Screening method for endogenous aging targets of skins, active substance capable of improving endogenous aging of skins and screening method thereof
A screening method, an in-skin technology, applied to the screening method of skin endogenous aging targets, the active substances that improve skin endogenous aging, the field of screening active substances with targets, can solve the function and target are not fully explained or Confirmation and other issues to achieve the effect of high specificity and high sensitivity
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Embodiment 1
[0050] Example 1: Detection of expression profiles of high-throughput microRNA chips in young skin samples and aging skin samples
[0051] 1. Obtaining of skin samples
[0052] Fresh facial skin samples were purchased from Shanghai Xinchao Biotechnology Co., Ltd. Young skin sample donors were healthy women aged between 30 and 40 who were born in Shanghai and lived in Shanghai. There were 8 cases in total. The average age of this group was They were (33.5±2.6) years old; donors of aging skin samples were healthy women who were older than 50 years old, born and lived in Shanghai, a total of 7 cases, and the average age of this group was (62.0±9.7) years old.
[0053] Specifically, the skin tissue is collected after conventional plastic surgery, and the skin tissue is processed as soon as possible. Cut the skin tissue, the thickness of the skin does not exceed 0.5cm, cut into small tissue pieces of 0.5cm x 1cm x 1cm (thickness x length x width), and then put these small tissue p...
Embodiment 2
[0058] Example 2: Screening and induction of differential microRNAs in young skin samples and aging skin samples
[0059] The differential expression of microRNAs in young and aged skin samples was studied using the AgiMicroRna R analysis toolkit, which is based on a limma linear model to process the test data. Analysis results such as figure 1 As shown in the clustering heatmap, there are a total of 17 microRNA molecules with significant differences in expression levels between young skin samples and aged skin samples, of which 12 microRNAs have significantly increased expression levels in aged skin samples, and 5 microRNAs have significantly decreased expression levels microRNA, and the expression differences of these microRNA molecules are shown in Table 1.
[0060] Table 1. A list of microRNA molecules with significant differences between young skin and aging skin screened by microarray experiments (the name of the microRNA molecule compared with the reference of the pres...
Embodiment 3
[0063] Example 3: Bioinformatics analysis to obtain microRNA markers and target genes of skin endogenous aging
[0064] Using the "Validated Target Module" module of the miRWalk 2.0 database to predict the target genes of differential microRNAs in young and aging skin samples, a set of predicted gene sets was generated through this step.
[0065] Functional and metabolic pathway enrichment analyzes were performed on the predicted gene set using the bioinformatics database DAVID v6.8. The enrichment analysis is based on five annotation category modules, including Gene Ontology (GO_TERM), KEGG pathway, BIOCARTA pathway, InterPRO database and UP_KEYWORDS database. Part of the analysis results are shown in Table 2, which lists the five functional or metabolic pathway annotations with the highest enrichment degree.
[0066] Table 2. Functional and metabolic pathway enrichment analysis results of differential microRNA target genes
[0067]
[0068] Among the functions and metab...
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