High-Resolution Magic Angle Spinning 1H NMR Spectroscopy of Intact Liver and Kidney: Optimization of Sample Preparation Procedures and Biochemical Stability of Tissue during Spectral Acquisition

Waters, Nigel J.; Garrod, S.; Farrant, R. D.; Haselden, John N.; Connor, Susan C.; Connelly, John C.; Lindon, John C.; Holmes, Elaine; Nicholson, Jeremy K.

doi:10.1006/abio.2000.4574

Cited by 121 publications

(110 citation statements)

References 19 publications

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“…Spherical inserts were used in all the cases, limiting the rotor inner volume to 12 μl. HR-MAS using spherical small volume inserts provides improved resolution and line shapes compared to cylindrical inserts (40). D 2 O exceeding the 12 μl volume was removed before rotor sealing.…”

Section: Methodsmentioning

confidence: 99%

Metabolic profile of chronic liver disease by NMR spectroscopy of human biopsies

Monleνn

2010

Int J Mol Med

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Abstract. Among the different processes occurring during the evolution of liver disease, fibrosis has a predominant role. Liver fibrosis mechanisms are fairly constant irrespective of the underlying etiology. Cirrhosis is the end-stage of this reaction. Metabolic profiles, which are affected by many physiological and pathological processes, may provide further insight into the metabolic consequences of this severe liver disease. The aim of this study was to demonstrate the applicability of 1 H high resolution magic angle spinning (HR-MAS) NMR spectroscopy in the biochemical profile determination of human liver needle biopsy samples for the characterization of metabolic alterations related to the severity of liver disease. We recorded and analyzed HR-MAS spectra of 68 liver tissue samples obtained by needle biopsy from patients with chronic liver disease. Multivariate analysis was applied to these data to obtain discrimination patterns and to reveal relevant metabolites. The metabolic characterization of liver tissue from needle biopsies by HR-MAS NMR spectroscopy provided differential patterns for cirrhotic and non-cirrhotic chronic liver disease tissue. Metabolites closely related to the liver metabolism such as some fatty acids, glucose and amino acids show differences between the two groups. Phospholipid precursors, which have been previously correlated with hepatic lesions also show differences. Furthermore, the correlation between histologically assessed liver disease stages and the levels of the most discriminative metabolites show that liver dysfunction is present at the initial stages of chronic hepatic lesions. Overall, this work suggests that the additional information obtained by NMR metabolomics applied to needle biopsies of human liver may be useful for assessing metabolic alterations and liver dysfunction in chronic liver disease. IntroductionThe liver is among the most metabolically diverse organs of the body and is involved in many critical metabolic processes (1). One of its major roles is storing endogenous fuels of the body. In addition, the liver acts as a major distribution center for exogenous energy supplies. The liver may be affected by many pathological aggressions. Liver dysfunction results in a variety of clinical signs and symptoms. However, some of these pathologies pose a diagnostic challenge as many different diseases show similar clinical signs (2-4). Moreover, liver biopsy assessment involves some degree of complication because of the proliferation of different grading and staging schemes (5-7). Metabolic alterations produced by liver disease may help clinicians better characterize the disease. Among the different processes occurring during the evolution of liver disease, fibrosis takes a predominant role. Liver fibrosis is characterized by the replacement of liver tissue by fibrous scar tissue as well as regenerative nodules, leading to progressive loss of liver function and to altered liver metabolism (8-11). Liver fibrosis induction mechanisms are fairly constant irrespectivel...

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Section: Methodsmentioning

confidence: 99%

Metabolic profile of chronic liver disease by NMR spectroscopy of human biopsies

Monleνn

2010

Int J Mol Med

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“…Due to the limited size of the adrenal glands, tissue extracts were prepared from both glands following the methods previously described. 22 Tissue samples were ground in 1 mL of acetonitrile/water (1:1). The supernatant containing the aqueous phase was collected, freeze-dried and redissolved in 60 µL of D 2 O.…”

Section: Methodsmentioning

confidence: 99%

“…22 Tissue samples were regulated at 283 K using cold N 2 gas to minimize any time-dependent biochemical degradation.…”

