Lack of miRNA-17 family mediates high glucose-induced PAR-1 upregulation in glomerular mesangial cells.

Zhuang-Zhuang Tang,Ling-Shan Gou,Xiao-Fei An,Yao-Wu Liu,Pan-Pan Gu

doi:10.1007/s00210-021-02184-1

Abstract

Upregulation of thrombin receptor protease-activated receptor 1 (PAR-1) is verified to contribute to chronic kidney diseases, including diabetic nephropathy; however, the mechanisms are still unclear. In this study, we investigated the effect of PAR-1 on high glucose-induced proliferation of human glomerular mesangial cells (HMCs), and explored the mechanism of PAR-1 upregulation from alteration of microRNAs. We found that high glucose stimulated proliferation of the mesangial cells whereas PAR-1 inhibition with vorapaxar attenuated the cell proliferation. Moreover, high glucose upregulated PAR-1 in mRNA level and protein expression while did not affect the enzymatic activity of thrombin in HMCs after 48h culture. Then high glucose induced PAR-1 elevation was likely due to the alteration of the transcription or post-transcriptional processing. It was found that miR-17 family members including miR-17-5p, -20a-5p, and -93-5p were significantly decreased among the eight detected microRNAs only in high glucose-cultured HMCs, but miR-129-5p, miR-181a-5p, and miR-181b-5p were markedly downregulated in both high glucose-cultured HMCs and equivalent osmotic press control compared with normal glucose culture. So miR-20a was selected to confirm the role of miR-17 family on PAR-1 upregulation, finding that miR-20a-5p overexpression reversed the upregulation of PAR-1 in mRNA and protein levels induced by high glucose in HMCs. In summary, our finding indicated that PAR-1 upregulation mediated proliferation of glomerular mesangial cells induced by high glucose, and deficiency of miR-17 family resulted in PAR-1 upregulation.

Highlights

Diabetic nephropathy (DN) is one of the most common diabetic microvascular complications, characterized by continuous proteinuria, glomerular mesangial expansion, abnormal accumulation of extracellular matrix, and thickening of glomerular basement membrane, eventually developing glomerular sclerosis (Chen et al, 2019)
Whether microRNAs take part in the up-regulation of protease-activated receptor 1 (PAR-1) caused by high glucose in glomerular mesangial cells, there are no reports
HG significantly induced cell proliferation in human glomerular mesangial cells (HMCs) after 48 h culture compared with the NG culture (p < 0.01, Fig. 1A), while co-treatment with a selective inhibitor of PAR-1 Vor inhibited cell proliferation induced by HG in HMCs (p < 0.05, Fig. 1A)

Summary

Introduction

Diabetic nephropathy (DN) is one of the most common diabetic microvascular complications, characterized by continuous proteinuria, glomerular mesangial expansion, abnormal accumulation of extracellular matrix, and thickening of glomerular basement membrane, eventually developing glomerular sclerosis (Chen et al, 2019). Recent studies further demonstrate that PAR-1 deficiency or inhibition protects against DN in streptozotocin-induced diabetic mice (Waasdorp et al, 2016, Waasdorp et al, 2018). Our recent study further demonstrated that PAR-1 up-regulation participated in the early pathological process of DN, and the NLRP3 inflammasome and NF-κB signaling mediated the proinflammatory effects of PAR-1 (Tang et al, 2020). Our study indicated that PAR-1 up-regulation was not associated with thrombin activity in both the renal cortex of diabetic rats and high glucose-cultured glomerular mesangial cells, and discussed the possible reasons (Tang et al, 2020). How high glucose induced PAR-1 up-regulation in renal cortex is still unknown

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Conclusion