ObjectiveTo explore the regulation mechanism of high-concentration dexamethasone on small non-coding RNA (sncRNA) expressions of bone marrow mesenchymal stem cells (BMSCs). MethodsBMSCs of mice were isolated and cultured in vitro; meanwhile, the negative control group (NC group) and the dexamethasone group (DEX group) were induced by the solutions including 0.1 μmol/L or 10 μmol/L dexamethasone, respectively. The states of BMSCs osteogenic differentiations were analyzed via alizarin red staining. The high throughput small RNA sequencing was conducted, the differential expressions of sncRNAs in both NC and DEX groups were screened, and the bio-informatics was analyzed by employing the Illumina Hiseq 2500 platform. ResultsTo compared with the NC group, the DEX group had a lighter alizarin red staining based on results of alizarin red staining, and yielded significantly lower content of calcium nodule via results of the semi-quantitative detection. There were 25 micro RNAs (miRNAs) with |log2Fold Change| ≥1 of the differential genes in both DEX and NC groups, among which in the DEX group, 6 miRNAs expressed in up-regulation, whereas 19 miRNAs in down-regulated expressions (all P＜0.05). The Gene Ontology (GO) results interpreted that the target genes of up-regulated differential miRNAs were mainly involved in biological processes of protein phosphorylation, muscle and organs developments, and cell adhesion etc., and the target genes of down-regulated differential miRNAs were mainly involved in the biological processes of protein auto-phosphorylation, protein ubiquitination, intracellular estrogen receptor signaling pathway, and cell proliferation etc.. The Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that up-regulated differential miRNAs were mainly enriched in fat synthesis related signaling pathway, Ras or calcium signaling pathway etc., whereas down-regulated differential miRNAs were mainly enriched in Wnt or Hippo signaling pathway etc.. Among those differential genes in both DEX and NC groups, there were 3 PIWI-interacting RNAs (piRNAs) with |log2Fold Change| ≥1, thereinto the DEX group had 1 piRNA in up-regulated expression, whereas 2 piRNAs in down-regulated expressions; furthermore, there were 2 repeat associated small interfering RNAs (rasiRNAs) with |log2Fold Change| ≥1, all expressed in up-regulation in the DEX group; moreover, 8 small nucleolar RNAs (snoRNAs) interpreted in |log2Fold Change| ≥1, of which the DEX group had 6 in up-regulated expressions, whereas 2 in down-regulation; in addition, only one small nuclear RNA (snRNA) expressed in |log2Fold Change| ≥1, which was down-regulated expression in the DEX group (all P＜0.05). ConclusionThe sequencing technology of high throughput small RNA can effectively detect the differential expressions of sncRNAs in BMSCs of mice treated with high-concentration dexamethasone, and the analysis of signaling pathways presented that differential expressed miRNAs may involve in multiple biological processes of fat and osteogenesis metabolism etc..

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