Analysis Of The Expression Profile Of Non-coding RNA In Uva-induced Photoaging Of Human Fibroblasts

Objective: Repeatedly irradiate fibroblasts with long-wave ultraviolet(UVA)to establish a photoaging model to explore the molecular mechanism of long-wave ultraviolet(UVA)on the photoaging of human skin fibroblasts(HSF).Methods:Take the foreskin tissue of healthy men aged 20-40,trypsin digestion method to isolate dermal fibroblasts,take the 3rd to 8th generation fibroblasts for follow-up experiments,using 0 J/cm2,5 J/cm2,7.5 J/cm2 10J/cm2 dose of UVA was irradiated to HSF respectively,and fibroblasts were observed 48 h after irradiation,and the proliferation activity of the cells was tested by CCK8 technology.The skin fibroblasts were divided into 7 groups,A: blank control group;B: UVA irradiation control group;Group C: 0.01umol/L genistein group;Group D 0.1umol/L genistein group;E: 1umol/L genistein group;F: 10umol/L genistein group;G: 100 umol/L dye Lignin group,B,C,D,E,F,G groups were all irradiated with UVA.CCK8 technology to determine the effect of genistein on the proliferation of fibroblasts;Western-blot to determine the expression changes of the aging-related protein p21 in the blank,irradiated,and genistein groups;small RNA sequencing technology to detect the photoaging model Use the miranda,targetscan,and starbase databases to predict the differentially expressed microRNA target genes,perform GO and KEGG enrichment analysis on the predicted target genes,and screen out the core microRNAs and target genes regulated by microRNAs,Analyze the results.Results: After UVA irradiation of 0,5,7.5,10J/cm2,the proliferation ability of HSF under different light intensities was detected with CCK8 reagent.The HSF in the UVA treatment group showed signs of aging,and the cell proliferation ability was lower than before(P< 0.05).Compared with group B,the proliferation activity of CF group was higher,and the difference was statistically significant(P< 0.05).After 5J/cm2 UVA irradiation,more than 70% of fibroblasts remained viable.According to previous literature reports,this study selected 5J/cm2 UVA to establish a photoaging model,which was irradiated once a day at a fixed time for a total of 5 days.48 h after irradiation,the expression of cell senescence-related protein p21 was increased compared with the blank control group.Seven microRNAs were screened based on the significance of the difference: hsa-mi R-1290,hsa-mi R-1307-5p,hsa-hsa-mi R-663 a,hsa-mi R-10396a-5p,hsa-mi R-3195,hsa-mi R-9901,hsa-mi R-10401-5p.Using bioinformatics technology to analyze the selected microRNAs,mi Randa,targetscan,and starbase database predict some of the target genes of differentially expressed microRNAs,suggesting that the differentially expressed microRNAs may be associated with axonogenesis,embryonic organ development,calmodulin binding,and metal ions Related to transmembrane transport protein activity.Conclusion:UVA irradiation can cause photoaging of fibroblasts.A certain concentration of genistein(0.01umol/L-10umol/L)has a protective effect on photoaging of fibroblasts.Differentially expressed microRNAs can be regulated by acting on target genes.The photoaging process of cells.