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M. Nessim, R.R. Sivaraj, S.P. Young, F. Fortune, M.R. Stanford, P.I. Murray, G.R. Wallace; Metabonomic Analysis of Serum From Patients With Behcet’s Disease: A New Research Tool? . Invest. Ophthalmol. Vis. Sci. 2005;46(13):5098.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Global profiling of the network of proteins or metabolites found in cells, tissues or fluids can increase our understanding of the multiple interacting processes involved in complex systems. One such approach is metabonomics, which we have applied to analyse metabolite profiles in serum of patients with Behçet’s disease (BD), using high–resolution 1H–nuclear magnetic resonance spectroscopy in conjunction with principal component analysis (PCA). Methods: Serum samples from 30 patients with BD were diluted 1:1 with D2O and 1D 1H spectra acquired using a standard pulse sequence with water suppression on a Bruker DRX 500MHz NMR spectrometer equipped with a cryoprobe. Spectra were normalised to a TMSP standard and segmented into 0.005–ppm (2.5 Hz) chemical shift ‘bins’ between 0.2 and 10.0 ppm, and the spectral area within each bin was integrated. Bins between 4.8 and 4.9 ppm containing residual water were removed. PCA of the pre–processed data was conducted using PLS_Toolbox (Eigenvector Research) within MATLAB. Disease activity and drug treatment were recorded and age and sex matched healthy controls were used for comparison. Results: PCA analysis based primarily on lipid (around 1 to 2 ppm) and more hydrophobic (2 to 4ppm) metabolites gave a clear stratification between patients and healthy controls. A distinct separation could also be shown between BD patients and disease controls. There was a distribution associated with patients on and off treatment, and with disease activity. Conclusions: Metabonomic analysis of serum samples shows promise to discriminate between disease activity and response to drug treatment, and may be a valuable research tool in global metabolite profiling in biological samples from patients with BD.
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