“…The group of Song et al have studied the metabolic changes in rabbit AH after glucocorticosteroids administration [15] and the hypotensive effects of glycyrrhizin on a rabbit model of ocular hypertension induced by triamcinolone acetonide [16]. 1 H NMR has also been used to study metabolic alterations in AH in the rat glaucoma model induced by intracameral sodium hyaluronate injections [17] and to evaluate the effect of UV-A and UV-B irradiation on the metabolic profile of aqueous humor in rabbits [18]. Currently there is only one metabolomics study on human aqueous humor.…”
Aqueous humor (AH) is a transparent fluid which fills the anterior and posterior chambers of the eye. It supplies nutrients and removes metabolic waste from avascular tissues in the eye. Proper homeostasis of AH is required to maintain adequate intraocular pressure as well as optical and refractive properties of the eye. Application of metabolomics to study human AH may improve knowledge about the molecular mechanisms of eye diseases. Until now, global analysis of metabolites in AH has been mainly performed using NMR. Among the analytical platforms used in metabolomics, LC-MS allows for the highest metabolome coverage. The aim of this study was to develop a method for extraction and analysis of AH metabolites by LC-QTOF-MS. Different protocols for AH preparation were tested. The best results were obtained when one volume of AH was mixed with one volume of methanol : ethanol (1 : 1). In the final method, 2 µL of extracted sample was analyzed by LC-QTOF-MS. The method allowed for reproducible measurement of over 1000 metabolic features. Almost 250 metabolites were identified in AH and assigned to 47 metabolic pathways. This method is suitable to study the potential role of amino acids, lipids, oxidative stress, or microbial metabolites in development of ocular diseases.
“…The group of Song et al have studied the metabolic changes in rabbit AH after glucocorticosteroids administration [15] and the hypotensive effects of glycyrrhizin on a rabbit model of ocular hypertension induced by triamcinolone acetonide [16]. 1 H NMR has also been used to study metabolic alterations in AH in the rat glaucoma model induced by intracameral sodium hyaluronate injections [17] and to evaluate the effect of UV-A and UV-B irradiation on the metabolic profile of aqueous humor in rabbits [18]. Currently there is only one metabolomics study on human aqueous humor.…”
Aqueous humor (AH) is a transparent fluid which fills the anterior and posterior chambers of the eye. It supplies nutrients and removes metabolic waste from avascular tissues in the eye. Proper homeostasis of AH is required to maintain adequate intraocular pressure as well as optical and refractive properties of the eye. Application of metabolomics to study human AH may improve knowledge about the molecular mechanisms of eye diseases. Until now, global analysis of metabolites in AH has been mainly performed using NMR. Among the analytical platforms used in metabolomics, LC-MS allows for the highest metabolome coverage. The aim of this study was to develop a method for extraction and analysis of AH metabolites by LC-QTOF-MS. Different protocols for AH preparation were tested. The best results were obtained when one volume of AH was mixed with one volume of methanol : ethanol (1 : 1). In the final method, 2 µL of extracted sample was analyzed by LC-QTOF-MS. The method allowed for reproducible measurement of over 1000 metabolic features. Almost 250 metabolites were identified in AH and assigned to 47 metabolic pathways. This method is suitable to study the potential role of amino acids, lipids, oxidative stress, or microbial metabolites in development of ocular diseases.
“…Aqueous humour from a rat glaucoma model induced by intracameral sodium hyaluronate injection was analysed using NMR in a study by Mayodomo-Febrer et al 58 The study showed a reduced glucose level in the glaucomatous eyes compared with healthy eyes. The authors speculated that increased intraocular pressure could lead to a reduced energy supply to the anterior chamber tissue (reflected by the reduced glucose level), which in turns alters the outflow facility in the trabecular meshwork.…”
Metabolomics is the study of endogenous and exogenous metabolites in biological systems, which aims to provide comparative semi-quantitative information about all metabolites in the system. Metabolomics is an emerging and potentially powerful tool in ophthalmology research. It is therefore important for health professionals and researchers involved in the speciality to understand the basic principles of metabolomics experiments. This article provides an overview of the experimental workflow and examples of its use in ophthalmology research from the study of disease metabolism and pathogenesis to identification of biomarkers.
“…Twenty-one, male, Sprague Dawley rats were included in the post-mortem study (42 eyes in total) and six rats were included in the tissue comparison study; all rats were in the age-range of [7][8][9][10][11][12] weeks. The rats were kept in the same room under standard laboratory conditions and fed with standard chow.…”
Section: Animalsmentioning
confidence: 99%
“…[6][7][8][9][10] It has also been used to study ageing, a variety of ocular pathologies, metabolite transport in the retina, effects of drugs and metabolic changes during corneal organ culture for transplantation. [11][12][13][14][15] Aqueous and vitreous are the most accessible intraocular samples that can be obtained in clinical studies, and these have been used to interrogate the metabolic activities of surrounding ocular structures. 11,15 However, it is unclear whether sampling ocular biofluids provides robust and reproducible information on the metabolic activity of surrounding tissues and ideally, the metabolome of the ocular tissue of interest should be studied directly.…”
Section: Introductionmentioning
confidence: 99%
“…[11][12][13][14][15] Aqueous and vitreous are the most accessible intraocular samples that can be obtained in clinical studies, and these have been used to interrogate the metabolic activities of surrounding ocular structures. 11,15 However, it is unclear whether sampling ocular biofluids provides robust and reproducible information on the metabolic activity of surrounding tissues and ideally, the metabolome of the ocular tissue of interest should be studied directly. One option is to use post-mortem eyes.…”
and 48 hours post-mortem. To study the metabolite composition of rat ocular tissues, eyes were dissected immediately after culling to isolate the cornea, lens, vitreous and retina, prior to storing at -80 o C. Tissue extracts were subjected to Gas Chromatograph Mass Spectrometry (GC-MS) and Ultra High Performance Liquid Chromatography Mass Spectrometry (UHPLC-MS). Generally, the metabolic composition of the retina was stable for 8 hours post-mortem when eyes were stored at 4 o C, but showed increasing changes thereafter. However, some more rapid changes were observed such as increases in TCA cycle metabolites after 2 hours post-mortem, whereas some metabolites such as fatty acids only showed decreases in concentration from 24 hours. A total of 42 metabolites were identified across the ocular tissues by GC-MS (MSI level 1) and 2782 metabolites were annotated by UHPLC-MS (MSI level 2) according to MSI reporting standards. Many of the metabolites detected were common to all of the tissues but some metabolites showed partitioning between different ocular structures with 655, 297, 93 and 13 metabolites being uniquely detected in the retina, lens, cornea and vitreous respectively. Only a small percentage (1.6%) of metabolites found in the vitreous were exclusively found in the retina and not other tissues. In conclusion, mass
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