Study on the paracrine effects of LED yellow light irradiated human mesenchymal stem cells on fibroblast photoaging
Objective To investigate the impact of paracrine effects of LED yellow light irradiated human adipose derived mesenchymal stem cells(hADSCs)on human dermal fibroblasts(HDFs)photoaging.Methods A photoaging model was established by irradiating HDFs with medium-wavelength ultraviolet(UVB)light,followed by assays to evaluate cell proliferation,senescence,and migration.Cell proliferation assays were conducted on hADSCs exposed to LED yellow light with varying energy gradients(2.5,5,7.5,10 J/cm2)to determine the optimal irradiation energy.In the experimental group,HDFs subjected to photoaging were cultured with the supernatant from hADSCs irradiated with LED yellow light at 10 J/cm2.In the control group,HDFs were cultured with the supernatant from non-irradiated hADSCs.Cell proliferation was assessed using the CCK-8 assay,cellular senescence levels were evaluated by β-galactosidase staining,the cytoskeleton was labeled with green fluorescent probes targeting actin filaments,while reactive oxygen species(ROS)content was measured by flow cytometry.Additionally,transcriptome sequencing was performed,and significant differentially expressed genes identified through sequencing were validated by quantitative real-time PCR(qRT-PCR).The KEGG pathway enrichment analysis was also conducted for the differentially expressed genes.Results Compared to the non-irradiated group,the UVB-irradiated group showed a significant decrease in HDFs proliferation,a marked increase in senescent cells(blue-stained cells),and inhibited HDFs migration.All differences were statistically significant(P＜0.05).Compared to the control group,HDFs in the experimental group treated with the supernatant from LED yellow light irradiated hADSCs showed enhanced proliferation,fewer blue-stained senescent cells,a more elongated cell morphology,and lower ROS levels.All differences were statistically significant(P＜0.05).Transcriptome sequencing identified six significantly differentially expressed genes,including Kinesin family member 20B(KIF20B),Abnormal spindle-like microcephaly associated protein(ASPM),A-kinase anchor protein 9(AKAP9),Centromere protein F(CENPF),Centromere protein E(CENPE),and RB1-induced coiled-coil protein 1(RB1CC1).qRT-PCR results showed that the mRNA expression levels of KIF20B,ASPM,AKAP9,CENPF,CENPE,and RB1CC1 were higher in the experimental group compared to the control group,with all differences being statistically significant(P＜0.05).These results were consistent with the transcriptome sequencing data.KEGG pathway enrichment analysis revealed that the significantly differentially expressed genes were primarily enriched in the longevity regulating pathway.Conclusion The paracrine effects of LED yellow light irradiated hADSCs exhibited an inhibitory effect on HDFs photoaging,which may be related to the activation of the longevity regulating pathway.
万方数据
维普
