Differentially expressed lnc - NOS2P3-miR-939-5p  axis in chronic heart failure inhibits myocardial and endothelial cells  apoptosis via iNOS/TNFα pathway

Abstract Background: Inflammatory cytokines induced cells apoptosis is important for initiation and progression of chronic heart failure (CHF). Long noncoding RNAs (lncRNAs) and microRNAs (miRNAs) were critical in this pathogenesis. However, their roles in inflammation and apoptosis of CHF remain unclear. Methods: A total of 75 CHF and 36 non-CHF control patients were collected and the level of miR-939-5p was detected. Bioinformatics analysis and luciferase reporter assay was used to predict and verify pairs of lncRNA-miRNA-mRNA. RT-qPCR was used to evaluate the expression of lncRNA, miRNAs and mRNAs. CCK8 and flow cytometry were applied to determine cells vitality and apoptosis. Western blot was used for protein level analysis. Results : CHF patients had elevated serum miR-939-5p, with greater increase in NYHA I-II patients than in NYHA III-IV. Moreover, miR-939-5p was positively correlated with BNP in NYHA III-IV patients, while not in NYHA I-II, suggesting that miR-939-5p might be an additional supplement for diagnosis of heart failure. Further study showed miR-939-5p mimic promoted proliferation and inhibited inflammatory cytokines induced apoptosis of HUVECs and H9C2, while inhibition of endogenous miR-939-5p by antagomir produced the opposite effects. INOS and TNFα were identified and confirmed as target genes of miR-939-5p. Moreover, we identified lncRNA-NOS2P3 as a sponge RNA to inhibit miR-939-5p expression, regulate the expression of iNOS/TNFα, and control inflammation induced myocardial and endothelial cells apoptosis. Conclusions : Together, CHF patients exhibited elevated serum miR-939-5p especially in NYHA I-II grades. Lnc-NOS2P3-miR-939-5p-iNOS/TNFα pathway played an important role in regulating inflammatory cytokines induced cells apoptosis and provided a promising strategy for diagnosis and treatment of CHF.