MicroRNA and mRNA analysis of angiotensin II‑induced renal artery endothelial cell dysfunction

Abstract

Continuous activation of angiotensin II (Ang II) induces renal vascular endothelial dysfunction, inflammation and oxidative stress, all of which may contribute to renal damage. MicroRNAs (miRs/miRNAs) play a crucial regulatory role in the pathogenesis of hypertensive nephropathy (HN). The present study aimed to assess the differential expression profiles of potential candidate genes involved in Ang II-induced rat renal artery endothelial cell (RRAEC) dysfunction and explore their possible functions. In the present study, the changes in energy metabolism and autophagy function in RRAECs were evaluated using the Seahorse XF Glycolysis Stress Test and dansylcadaverine/transmission electron microscopy following exposure to Ang II. Subsequently, mRNA-miRNA sequencing experiments were performed to determine the differential expression profiles of mRNAs and miRNAs. Integrated bioinformatics analysis was applied to further explore the molecular mechanisms of Ang II on endothelial injury induced by Ang II. The present data supported the notion that Ang II upregulated glycolysis levels and promoted autophagy activation in RRAECs. The sequencing data demonstrated that 443 mRNAs and 58 miRNAs were differentially expressed (DE) in response to Ang II exposure, where 66 mRNAs and 55 miRNAs were upregulated, while 377 mRNAs and 3 miRNAs were downregulated (fold change >1.5 or <0.67; P<0.05). Functional analysis indicated that DE mRNA and DE miRNA target genes were mainly associated with cell metabolism (metabolic pathways), differentiation (Th1 and Th2 cell differentiation), autophagy (autophagy-animal and autophagy-other) and repair (RNA-repair). To the best of the authors' knowledge, this is the first report on mRNA-miRNA integrated profiles of Ang II-induced RRAECs. The present results provided evidence suggesting that the miRNA-mediated effect on the 'mTOR signaling pathway' might play a role in Ang II-induced RRAEC injury by driving glycolysis and autophagy activation. Targeting miRNAs and their associated pathways may provide valuable insight into the clinical management of HN and may improve patient outcome.

Astragalus membranaceus and Salvia miltiorrhiza Ameliorate Hypertensive Renal Damage through lncRNA-mRNA Coexpression Network

“…Further analysis of 13 coexpressed DE-mRNAs indicated that PPAR signaling pathway was highly enriched in KEGG pathways, and negative regulation of blood pressure and fatty acid beta-oxidation was highly enriched in GO pathways. Our previous studies in vitro found that PPAR signaling pathway and renin-angiotensin system were highly involved in Ang II induced injury of renal arterial endothelial cells (RRAECs) by integrative analysis of miRNA-mRNA sequencing [18]. This study in vivo found that PPAR signaling pathway was also closely involved in the regulation of AS on BP and renal damage in SHRs.…”

Mechanisms of inflammation modulation by different immune cells in hypertensive nephropathy

“…Overproduction of Ang II in the kidney directly constricts vascular endothelial cells (ECs), causing changes in diastolic and contractile substances that are involved in hypertension and kidney injury. In hypertensive rats, the miRNA-mediated mTOR signaling pathway may play a role in Ang II-induced renal artery endothelial cell injury by driving glycolysis and activating autophagy ( 32 ).…”

Inhibition of tumor growth in osteosarcoma cell culture with mir162a miicroRNA

“…Препараты клеток для оценки аутофагии получали таким же способом как для оценки апоптоза, некроза, жизнеспособности . Для выявления клеток с индуцированной аутофагией был использован набор Autophagy Assay Kit (Merck, Germany), содержащий MAK-138 согласно протоколу производителя с 30-минутной инкубацией (MAK138 Autophagy Assay Kit Technical bulletin, Sigma-Aldrich, Merck, Germany, last updated 2020) [9]. С помощью микроскопа MCX 300 Orchid HBO (Micros, Austria) в 10 полях зрения подсчитывали количество клеток с индуцированной аутофагией в канале Violet (диапазон 340/425 нм) и общее количество клеток в проходящем свете.…”