Abstract

Type 2 diabetic mellitus (DM2) is associated with accelerated thrombotic complications and is characterized by high levels of plasminogen activator inhibitor-1 (PAI-1). Recent studies show that human platelets have high levels of miR-30c and synthesize considerable active PAI-1. The underlying mechanism of how PAI-1 expression is upregulated in DM2 is poorly understood. We now report that hyperglycaemia-induced repression of miR-30c increases PAI-1 expression and thrombus formation in DM2. Bioinformatic analysis and identification of miRNA targets were assessed using luciferase assays, quantitative real-time PCR and western blots in vitro and in vivo. The changes in miR-30c and PAI-1 levels were identified in platelets from healthy and diabetic individuals. We found that miR-30c directly targeted the 3′ UTR of PAI-1 and negatively regulated its expression. miR-30c was negatively correlated with glucose and HbA1c levels in DM2. In HFD-fed diabetic mice, increasing miR-30c expression by lenti-miR-30c significantly decreased the PAI-1 expression and prolonged the time to occlusion in an arterial thrombosis model. Platelet depletion/reinfusion experiments generating mice with selective ablation of PAI-1 demonstrate a major contribution by platelet-derived PAI-1 in the treatment of lenti-miR-30c to thrombus formation. These results provide important implications regarding the regulation of fibrinolysis by platelet miRNA under diabetic mellitus.

Highlights

  • This study, using purified human platelets, we demonstrated the existence of a miR30c-targeted PAI-1 pathway in platelets of the cardiovascular system

  • We confirmed that miR-30c (MIMAT0000244) and PAI-1 (NM_000602.3) mRNA were found in the small/total RNA libraries of leukocyte-depleted platelets (LDP) by PCR assay from healthy individuals

  • LDP isolated from healthy individuals and patients with pre-DM, NCDM and DM-CHD were measured as described above (Fig. 1D), indicating the marked and successful depletion of leukocytes from the starting platelet-rich plasma (PRP)

Read more

Summary

Introduction

This study, using purified human platelets, we demonstrated the existence of a miR30c-targeted PAI-1 pathway in platelets of the cardiovascular system. The gene expression levels of miR-30c and PAI-1 in LDPs were measured by qRT-PCR, and the PAI-1 total antigen was determined by ELISA as described above. The PAI-1 mRNA expression levels were up-regulated and significantly greater in the DM-CHD subjects compared with other groups (Fig. 2C,D).

Results
Conclusion
Full Text
Published version (Free)

Talk to us

Join us for a 30 min session where you can share your feedback and ask us any queries you have

Schedule a call