<p>lncRNA GAS5/miR-452-5p Reduces Oxidative Stress and Pyroptosis of High-Glucose-Stimulated Renal Tubular Cells</p>

Abstract

BackgroundDiabetic nephropathy (DN) is the leading cause of end-stage renal failure worldwide. lncRNAs are demonstrated to improve the DN by changing the expression of miRNAs. This study was aimed to investigate the effect of lncRNA GAS5/miR-452-5p on the inflammation, oxidative stress and pyroptosis of high-glucose-induced renal tubular cells.MethodsHK-2 cells were induced by HG to simulate DN cells. RT-qPCR analysis confirmed the transfection effects and detected the expression of GAS5, NLRP3, caspase1, IL-1β, pro-caspase1, pro-IL-1β, GSDMD-N and miR-452-5p. Western blot analysis determined the protein expression of NLRP3, caspase1, IL-1β, pro-caspase1, pro-IL-1β and GSDMD-N. The expression of GSDMD-N was also verified by immunofluorescence. The levels of TNF-α, IL-6, MCP-1, ROS, MDA and SOD were measured by commercial assay kits, respectively. Dual-luciferase reporter assay indicated that GAS5 could combine with miR-452-5p.ResultsGAS5 expression was decreased in HG-induced HK-2 cells. GAS5 overexpression could decrease the levels of TNF-α, IL-6, MCP-1, ROS and MDA and increase the levels of SOD. Moreover, GAS5 overexpression suppressed the expression of NLRP3, caspase1, IL-1β and GSDMD-N, and the results of immunofluorescence verified the above results. miR-452-5p interference could cause the same changes as GAS5 overexpression for HG-induced HK-2 cells, and GAS5 inhibition could reverse the effect of miR-452-5p interference.ConclusionGAS5 overexpression inhibited the inflammation, oxidative stress and pyroptosis of HG-induced renal tubular cells by downregulating the expression of miR-452-5p.

Long Non-coding RNA GAS5 Maintains Insulin Secretion by Regulating Multiple miRNAs in INS-1 832/13 Cells

“…Therefore, we supposed that GAS regulated the expression of INSR, PIK3R1 and IRS1 through competitively interacting miR-29a-3p, miR-96-3p, and miR-208a-3p, but we cannot exclude other mechanisms play a role in this regulation, such as epigenetic mechanism (Sun et al, 2017). MiR-452-5p and miR-221-3p are two valid targets of GAS5 in renal tubular cells and mesangial cells, respectively, and both of them are involved in diabetic nephropathy (Ge et al, 2019;Xie et al, 2019). However, the levels of miR-452-5p and miR-221-3p were comparable between high GAS5 group and low GAS5 group.…”

“…LncRNA, as a competitive endogenous RNA (ceRNA), binds to miRNA and acts as a miRNA sponge, thereby reducing the activity of miRNA and indirectly up-regulating the expression of miRNA-related target genes (Goustin et al, 2019). It has been reported that GAS5 targets to miR-452-5p and miR-221-3p in renal tubular cells and mesangial cells, which are involved in diabetic nephropathy (Ge et al, 2019;Xie et al, 2019).…”

New Insights into the Mechanisms of Pyroptosis and Implications for Diabetic Kidney Disease

“…In the pyroptosis pathway, GSDMD is cleaved by inflammatory caspases at an aspartate site within the linking loop, thereby generating the self-inhibited group GSDMD-C-terminal (GSDMD-CT) and the active group GSDMD-N-terminal (GSDMD-NT) [ 32 , 33 ]. Inhibitory binding between GSDMD-NT and GSDMD-CT leads to the inactivity of GSDMD.…”

“…Meanwhile, tubular cell injury has something to do with the role of GSDMD-NT in promoting pyroptosis. After transfecting the HG-induced renal tubular epithelial cells (HK-2 cells) with miRNA-452-5p inhibitor, they detected the marked downregulation of the expressions of NLRP3, caspase-1 and GSDMD-NT [ 32 ]. However, the concrete effect of GSDMD-NT on cell pore formation in tubular cells has yet to be tested.…”

“…Notably, with the research trends of epigenetics, the gene therapy of IL-22 and some types of long-non coding RNA (lncRNA) also perform great effects on inhibiting renal cell pyroptosis and, thus, attenuating kidney fibrosis and DKD progression [ 15 , 18 , 32 , 132 , 133 ].…”