Proceedings of The Physiological Society

Europhysiology 2018 (London, UK) (2018) Proc Physiol Soc 41, PCB246

Poster Communications

Analysis of Micro-RNAs Participating the Inhibition of Erythropoietin Synthesis by TNF-alpha via Microarray Technology

M. OZKURT1, R. ÖZYURT1, A. KARADAĞ1, T. Hellwig-Bürgel2, S. Kabadere1, N. erkasap1

1. Eskisehir Osmangazi University, Eskisehir, Turkey. 2. Luebeck Physiology Institute, Luebeck, Germany.

Aim: In chronic inflammatory diseases pro-inflammatory cytokines such as TNF-alpha are high amount in circulation and are associated with anemia in most cases. Experimental studies have shown that TNF-alpha which plays a major role in the inflammation inhibits the synthesis of Erythropoietin (Epo), the main stimulant of hematopoiesis, and the mechanism of this raid has not been announced yet (1). In this study, we aimed to find out which micro-RNAs (miRNA) are involved in inhibition of Epo by TNF-alpha. Material-Methods: Non-cytotoxic TNF-alpha doses were determined by MTT assay. Epo-targeted ELISA and qRT-PCR analyzes were performed to find the optimal dose of TNF-alpha that suppressed Epo in HEPG2 cells. Statistical significance was determined by One-Way Anova and Tukey test for post-hoc. After detecting the TNF-alpha dose as 20 ng/ml, we performed miRNA array (Affymetrix Genechip miRNA 4.0 Array) with untreated (n=6) and treated with 20 ng/ml dose of TNF-alpha HEPG2 cells (n=6). Results: Of the 30,404 miRNAs, 99 human miRNAs showed 2 or more fold changes. According to data base, 8 of them had a potential for binding to Epo mRNA: hsa-miR-4459, hsa-miR-663a, hsa-miR-1908-5p, hsa-miR-4433-3p, hsa-miR-149-3p, hsa-miR-4739, hsa-miR-6805-5p, hsa-miR-122-5p. Conclusion: The detection of miRNAs that play a role in inhibition of Epo by TNF-alpha may describe a physiological mechanism deeper. In the next months, silencing studies of the miRNAs that we will perform, may show new treatment opportunities for the anemia, seen in chronic inflammatory diseases.

Where applicable, experiments conform with Society ethical requirements