Section: Methodsmentioning

confidence: 99%

Panorganismal Gut Microbiome−Host Metabolic Crosstalk

et al. 2009

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Coevolution shapes interorganismal crosstalk leading to profound and diverse cellular and metabolic changes as observed in gut dysbiosis in human diseases. Here, we modulated a simplified gut microbiota using pro-, pre-, and synbiotics to assess the depth of systemic metabolic exchanges in mice, using a multicompartmental modeling approach with metabolic signatures from 10 tissue/fluid compartments. The nutritionally induced microbial changes modulated host lipid, carbohydrate, and amino acid metabolism at a panorganismal scale. Galactosyl-oligosaccharides reduced lipogenesis, triacylglycerol incorporation into lipoproteins and triglyceride concentration in the liver and the kidney. Those changes were not correlated with decreased plasma lipoproteins that were specifically induced by L. rhamnosus supplementation. Additional alteration of transmethylation metabolic pathways (homocysteine-betaine) was observed in the liver and the pancreas following pre-and synbiotic microbial modulation, which may be of interest for control of glucose metabolism and insulin sensitivity. Probiotics also reduced hepatic glycogen and glutamine and adrenal ascorbate with inferred effects on energy homeostasis, antioxidation, and steroidogenesis. These studies show the breadth and the depth of gut microbiome modulations of host biochemistry and reveal that major mammalian metabolic processes are under symbiotic homeostatic control.

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“…As a consequence, MRS is well suited for in vivo, longitudinal studies of liver lipids in rodents. Proton MRS methods have been used widely to characterize triacylglycerols and other fatty acids in tissue and tissue extracts (27)(28)(29)(30), and numerous MRS studies of liver lipids in humans have been described (18,(31)(32)(33)(34). MRS measurements of liver fat levels in rats have been reported previously (35,36).…”

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confidence: 99%

In vivo MRS measurement of liver lipid levels in mice

Garbow

Lin

Sakata

et al. 2004

Journal of Lipid Research

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A magnetic resonance spectroscopy (MRS) procedure for in vivo measurement of lipid levels in mouse liver is described and validated. The method uses respiratory-gated, localized spectroscopy to collect proton spectra from voxels within the mouse liver. Bayesian probability theory analysis of these spectra allows the relative intensities of the lipid and water resonances within the liver to be accurately measured. All spectral data were corrected for measured spin-spin relaxation. A total of 48 mice were used in this study, including wild-type mice and two different transgenic mouse strains. Different groups of these mice were fed high-fat or low-fat diets or liquid diets with and without the addition of alcohol. Proton spectra were collected at baseline and, subsequently, Alcoholic (1, 2) and nonalcoholic fatty liver (NAFL) (3-7) are highly prevalent in human populations and may develop into steatohepatitis and in some cases into cirrhosis requiring liver transplantation. Animal models of both conditions have been developed. Mouse models are particularly useful because genetic manipulations are highly developed in inbred mice. However, longitudinal studies require the killing of animals because no noninvasive method for quantifying liver fat is available. Here, we describe a noninvasive, nondestructive method for quantifying the liver fat contents of the mouse using a NAFL mouse model. The overwhelming majority of NAFL cases are associated with obesity, dyslipidemia, hypertension, insulin-resistant type 2 diabetes mellitus, and atherosclerotic cardiovascular disease (8)(9)(10)(11)(12). This constellation defines the metabolic syndrome (13,14). One naturally occurring cause of fatty liver is familial hypobetalipoproteinemia (FHBL). FHBL is defined by less than fifth percentile plasma levels of LDL-cholesterol and/or total apolipoprotein B (apoB), segregating in families as an autosomal dominant trait (15-17). The mean liver triglyceride content in apoBimpaired FHBL subjects is 3-to 5-fold greater than that of controls (18,19).In an attempt to understand the cellular/molecular bases of hypobetalipoproteinemia, several recombinant mice mimicking human FHBL have been produced (19)(20)(21)(22)(23)(24)(25). The resulting mice closely resemble their human counterparts with respect to the fatty liver phenotype. Thus, these mice could serve as good models of one genetic form of fatty liver for studies of the progression of fatty liver and on the effects of metabolic, hormonal, and therapeutic perturbations over time.One current limitation of such studies is the absence of an accurate, quantifiable, noninvasive method for repeat assessments of liver fat in animals. In humans, the gold standard for quantifying liver fat is the liver biopsy, which is invasive and, at best, semiquantitative. The analogous procedure in animals is killing of the animal and direct analysis of liver fat by standard chemical methods after extraction. Obviously, direct sampling of livers for fat analysis is not ideal for longitudinal, fol...

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High-Resolution Magic Angle Spinning 1H NMR Spectroscopy of Intact Liver and Kidney: Optimization of Sample Preparation Procedures and Biochemical Stability of Tissue during Spectral Acquisition

Cited by 121 publications

References 19 publications

Metabolic profile of chronic liver disease by NMR spectroscopy of human biopsies

Metabolic profile of chronic liver disease by NMR spectroscopy of human biopsies

Panorganismal Gut Microbiome−Host Metabolic Crosstalk

In vivo MRS measurement of liver lipid levels in mice

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