Compared with ADMSC group,the expression of regulatory T cells and NK cells in UCMSC-SPK1 group was more significant.Conclusion:1.Different sources of mesenchymal stem cell therapy can significantly reduce the EAE model mice neurological deficit score,the mechanism may be through the regulation of immune response,inhibition of nervous system inflammatory response and promote myelination regeneration to achieve.2.After transfection of SPK1 gene,UCMSC-SPK1 showed better efficacy in the treatment of EAE than ADMSC,while the original UCMSC was less effective than ADMSC.3.Different amounts of stem cells,which express different amounts of SPK1,may lead to differences in their efficacy in the treatment of EAE model mice.