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Modulation of miRNAs by vitamin C in H2O2?exposed human umbilical vein endothelial cells.

ABSTRACT: Vitamin C plays a protective role in oxidative damage by blocking the effects of free radicals. The present study investigated the mechanisms through which vitamin C partly mediates anti?apoptotic and antioxidant functions via the regulation of microRNAs (miRNAs or miRs). For this purpose, a global miRNA expression analysis on human umbilical vein endothelial cells (HUVECs) treated with vitamin C was conducted using microarrays containing human precursor and mature miRNA probes. The results revealed that there were 42 identical miRNAs among the differentially expressed miRNAs in the HUVEC group and H2O2 + vitamin C?treated HUVEC group compared to the H2O2?exposed HUVEC group, including 41 upregulated miRNAs and 1 down?regulated miRNA. Using bioinformatics analysis, differentially expressed miRNAs were investigated to identify novel target mRNAs and signaling pathways. Pathway enrichment analyses revealed that apoptosis, the mitogen?activated protein kinase (MAPK) signaling pathway, phosphoinositide 3?kinase (PI3K)/Akt signaling pathway and oxidative phosphorylation were significantly enriched. The results from western blot analysis demonstrated that the interleukin (IL)10, matrix metalloproteinase (MMP)2, cAMP?response element binding protein (CREB) and p?CREB protein expression levels in HUVECs transfected with hsa?miR?3928?5p and induced by H2O2 were significantly downregulated; the MAPK9, caspase?3 (CASP3) and p?CASP3 protein expression levels in HUVECs transfected with hsa?miR?323a?5p and induced by H2O2 were significantly downregulated. The present study therefore demonstrates that vitamin C partly exerts protective effects on HUVECs through the regulation of miRNA/mRNA axis expression.

SUBMITTER: Wu J

PROVIDER: S-EPMC7595655 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature

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Modulation of miRNAs by vitamin C in H2O2‑exposed human umbilical vein endothelial cells.

Wu Jiandi J Liang Jingjing J Li Meijun M Lin Mingzhuo M Mai Linlin L Huang Xiaohui X Liang Jianqiu J Hu Yunzhao Y Huang Yuli Y

International journal of molecular medicine 20201013 6

Vitamin C plays a protective role in oxidative damage by blocking the effects of free radicals. The present study investigated the mechanisms through which vitamin C partly mediates anti‑apoptotic and antioxidant functions via the regulation of microRNAs (miRNAs or miRs). For this purpose, a global miRNA expression analysis on human umbilical vein endothelial cells (HUVECs) treated with vitamin C was conducted using microarrays containing human precursor and mature miRNA probes. The results reve ...[more]

PMID: 33125125

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Project description:BACKGROUND: In diabetes chronic hyperinsulinemia contributes to the instability of the atherosclerotic plaque and stimulates cellular proliferation through the activation of the MAP kinases, which in turn regulate cellular proliferation. However, it is not known whether insulin itself could increase the transcription of specific genes for cellular proliferation in the endothelium. Hence, the characterization of transcriptional modifications in endothelium is an important step for a better understanding of the mechanism of insulin action and the relationship between endothelial cell dysfunction and insulin resistance. METHODOLOGY AND PRINCIPAL FINDINGS: The transcriptional response of endothelial cells in the 440 minutes following insulin stimulation was monitored using microarrays and compared to a control condition. About 1700 genes were selected as differentially expressed based on their treated minus control profile, thus allowing the detection of even small but systematic changes in gene expression. Genes were clustered in 7 groups according to their time expression profile and classified into 15 functional categories that can support the biological effects of insulin, based on Gene Ontology enrichment analysis. In terms of endothelial function, the most prominent processes affected were NADH dehydrogenase activity, N-terminal myristoylation domain binding, nitric-oxide synthase regulator activity and growth factor binding. Pathway-based enrichment analysis revealed "Electron Transport Chain" significantly enriched. Results were validated on genes belonging to "Electron Transport Chain" pathway, using quantitative RT-PCR. CONCLUSIONS: As far as we know, this is the first systematic study in the literature monitoring transcriptional response to insulin in endothelial cells, in a time series microarray experiment. Since chronic hyperinsulinemia contributes to the instability of the atherosclerotic plaque and stimulates cellular proliferation, some of the genes identified in the present work are potential novel candidates in diabetes complications related to endothelial dysfunction.

Dataset Information

Modulation of miRNAs by vitamin C in H2O2?exposed human umbilical vein endothelial cells.

Publications

Modulation of miRNAs by vitamin C in H2O2‑exposed human umbilical vein endothelial cells.

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