Hyperglycaemia suppresses microrna expression in platelets to increase P2RY12 and SELP levels in type 2 diabetes mellitus

Zsolt Fejes, Szilárd Póliska, Zsolt Czimmerer, Miklós Káplár, András Penyige, Gabriella Gál Szabó, Ildikó Beke Debreceni, Satya P. Kunapuli, J. Kappelmayer, B. Nagy

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Megakaryocyte (MK)-derived miRNAs have been detected in platelets. Here, we analysed the expression of platelet and circulating miR-223, miR-26b, miR-126 and miR-140 that might be altered with their target mRNAs in type 2 diabetes mellitus (DM2). MiRNAs were isolated from leukocyte-depleted platelets and plasma samples obtained from 28 obese DM2, 19 non-DM obese and 23 healthy individuals. The effect of hyperglycaemia on miRNAs was also evaluated in MKs using MEG-01 and K562 cells under hyperglycaemic conditions after 8 hours up to four weeks. Quantitation of mature miRNA, pre-miRNAs and target mRNA levels (P2RY12 and SELP) were measured by RT-qPCR. To prove the association of miR-26b and miR-140 with SELP (P-selectin) mRNA level, overexpression or inhibition of these miRNAs in MEG-01 MKs was performed using mimics or anti-miRNAs, respectively. The contribution of calpain substrate Dicer to modulation of miRNAs was studied by calpain inhibition. Platelet activation was evaluated via surface P-selectin by flow cytometry. Mature and pre-forms of investigated miRNAs were significantly reduced in DM2, and platelet P2RY12 and SELP mRNA levels were elevated by two-fold at increased platelet activation compared to controls. Significantly blunted miRNA expressions were observed by hyperglycaemia in MEG-01 and K562-MK cells versus baseline values, while the manipulation of miR-26b and miR-140 expression affected SELP mRNA level. Calpeptin pretreatment restored miRNA levels in hyperglycaemic MKs. Overall, miR-223, miR-26b, miR-126 and miR-140 are expressed at a lower level in platelets and MKs in DM2 causing upregulation of P2RY12 and SELP mRNAs that may contribute to adverse platelet function.

Original languageEnglish
Pages (from-to)529-542
Number of pages14
JournalThrombosis and Haemostasis
Volume117
Issue number3
DOIs
Publication statusPublished - 2017

Keywords

  • Diabetes mellitus
  • Hyperglycaemia
  • Megakaryocyte
  • MicroRNA
  • Platelet activation

ASJC Scopus subject areas

  • Hematology

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    Fejes, Z., Póliska, S., Czimmerer, Z., Káplár, M., Penyige, A., Gál Szabó, G., Beke Debreceni, I., Kunapuli, S. P., Kappelmayer, J., & Nagy, B. (2017). Hyperglycaemia suppresses microrna expression in platelets to increase P2RY12 and SELP levels in type 2 diabetes mellitus. Thrombosis and Haemostasis, 117(3), 529-542. https://doi.org/10.1160/TH16-04-0322