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

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 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.

Mesothelial cells in tissue repair and fibrosis

“…The prolonged expression of these factors during peritoneal inflammation delays the regression of mesothelial cells back to their epithelial phenotype thereby promoting fibrotic changes in the peritoneum. Other factors recently identified to be associated with MMT include MCP-1 ( Lee et al, 2012 ), ROS ( Liu et al, 2012 ), and the small non-coding regulatory microRNAs miR-589 ( Zhang et al, 2012 ), miR-30a ( Zhou et al, 2013 ), miR-30b ( Liu et al, 2014a ), and miR-200c ( Zhang et al, 2013 ).…”

miR-450a-5p eliminats MGO-induced insulin resistance via targeting CREB

“…In abnormal conditions, the obstacle of MGO clearing pathway will lead to the gradual accumulation of MGO content, which can not only directly cause damage to various cells in the body, but also induce the mass-generation of advanced glycation end products [3]. In previous studies, miRNAs have been proved to be closely related to the content of MGO, such as miR-30b [18], miR-214 [19], miR -9a-3p [20], etc. In this study, the expression of miR-450a-5p showed a downregulated trend in HUVECs with high-glucose status, which was also decreased with the increase of MGO concentration.…”

“…MicroRNAs (miRs) are a class of single-stranded non-coding RNAs with length of approximately [18][19][20][21][22][23] nucleotides, which exert post-transcriptional modification via binding to the 3'untranslated region of target gene, so as to affect the stability and translation of mRNA. It is estimated that miRNAs account for about 1% of the genome of nuclear organisms, and the regulatory range of their target genes covered 30% of all genes [10].…”

microRNA Regulation of Peritoneal Cavity Homeostasis in Peritoneal Dialysis

“…A rat EMT model based on repeated exposure to glucose degradation products (GDPs) during 1-2 weeks, using methylglyoxal (MGO), has been investigated by Liu et al [ 29 ]. Total RNA from the peritoneum of rats subjected to this model was analyzed by miRNA array [ 29 ].…”