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MiR-30b is involved in methylglyoxal-induced epithelial-mesenchymal transition of peritoneal mesothelial cells in rats

Cellular & Molecular Biology Letters volume 19, pages 315–329 (2014)Cite this article

Abstract

Epithelial-mesenchymal transition (EMT) of peritoneal mesothelial cells (PMC) is a major contributor to the pathogenesis of peritoneal fibrosis. EMT is at least in part caused by repeated exposure to glucose degradation products (GDPs), such as methylglyoxal (MGO). MiRNA contributes greatly to the EMT of PMCs. In this study, we tried to profile whether differences exist between the peritoneal membrane (PM) miRNA expression seen in control rats and that seen in rats injected intraperitoneally with MGO. We assessed whether miR-30b has a possible role in MGO-induced EMT of PMCs in rats. Comparative miRNA expression array and real-time PCR analyses were conducted for the control group at the start of the experiment and for the MGO group after 1 and 2 weeks. During the second week, the MGO rats were treated with: a chemically modified antisense RNA oligonucleotide (ASO) complementary to the mature miR-30b (ASO group); an miR-30b mismatch control sequence (MIS group); or a citrate buffer (EMT group). Bioinformatic analyses indicated that the 3′ untranslated region (3′-UTR) of bone morphogenetic protein 7 (BMP7) mRNA did contain a putative binding site for miR-30b. We also tried to investigate whether miR-30b targeted BMP7 in vitro by transfection. Of the upregulated miRNAs, miR-30b expression demonstrated the greatest increase. The administration of miR-30b ASO for two weeks significantly reduced α-SMA excretion and upregulated E-cadherin and BMP-7 expression. Our in vitro study showed that miR-30b directly targeted and inhibited BMP7 by binding to its 3’-UTR. Our results revealed that miR-30b is involved in MGO-induced EMT of PMCs in rats.

Abbreviations

3′-UTR:

3′ untranslated region

α-SMA:

α-smooth muscle actin

ASO:

antisense RNA oligonucleotide

BMP7:

bone morphogenetic protein 7

EDTA:

ethylene diamine tetraacetic acid

EMT:

epithelial-mesenchymal transition

ESRD:

endstage renal disease

GDPs:

glucose degradation products

HRP:

horseradish peroxidase

MGO:

methylglyoxal

PD:

peritoneal dialysis

PDFs:

peritoneal dialysis fluids

PM:

peritoneal membrane

PMC:

peritoneal mesothelial cells

RPMCs:

rat peritoneal mesothelial cells

RRT:

renal replacement therapy

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Authors and Affiliations

  1. Department of Emergency Medicine, People’s Hospital of Lishui, Zhejiang Province, 15 Dazhong Street, Liandu District, Lishui, 323000, Zhejiang Province, P.R. China

    Hong Liu

  2. Department of Critical Care Medicine, People’s Hospital of Lishui, Zhejiang Province, 15 Dazhong Street, Liandu District, Lishui, 323000, Zhejiang Province, P.R. China

    Ning Zhang

  3. Department of Neurosurgery, People’s Hospital of Lishui, Zhejiang Province, 15 Dazhong Street, Liandu District, Lishui, 323000, Zhejiang Province, P.R. China

    Da Tian

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  1. Hong Liu
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Correspondence to Da Tian.

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Liu, H., Zhang, N. & Tian, D. MiR-30b is involved in methylglyoxal-induced epithelial-mesenchymal transition of peritoneal mesothelial cells in rats. Cell Mol Biol Lett 19, 315–329 (2014). https://doi.org/10.2478/s11658-014-0199-z

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Keywords

Cellular & Molecular Biology Letters

ISSN: 1689-1392

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