| Objective:To investigate the differential expressions of microRNAs from Tumor necrosis factor alpha on mesenchymal stem cells and its exosomes.subsequently,to analysis the target gene and its correspondingpathway.Methods:Tumor necrosis factor alpha(TNF-?)used as a stimulator(20ng/ml)on mesenchymal stem cells,which regarded as group TNF-?-cell(TC),another as group Normal Control-cell(NCC)in vice versa;MSCs and their culture supernatant were gathered after 48 hours,respectively.Subsequently,exosomes were isolated from culture supernatants with ExoQuick-TC,which divided into groups TNF-?-exosomes(TE)and Normal Control-exosomes(NCE).Then,RNAs were extracted from every group and picked up their microRNAs;After that microRNAs were measured by High-through Put Sequence and the result were differently analyzed.Finally,the correlation analysis of target gene corresponding differently expressed microRNAs by GO Ontalogy and KEGG Pathway analysis.Result:At the differential expression of microRNA level,high through-put sequencing showed that the TC group had 280 microRNAs,the unique up-regulate expression was miR-146a-5p and the most significantly down-regulate expression of miR-150-5p among 279 microRNAs were included.Identically,there are 180 different expression microRNAs in group TE that miR-146-5p was one of 176 upregulated expressions and miR-203b-5p was one of 4 downregulated expression.Coincidently,bioinformatics analysis showed that IRAK1 was a critical gene of miR-146-5p related to TOLL-like Receptor signaling pathway.Conclusion:In contrast with their control group,there were significantly different expression microRNAs in both MSCs and exosomes;Interestingly,mi R-146a-5p was up-regulate expressed in both comparative groups and its target gene IRAK1 which is playing a crucial part in TOLL-like Receptor signaling pathway according to KEGG Pathway.These investigations indicated a new idea for subsequent inflammation mechanism study. |
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