MiR-19a and miR-20a and Tissue Factor Expression in Activated Human Peripheral Blood Mononuclear Cells.

Cristina Balia,Fulvio Basolo,Mirella Giordano,Alessandro Celi,Gabriella Fontanini,Roberto Pedrinelli,Valentina Scalise,Tommaso Neri

doi:10.1155/2017/1076397

Abstract

Background and Aims To investigate the behaviour of miR-19a and miR-20a, two microRNAs involved in posttranscriptional modulation of TF expression in peripheral blood mononuclear cells (PBMCs) exposed to high glucose (HG) and lipopolysaccharide (LPS), and to evaluate the involvement of angiotensin II in that process. Methods TF Procoagulant Activity (PCA, one-stage clotting assay), antigen (Ag, ELISA), and miR-19a and miR-20a levels (specific TaqMan® MicroRNA Assays) were evaluated in PBMCs exposed to high glucose (HG, 50 mM), LPS (100 ng/mL), and Olmesartan (OLM, 10−6 M), an angiotensin II type 1 receptor antagonist. Results HG increased TF expression and decreased both miRs as compared to control glucose conditions (11.1 mM). In HG-activated PBMCs, LPS stimulated TF expression and downregulated miR-20a, an effect reverted by OLM (10−6 M); miR-19a expression was unchanged by LPS in both CG and HG conditions. Conclusions miR-19a and miR-20a are inhibited by inflammatory stimuli active on TF expression and their response differs by the stimulus under investigation; angiotensin II may participate in that mechanism.

Highlights

MicroRNAs are small, ∼22-nucleotide noncoding RNAs that inhibit transcriptional gene expression by interacting with sites of complementarity in the 3󸀠 untranslated regions (3-UTR) of target mRNAs [e.g., [1]]
Posttranscriptional gene modulation by miRs involves several genes including Tissue Factor (TF) [2], the principal initiator of the clotting cascade, and a major regulator of haemostasis and thrombosis [3] expressed by circulating monocytes exposed to proinflammatory stimuli such as lipopolysaccharide (LPS, endotoxin) [3] and high glucose (HG) [4]
That information, obtained in a very specific context, raises the issue of the behaviour of those two noncoding RNAs in response to stimuli active on TF expression in peripheral blood mononuclear cells (PBMCs) harvested from normal subjects activated by HG and LPS and whether ATII is involved in that relationship, an issue that has never been addressed insofar

Summary

Introduction

MicroRNAs (miRs) are small, ∼22-nucleotide noncoding RNAs that inhibit transcriptional gene expression by interacting with sites of complementarity in the 3󸀠 untranslated regions (3-UTR) of target mRNAs [e.g., [1]]. That information, obtained in a very specific context, raises the issue of the behaviour of those two noncoding RNAs in response to stimuli active on TF expression in peripheral blood mononuclear cells (PBMCs) harvested from normal subjects activated by HG and LPS and whether ATII is involved in that relationship, an issue that has never been addressed insofar. To investigate the behaviour of miR-19a and miR-20a, two microRNAs involved in posttranscriptional modulation of TF expression in peripheral blood mononuclear cells (PBMCs) exposed to high glucose (HG) and lipopolysaccharide (LPS), and to evaluate the involvement of angiotensin II in that process. TF Procoagulant Activity (PCA, one-stage clotting assay), antigen (Ag, ELISA), and miR-19a and miR-20a levels (specific TaqMan MicroRNA Assays) were evaluated in PBMCs exposed to high glucose (HG, 50 mM), LPS (100 ng/mL), and Olmesartan (OLM, 10−6 M), an angiotensin II type 1 receptor antagonist. Conclusions. miR-19a and miR-20a are inhibited by inflammatory stimuli active on TF expression and their response differs by the stimulus under investigation; angiotensin II may participate in that mechanism

Methods

Results

Discussion

Conclusion