Silencing of MALAT1 Reduces Lipopolysaccharide-Induced Glomerular Endothelial Cell Damage via Mediating MiR-126-5p Expression

Abstract

Background: The involvement of long non-coding RNA (lncRNA) metastasis associated lung adenocarcinoma transcript 1 (MALAT1) in inflammation-related kidney damage has been widely demonstrated. However, the mechanisms by which MALAT1 regulate glomerular endothelial cells in sepsis remain undefined, which is the subject in this study. Methods: MALAT1-targeted gene was predicted, and verified through Starbase and dual-luciferase reporter experiment. ShMALAT1 and miR-126-5p inhibitors were transfected into lipopolysaccharide (LPS)-induced human renal glomerular endothelial cells (HRGECs) and murine glomerular endothelial cells (MGECs). The viability, reactive oxygen species (ROS) content, apoptosis and expressions of inflammatory cytokines of HRGECs and MGECs were examined by cell counting kit-8, immunofluorescence assay, flow cytometry and enzyme-linked immunosorbent assay (ELISA), respectively. Expressions of vascular cellular adhesion molecule 1 (VCAM1), inter-cellular adhesion molecule-1 (ICAM-1), monocyte-chemoattractant protein-1 (MCP-1), Nuclear P65 and Cytoplasmic P65 were analyzed by Western blotting. Results: MALAT1 silencing promoted viability and Cytoplasmic P65 expression, but inhibited ROS content, apoptosis, inflammatory cytokine (TNF-α (tumor necrosis factor-alpha) and IL-6 (interleukin 6)) levels, and VCAM1, ICAM-1, MCP-1 and Nuclear P65 expressions in LPS-induced HRGECs and MGECs. MiR-126-5p had a targeted binding relationship with MALAT1 in HRGECs, and MALAT1 silencing increased miR-126-5p expression. MiR-126-5p inhibitor had the opposite effect to MALAT1 silencing, and reversed the effect of MALAT1 silencing on viability, apoptosis and inflammatory cytokines in LPS-induced HRGECs and MGECs. Moreover, miR-126-5p inhibitor increased VCAM1, ICAM-1, MCP-1 and Nuclear P65 expressions, but reduced that of Cytoplasmic P65 in LPS-induced HRGECs and MGECs, and partially offset the effect of shMALAT1 on the above protein expressions. Conclusions: MALAT1 silencing reduces LPS-induced glomerular endothelial cell damage by promoting miR-126-5p expression.