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Einfluss von Ischämie/Hypoxie auf die HIF-1-Aktivität und Expression von hypoxieabhängigen Genen in der Kochlea der neugeborenen Ratte

Influence of ischemia/hypoxia on the HIF-1 activity and expression of hypoxia-dependent genes in the cochlea of the newborn rat

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Zusammenfassung

Hintergrund

Der Transkriptionsfaktor HIF-1 („hypoxia-inducible factor-1“) reguliert unter Ischämie/Hypoxie die Expression von Genen für Glukoseversorgung, Wachstum, Stoffwechsel, Redoxreaktionen und Durchblutung. Im Hinblick auf Tinnitus und Schwerhörigkeit wurden HIF-1-Aktivität und Expression von HIF-1-abhängigen Genen in der Kochlea unter Ischämie/Hypoxie untersucht.

Material/Methoden

HIF-1 wurde in einer Einzelzellkultur der Regionen Corti-Organ (OC), Stria vascularis (SV) und Modiolus (MOD), die mRNA-Expression in der organotypischen Kultur mittels Microarray-Technik (RN U34-Chip, Affymetrix) bestimmt.

Ergebnisse

Ischämie (Hypoxie + Glukosemangel) führt zu einer ähnlichen HIF-1-Aktivierung wie reine Hypoxie mit dem höchsten Anstieg im MOD und OC. Der HIF-1α-mRNA-Gehalt ist in der SV höher als im OC und MOD. Während der Kultivierung werden HIF-1α-mRNA und zahlreiche HIF-1-abhängige Gene wie Gapdh/“Glyceraldehyde-3-phosphate dehydrogenase“, Slc2a1/“Solute carrier family 2 (facilitated glucose transporter), member 1“, Tf/Transferrin und Tfrc/Transferrin Rezeptor in allen 3 Regionen hochreguliert. In SV, MOD und OC erfolgt eine besonders hohe Zunahme der Expression von Hmox1/Hämoxygenase 1, Nos2/induzierbare Stickstoffmonoxid-Synthase und Tfrc. Hypoxie (5 h) führt zu einer Erhöhung der Expression von Igf2/“insulin-like growth factor 2“.

Fazit

Die Daten unterstreichen die Bedeutung radikalbildender Prozesse für die Innenohrschädigung. Die organotypische Kultur geht möglicherweise mit einer Hypoxie einher.

Abstract

Background

Transcription factor HIF-1 (hypoxia-inducible factor-1) regulates the expression of genes which are involved in glucose supply, growth, metabolism, redox reactions and blood supply. Hypoxia and ischemia play an important role in the pathogenesis of tinnitus and hearing loss. Therefore, HIF-1 activity and the expression of HIF-1 dependent genes in the cochlea were examined under ischemic and hypoxic conditions.

Material and methods

For the HIF-1 analysis, single-cell cultures of the organ of Corti (OC), stria vascularis (SV) and modiolus (MOD) were used. mRNA expression was analyzed in the organotypic culture using a microarray technique (RN U34-chip, Affymetrix).

Results

Ischemia (hypoxia without glucose) and pure hypoxia increase the HIF-1 activity identically, with the highest increase found in MOD and OC. The HIF-1 α mRNA levels were found to be higher in SV than in the OC and MOD. During culturing, there is a clear increase in HIF-1 α mRNA and the expression of a number of HIF-1 dependent genes, such as Gapdh/glyceraldehyde-3-phosphate dehydrogenase, Slc2a1/solute carrier family 2 (facilitated glucose transporter), member 1, Tf/transferrin and Tfrc/transferrin receptor, in all three regions. In SV, MOD and OC, increase in the expression of Hmox1/hemoxygenase 1, Nos2/nitric oxide synthase, inducible and Tfrc is particularly high. Hypoxia (5 h) results in an increased expression of Igf2/Insulin-like growth factor 2.

Conclusion

The present data underline the contribution of radical forming processes to the pathogenesis of inner ear diseases. For experimental research, it is important to note that organotypic culture may be coupled with hypoxia.

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Authors and Affiliations

  1. Molekularbiologisches Forschungslabor der HNO-Klinik, Charité — Universitätsmedizin Berlin

    B. Mazurek, C. Rheinländer, F.-U. Fuchs, N. Amarjargal, H. Haupt &  J. Gross

  2. Labor für Funktionelle Genomforschung, Charité — Universitätsmedizin Berlin

    R.-J. Kuban & U. Ungethüm

  3. Fachbereich Chemie/Biochemie, Technische Universität Kaiserslautern

    T. Kietzmann

  4. Molekularbiologisches Forschungslabor der HNO-Klinik, Charité — Universitätsmedizin Berlin, Spandauer Damm 130, Haus 31, 14050, Berlin

    J. Gross

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  1. B. Mazurek
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  7. H. Haupt
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Correspondence to J. Gross.

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Mazurek, B., Rheinländer, C., Fuchs, FU. et al. Einfluss von Ischämie/Hypoxie auf die HIF-1-Aktivität und Expression von hypoxieabhängigen Genen in der Kochlea der neugeborenen Ratte. HNO 54, 689–697 (2006). https://doi.org/10.1007/s00106-005-1371-6

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  • DOI: https://doi.org/10.1007/s00106-005-1371-6

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