Protective Effects of Microrna-22 Against Endothelial Cell Injury by Targeting NLRP3 Through Suppression of the Inflammasome Signaling Pathway in a Rat Model of Coronary Heart Disease

Cell Physiol Biochem. 2017;43(4):1346-1358. doi: 10.1159/000481846. Epub 2017 Oct 9.

Abstract

Background/aims: This study aimed to identify the role of microRNA-22 (miR-22) in endothelial cell (EC) injury in coronary heart disease (CHD) by targeting NLRP3 through the inflammasome signaling pathway.

Methods: A total of 24 healthy male Sprague-Dawley (SD) rats were divided into normal and atherosclerosis groups. The atherosclerosis rats were assigned into blank, negative control (NC), miR-22 mimic, miR-22 inhibitor and miR-22 inhibitor + siNLRP3 groups. A luciferase reporter gene assay was used to detect the relationship between miR-22 and NLRP3. MiR-22 expression as well as NLRP3 and caspase-1 mRNA and protein expression were measured using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. The activity and apoptosis of coronary arterial endothelial cells (CAECs) were determined by MTT and Hoechst 33258. CAEC lumen formation was detected by a lumen formation assay. An enzyme-linked immunosorbent assay (ELISA) was used to detect IL-1β, IL-6, IL-10 and IL-18 levels.

Results: The results indicated that the atherosclerosis group significantly decreased miR-22 expression but increased NLRP3 and caspase-1 mRNA and protein expression. The cell survival rate was significantly increased in the miR-22 mimic group and significantly reduced in the miR-22 inhibitor group. The miR-22 mimic group displayed a lower apoptosis rate and more cells with obvious lumen walls and numerous tubular structures, while cells in the miR-22 inhibitor group were unable to form lumen walls and had a scattered distribution compared to the blank group. The ELISA showed that IL-1β, IL-6 and IL-18 levels were markedly decreased, while IL-10 was clearly increased in the miR-22 mimic group. In contrast, in the miR-22 inhibitor group, IL-1β, IL-6 and IL-18 levels were significantly increased, and IL-10 levels were decreased.

Conclusion: Our findings indicated that miR-22 could lower the levels of pro-inflammatory cytokines by inhibiting the NLRP3 inflammasome pathway, which suppresses CAEC apoptosis and protects CAECs in rats with CHD.

Keywords: Apoptosis; Coronary heart disease; Endothelial cells; Inflammasome; Microrna-22; NLRP3; Signaling pathway.

Publication types

  • Retracted Publication

MeSH terms

  • Animals
  • Apoptosis*
  • Coronary Disease / genetics*
  • Coronary Disease / immunology
  • Coronary Disease / pathology
  • Endothelial Cells / immunology
  • Endothelial Cells / metabolism
  • Endothelial Cells / pathology*
  • Gene Expression Regulation
  • Inflammasomes / immunology*
  • Interleukin-18 / analysis
  • Interleukin-18 / immunology
  • Interleukin-1beta / analysis
  • Interleukin-1beta / immunology
  • Interleukin-6 / analysis
  • Interleukin-6 / immunology
  • Male
  • MicroRNAs / genetics*
  • MicroRNAs / immunology
  • NLR Family, Pyrin Domain-Containing 3 Protein / genetics*
  • NLR Family, Pyrin Domain-Containing 3 Protein / immunology
  • Rats
  • Rats, Sprague-Dawley
  • Signal Transduction

Substances

  • Inflammasomes
  • Interleukin-18
  • Interleukin-1beta
  • Interleukin-6
  • MIRN22 microRNA, rat
  • MicroRNAs
  • NLR Family, Pyrin Domain-Containing 3 Protein
  • Nlrp3 protein, rat