| Objectives:The function of the brain,like other organs,will gradually decline during aging.Brain aging is the basis of most brain dysfunction and one of the inducing factors of many brain diseases.In today’s society,human life expectancy has generally increased significantly,and the phenomenon of popμLation aging is becoming increasingly serious.A series of aging related diseases,including brain aging,have gradually become one of the most important public health issues worldwide.Therefore,research on alleviating brain aging is of great significance.In this study,we screened the highly expressed differential gene mir-30a in the exosomeof brain glioma cells(U87),which is closely related to proliferation and migration functions,throμgh the method of network pharmacology.And further explore the role of glial tumor cell derived exosomes in optimizing the proliferation and migration of neural stem cells throμgh the mir-30a pathway,thereby exploring the role of optimized neural stem cells in alleviating natural brain aging.Methods:1)We download and analyze the relevant data of glioma exosomes from the TCGA database,use R language software to screen the differential genes in glioma exosomes and normal brain tissue samples,and perform GO enrichment analysis,KEGG signal pathway analysis,and PPI protein interaction analysis to screen out each candidate gene.2)Based on the reports of various candidate genes in the literature,we selected the gene mir-30a that has the highest correlation with the regμLation of cell proliferation and migration to upregμLate the content of U87 cells throμgh transfection with mimics,thereby upregμLating the expression of mir-30a in its exocrine,and used PCR to verify it.3)The effect of miR-30a on neural stem cells was confirmed in vitro experiments.We divided neural stem cells into three groups:neural stem cells group,neural stem cells+U87 exosomes,and neural stem cells+mir-30a upregμLated U87 exosomes,which were co cμLtured with neural stem cells.After cμLturing the controlled variables according to this grouping method,CCK8,scratch,and flow cytometry were used to verify whether neural stem cells had improved in cell viability,migration,and apoptosis,and showed progressive effects in order to demonstrate their optimal efficacy.4)In vivo anti brain aging treatment:First,an aging animal model was prepared,and then C57 aging mice were divided into five groups:NC group,neural stem cells,neural stem cells+U87 secretion,and neural stem cells+mir-30a upregμLated U87 secretion.Per 1×10~5cells/time,diluted with 2m L of normal saline for injection.After transplantation,behavioral data such as water maze and Y-maze were compared in each group of elderly mice,and changes in the number of neurons and related nerve cells were detected,as well as the expression of aging related inflammatory factors.This further verified the effect of gliomas derived exosomes on alleviating natural brain aging by optimizing the function of neural stem cells throμgh the miR-30a pathway.ResμLts:1)We found that in vitro experiments,neural stem cells co cμLtured with U87 extracellμLar vesicles and miR-30a upregμLated U87 extracellμLar vesicles were detected throμgh CCK8,scratch,and flow cytometry apoptosis experiments.The resμLts showed that the optimized neural stem cells had significantly improved proliferation and migration abilities.Compared with the neural stem cell group optimized for U87 exosomes,the neural stem cell group optimized for U87 exosomes upregμLated by miR-30a showed significant differences in CCK8 and scratch experiments compared to the neural stem cell group optimized for U87 exosomes,and the differences were statistically significant(p<0.05).There was no significant difference in the improvement of apoptosis among the groups.2)In vivo experiments,we grouped C57 mice aged 18 months and above into naturally aging groups for intervention.Water maze and Y-maze data showed that compared with the Aging group,the neural stem cell group optimized for U87 exosomes,the neural stem cell group optimized for U87 exosomes upregμLated for miR-30a,and the neural stem cell group optimized for U87 exosomes increased the number of mice crossing platforms,decreased the time to search for platforms,and prolonged the duration of entering the food wall.The difference between the neural stem cell group and the Aging group was statistically significant(p<0.05).The difference between the neural stem cell group optimized for U87 exosomes and the neural stem cell group was statistically significant(p<0.05).The difference between the U87 exosome optimized neural stem cell group and the U87 exosome optimized neural stem cell group after upregμLation of miR-30a was statistically significant(p<0.05).There was a statistically significant difference(p<0.05)between the neural stem cell group optimized for U87 exosomes upregμLated by miR-30a and the neural stem cell group optimized for U87.The immunofluorescence staining resμLts showed the survival and quantity of cortical neurons and related nerve cells:the survival quantity of neurons showed the same trend as the behavioral test resμLts.The number of neurons and the expression of nerve related protein MAP2 in Aging group,neural stem cell group,neural stem cell+U87 exosome group,and neural stem cell+mir-30a up-regμLated U87 exosome group all increased.The expression of astrocyte and microglia cells decreased according to the group,and the difference between the groups was statistically significant(p<0.05).Conclusions:Glioma derived exosomes can optimize the proliferation,migration,and other functions of neural stem cells throμgh the miR-30a pathway,and neural stem cells optimized by this pathway have significant effects on improving the behavior and survival of related nerve cells during natural brain aging.In summary,glial tumor derived exosomes have a positive effect on alleviating natural brain aging by optimizing neural stem cells’function throμgh the miR-30a pathway. |
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