Abstract
ObjectiveTo investigate the role of microRNA-155-5p on apoptosis and inflammatory response in human renal glomerular endothelial cells (HRGEC) cultured with high glucose.MethodsThe primary HRGEC were mainly studied, light microscopy was used to detect changes in cell morphology. Quantitative Real Time-Polymerase Chain Reaction, Western Blot, immunofluorescence were aimed to observe the mRNA and protein expression levels of target gene ETS-1, downstream factors VCAM-1, MCP-1 and cleaved caspase-3 in each group after high glucose treatment as well as transfection with miR-155 mimics or inhibitor.ResultsThe expression of inflammatory factors and apoptosis of HRGEC cells increased under high glucose treatment. Compared with normal-glucose treatment, the expression of microRNA-155 markedly increased in HRGECs treated with high-glucose, as well as the mRNA and protein levels of ETS-1, VCAM-1, MCP-1 and cleaved caspase-3. Overexpression of microRNA-155 remarkably downregulated mRNA and protein levels of ETS-1, VCAM-1, MCP-1 and cleaved caspase-3, whereas miRNA-155 knockdown upregulated their levels. In addition, HRGEC cells were transfected with miR-155 mimics and ETS-1 siRNA with high glucose stimulation. The expression of ETS-1 was positively correlated with the expression of downstream factors VCAM-1 and MCP-1. These results suggest that ETS-1 can mediate endothelial cell inflammation by regulating VCAM-1 and MCP-1.ConclusionMiR-155 can negatively regulate the expression of target gene ETS-1 and its downstream factors VCAM-1, MCP-1 and cleaved caspase-3, thus mediating the inflammatory response and apoptosis of HRGEC.
Highlights
Diabetic nephropathy (DN) is a specific complication of long-term poor blood glucose control, which may lead to progressive renal function damage and cardiovascular risk [1, 2]
It was found that cells in the normal glucose (NG), high mannitol (HM) and high glucose (HG) groups had good growth state, full morphology and tight intercellular connections
Compared with HM, HG and transfection control group, a large number of intact and adherent round bubble cells were found in miR-155 inhibitor group, but no such cells were found in HM and HG group, and few cells in miR-155 mimic group
Summary
Diabetic nephropathy (DN) is a specific complication of long-term poor blood glucose control, which may lead to progressive renal function damage and cardiovascular risk [1, 2]. Henan, China 3 Department of Nephrology, Lanzhou University Second. No 82, Cuiyingmen, Lanzhou 730030, Gansu, China which brings heavy burden to our country and society. Current studies indicate that the DN is caused by a combination of multiple factors, such as elevated blood glucose, changes in renal hemodynamics caused by hypertension, ethnic risk, genetic and metabolic interactions, etc. The molecular level of DNA methylation, chromatin histone modification, new transcripts and functional non-coding RNAs, for example, microRNA and long non-coding RNA have been rapidly studied in the fields of glomerular immune inflammatory response, epithelial mesenchymal transformation, cell apoptosis, mitochondrial damage, podocyte endothelial cell interaction, etc. This study selected the primary human glomerular endothelial cells as the research object, because it is located in the
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