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Quantitative metabolomic analysis of the human cornea and aqueous humor

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Abstract

Introduction

Cornea is the outermost part of the eye supplied mostly by aqueous humor (AH). Therefore, the comparison of the metabolomic compositions of AH and cornea may help to determine which compounds are produced inside the cornea, and which penetrate into cornea from AH for intra-corneal consumption. Keratoconus (KC) is the most common form of the cornea dystrophy, and the analysis of KC corneas can unravel the metabolomic changes occurring in AH and cornea of KC patients.

Objectives

The work is aimed at the determination of concentrations of a wide range of metabolites in the human cornea and AH, the comparison of the metabolomic profiles of cornea and AH, and the comparison of the metabolomic compositions of samples taken from KC patients and normal donors (post-mortem).

Methods

The quantitative metabolomic profiling was carried out with the use of two independent methods—high-frequency 1H NMR spectroscopy and HPLC with high-resolution ESI-MS detection.

Results

The concentrations of 71 most abundant metabolites in cornea and AH from keratoconus patients and from human cadavers have been measured. It is found that the concentrations of purines and organic acids in cornea are significantly higher than in AH. The KC corneas are characterized by the enhanced levels of acetate and citrate, and also by low values of GSH/GSSG ratios.

Conclusion

A significant difference in the metabolomic compositions of the human AH and cornea has been revealed. The concentrations of glucose and some amino acids in cornea are significantly lower than in AH, indicating their fast consumption inside the cornea. The high levels of organic acids, purines and GSH in cornea should be attributed to their production in the cornea. The enhanced levels of acetate and citrate as well as the low values of GSH/GSSG ratios in KC corneas are the indicators of the oxidative stress.

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Acknowledgements

The work was supported by the SB RAS (Program “Integration and development” II.2П/V.44-18, project 0333-2016-0004) in NMR measurements, by RFBR (projects 17-03-00656) in LC-MS measurements, and by FASO Russia (project 0333-2016-0001) in sample preparation.

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Authors and Affiliations

  1. International Tomography Center SB RAS, Institutskaya 3a, Novosibirsk, 630090, Russia

    Olga A. Snytnikova, Lyudmila V. Yanshole, Vadim V. Yanshole & Yuri P. Tsentalovich

  2. Novosibirsk State University, Pirogova 2, Novosibirsk, 630090, Russia

    Olga A. Snytnikova, Lyudmila V. Yanshole, Vadim V. Yanshole & Yuri P. Tsentalovich

  3. Multidisciplinary Science and Technology Complex “Eye Microsurgery”, Kolchidskaya 10, Novosibirsk, 630096, Russia

    Igor A. Iskakov & Valery V. Chernykh

  4. Novosibirsk Regional Clinical Bureau of Forensic Medical Examination, Nemirovicha-Danchenko 134, Novosibirsk, 630087, Russia

    Denis A. Stepakov & Vladimir P. Novoselov

Authors
  1. Olga A. Snytnikova
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  2. Lyudmila V. Yanshole
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  3. Igor A. Iskakov
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  4. Vadim V. Yanshole
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  5. Valery V. Chernykh
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  6. Denis A. Stepakov
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  7. Vladimir P. Novoselov
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  8. Yuri P. Tsentalovich
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Corresponding author

Correspondence to Yuri P. Tsentalovich.

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The study was conducted in accordance with the Declaration of Helsinki (2013) of the World Medical Association, and with the ethical approval from International Tomography Center.

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Snytnikova, O.A., Yanshole, L.V., Iskakov, I.A. et al. Quantitative metabolomic analysis of the human cornea and aqueous humor. Metabolomics 13, 152 (2017). https://doi.org/10.1007/s11306-017-1281-0

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