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Comparative lipidomic analysis of inflammatory mediators in the aqueous humor and tear fluid of humans and rabbits



Ocular inflammation is a key pathogenic factor in most blindness-causing visual disorders. It can manifest in the aqueous humor (AH) and tear fluid (TF) as alterations in polyunsaturated fatty acids (PUFAs) and their metabolites, oxylipins, lipid mediators, which are biosynthesized via enzymatic pathways involving lipoxygenase, cyclooxygenase or cytochrome P450 monooxygenase and specifically regulate inflammation and resolution pathways.


This study aimed to establish the baseline patterns of PUFAs and oxylipins in AH and TF by their comprehensive lipidomic identification and profiling in humans in the absence of ocular inflammation and comparatively analyze these compounds in the eye liquids of rabbits, the species often employed in investigative ophthalmology.


Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used for qualitative and quantitative characterization of lipid compounds in the analyzed samples.


A total of 28 lipid compounds were identified, including phospholipid derivatives and PUFAs, as well as 22 oxylipins. Whereas the PUFAs included arachidonic, docosahexaenoic and eicosapentaenoic acids, the oxylipins were derived mainly from arachidonic, linoleic and α-linolenic acids. Remarkably, although the concentration of oxylipins in AH was lower compared to TF, these liquids showed pronounced similarity in their lipid profiles, which additionally exhibited noticeable interspecies concordance.


The revealed correlations confirm the feasibility of rabbit models for investigating pathogenesis and trialing therapies of human eye disorders. The identified metabolite patterns suggest enzymatic mechanisms of oxylipin generation in AH and TF and might be used as a reference in ocular inflammation studies.

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Arachidonic acid




Aqueous humor


α-Linolenic acid




Cytochrome P450 monooxygenase


Dry eye syndrome


Docosahexaenoic acid


Dihydroxyoctadecadienoic acid


Dihydroxyoctadecamonoenoic acid


Eicosapentaenoic acid


Epoxyoctadecadienoic acid


Epoxyoctadecamonoenoic acid


Hydroxydocosahexaenoic acid


Hydroxyeicosapentaenoic acid


Hydroxyeicosatetraenoic acid


Hydroxyoctadecadienoic acid


Hydroxyoctadecatrienoic acid


Hydroperoxyoctadecadienoic acid


Hydroperoxyoctadecatrienoic acid


Linoleic acid






Lyso-platelet-activating factor






Polyunsaturated fatty acids


Tear fluid


Ultra-performance liquid chromatography-tandem mass spectrometry


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This study was supported by the Russian Science Foundation (Project No. 16-15-00255). The authors acknowledge the Preclinical Clinical Study Centre of RUDN University and “RUDN University Program 5-100” for their contribution to the development of the procedure of UPLC-MS/MS analysis.

Author information

DVC, MGS, AAZ, PPP and EYZ conceived and designed research. VIK, EVF, OAK, AMB and ENI performed studies involving human participants. VVT, VEB and IIS conducted experiments with animals. DVC, NVA, AAA, SVG and VVC carried out mass-spectrometry analysis. SVG and VVC contributed new reagents or analytical tools. DVC, MGS, AAZ, PPP and EYZ analyzed data. DVC, NVA and EYZ visualized the results. EYZ wrote the manuscript. All authors read and approved the manuscript.

Correspondence to Dmitry V. Chistyakov or Evgeni Yu. Zernii.

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All participants signed a written declaration of consent approved by the Ethics Committee of Helmholtz National Medical Research Center of Eye Diseases (Moscow, Russia).

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The experimental procedures followed the tenets of The Code of Ethics of the World Medical Association (Declaration of Helsinki). The treatment of the animals was performed according to the 8th edition “Guide for the Care and Use of Laboratory Animals” of the National Research Council and “Statement for the Use of Animals in Ophthalmic and Visual Research” of The Association for Research in Vision and Ophthalmology (ARVO). The study protocols were approved by the Belozersky Institute of Physico-chemical Biology Animal Care and Use Committee (Protocol Number 1/2016).

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Chistyakov, D.V., Azbukina, N.V., Astakhova, A.A. et al. Comparative lipidomic analysis of inflammatory mediators in the aqueous humor and tear fluid of humans and rabbits. Metabolomics 16, 27 (2020). https://doi.org/10.1007/s11306-020-1650-y

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  • Aqueous humor
  • Tear fluid
  • PUFAs
  • Oxylipins
  • COX
  • LOX
  • CYP450