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High-resolution magic angle spinning 1H NMR spectroscopy of metabolic changes in rabbit lens after treatment with dexamethasone combined with UVB exposure

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Abstract

Background

Long-term steroid treatment and UVB exposure are well-known cataractogenic factors. The purpose of this study was to investigate metabolic changes in the rabbit lens after long-term dexamethasone treatment in combination with UVB exposure, using high-resolution magic angle spinning proton nuclear magnetic resonance (HR-MAS 1H NMR) spectroscopy to analyse intact lens tissues.

Methods

Rabbits received topical doses of 0.1% dexamethasone or 0.9% saline (50 μl) four times daily for 36 days. On day 37, the eyes were exposed to UVB radiation (2.05 J/cm2). Twenty-four hours later the animals were killed, and HR-MAS 1H NMR spectra of lens tissues were obtained.

Results

More than 15 major metabolites were assigned in NMR spectra of rabbit lenses. The combined treatment with dexamethasone and UVB induced large reductions in the concentration of reduced glutathione, inositols, taurine and lactate compared with normal lenses. Concurrently, the levels of glucose, sorbitol and sorbitol-3-phosphate were increased. After exposure to UVB radiation only, the most significant finding was a decrease in the concentration of lactate. No lens opacities were detected.

Conclusions

HR-MAS 1H NMR spectroscopy was found to be an efficient tool for analysis of intact lens tissues. High-resolution NMR spectra of intact lens tissue enabled metabolic changes to be quantified. Long-term treatment with dexamethasone combined with UVB exposure induced substantial metabolic changes, dominated by osmolytic regulation processes and loss of glutathione.

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Acknowledgements

Čestmír Čejka is gratefully acknowledged for radiometric measurements. This study was supported by grants from The Research Council of Norway, grant no. 304/03/0419 from the Grant Agency of the Czech Republic and grant AVOZ5008914 from the Academy of Sciences of the Czech Republic. J.K. thanks Bruker BioSpin GmbH, Rheinstetten, Germany, for continuous support in development of probes for making HR-MAS a viable method for metabolic profiling in tissue.

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Authors and Affiliations

  1. Faculty of Medicine, Norwegian University of Science and Technology (NTNU), Trondheim, Norway

    Oddbjørn Sæther, Øystein Risa & Anna Midelfart

  2. Faculty of Natural Sciences and Technology, Norwegian University of Science and Technology (NTNU), Trondheim, Norway

    Oddbjørn Sæther, Øystein Risa & Jostein Krane

  3. MR-Senteret, St Olavs Hospital, 7006, Trondheim, Norway

    Oddbjørn Sæther

  4. Department of Eye Histochemistry and Pharmacology, Institute of Experimental Medicine, Academy of Sciences of the Czech Republic, Prague, Czech Republic

    Jitka Čejková

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  1. Oddbjørn Sæther
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  2. Øystein Risa
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Correspondence to Oddbjørn Sæther.

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Sæther, O., Risa, Ø., Čejková, J. et al. High-resolution magic angle spinning 1H NMR spectroscopy of metabolic changes in rabbit lens after treatment with dexamethasone combined with UVB exposure. Graefe's Arch Clin Exp Ophthalmol 242, 1000–1007 (2004). https://doi.org/10.1007/s00417-004-1030-8

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  • DOI: https://doi.org/10.1007/s00417-004-1030-8

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