Methylglyoxal-induced miR-223 suppresses rat vascular KATP channel activity by downregulating Kir6.1 mRNA in carbonyl stress

Vascular Pharmacology
Xin JinShan-Shan Li

Abstract

The vascular ATP-sensitive K+ (KATP) channel composed of Kir6.1 and SUR2B subunits regulates cellular activity by coupling intermediary metabolism to membrane excitability. Our previous studies have shown that both Kir6.1 and SUB2B are post-transcriptionally downregulated by methylglyoxal (MGO) which is a reactive carbonyl specie and can cause disruption of vascular tone regulation under diabetic conditions. We have shown that the SUB2B downregulation is mediated by the microRNA (miR) miR-9a, while the mechanism underlying Kir6.1 inhibition is still unclear. Studying the microRNA databases, we found that miR-223 has sequence similarities to the 3' untranslated sequence (3'UTR) of Kir6.1 mRNA suggesting their potential interactions. Therefore, we explored the role of miR-233 in KATP channel regulation by up/down-regulation of miR-223 in smooth muscle cells (SMCs) and mesenteric arterials. Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) analysis showed augmentation of miR-223 expression in the cultured SMCs after 300 μM MGO exposure by 5-6 folds. miR-223 overexpression down-regulated Kir6.1 mRNA levels by ~2.6 times while miR-223 knockdown diminished the effect of 300 μM MGO by ~50% in the SMCs. Luciferase ...Continue Reading

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