Cystathionine β Synthase Deficiency Promotes Oxidative Stress, Fibrosis, and Steatosis in Mice Liver

doi:10.1053/j.gastro.2005.02.034

Gastroenterology

Volume 128, Issue 5, May 2005, Pages 1405-1415

https://doi.org/10.1053/j.gastro.2005.02.034 Get rights and content

Background & Aims: Cystathionine β-synthase (CBS) deficiency causes severe hyperhomocysteinemia, which confers diverse clinical manifestations, notably liver disease. To investigate this aspect of hyperhomocysteinemia, we performed a thorough investigation of liver pathology in CBS-deficient mice, a murine model of severe hyperhomocysteinemia. Methods: The degree of liver injury and inflammation was assessed by histologic examination, by measurements of products of lipid peroxidation, and by formation of carbonyl groups on protein as a measure for the occurrence of protein oxidation. Analysis of profibrogenic, proinflammatory factors and cell apoptosis was performed by Western blots, real-time quantitative reverse-transcription polymerase chain reaction, caspase-3 activity, DNA laddering, and TUNEL assay. Results: Histologic evaluation of liver specimens of 8- to 32-week-old CBS-deficient mice showed that CBS-deficient mice develop inflammation, fibrosis, and hepatic steatosis, concomitant with an enhanced expression of tissue inhibitor of metalloproteinase-1, α-smooth muscle actin, pro(α)1 collagen type I, transforming growth factor-β1, and proinflammatory cytokines. Moreover, even if the proapoptotic protein Bax was dominantly expressed and Bcl-2 was down-regulated, caspase-3 was not activated, DNA laddering was not detected, and number of positive TUNEL cells was not increased in liver of CBS-deficient mice compared with wild-type mice. Conclusions: The results show that hyperhomocysteinemia in liver of CBS-deficient mice promotes oxidative stress, which may cause mitochondrial damage in association with activation of hepatic stellate cells, leading to liver injury. The absence of caspase-3 activation, DNA fragmentation, and TUNEL-positive cells shows that protective signals may counteract apoptotic signals in liver of CBS-deficient mice.

Section snippets

Animals and Experimental Protocols

Heterozygous CBS-deficient (CBS +/−) mice were generously donated by Dr. N. Maeda (Department of Pathology, University of North Carolina, Chapel Hill, NC) and maintained in a controlled environment with access to unlimited food and water. All animals received human care according to the criteria outlined in the guide for the Care and Use of Laboratory Animals. CBS +/− mice were bred to obtain homozygous CBS-deficient (CBS −/−) mice. This produced CBS −/− and wild-type (CBS +/+) mice from the

Total Plasma and Hepatic Homocysteine Levels in CBS-Deficient Mice

Plasma tHcy concentration was 50-fold higher in CBS −/− mice, which is considered as a murine model of severe hyperhomocysteinemia,⁶ than that in CBS +/+ mice (205 ± 86 vs. 3.9 ± 0.9 μmol/L, respectively; P < .0001 by Student t test [n = 4 for each]). The mean hepatic concentration of tHcy in CBS −/− mice was approximately 20-fold higher than that in CBS +/+ mice (tHcy concentrations were 6.44 ± 3.86 vs. 0.34 ± 0.14 nmol/mg of cellular protein, respectively; P < .001 by Student t test [n = 5

Discussion

Despite clinical evidence for the toxicity of excess Hcy, the specific cytopathologic mechanisms of hepatocellular injury because of hyperhomocysteinemia have not been fully explored. The present study provides evidence that hyperhomocysteinemia because of CBS deficiency in mice was associated with increased hepatic oxidative stress. Enhanced lipid peroxidation seems to be a key feature in the liver injury observed in CBS-deficient mice. The process of lipid peroxidation generates numerous

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: Supported by the European Union Grant (QLRT-2001-00816), a grant “vin et santé” and the Fondation de France, a fellowship from the Fondation pour la Recherche Médicale (FRM) and the GEHT-ISTH 2001 (to K.R.), and a fellowship from the Société Française d’Hypertension Artérielle (SFHTA; to J.N.).

¹: K.R. and J.N. contributed equally to this paper.

²: E.B.’s current address is EA 2526, Laboratoire de Biochimie et Génétique Moléculaire; Faculté des Sciences et Technologies, Université de la Réunion, 15, Avenue René Cassin, BP 7151; 97715 Saint Denis Messag Cedex 09, La réunion, France.

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Basic-liver, pancreas, and biliary tractCystathionine β Synthase Deficiency Promotes Oxidative Stress, Fibrosis, and Steatosis in Mice Liver

Section snippets

Animals and Experimental Protocols

Total Plasma and Hepatic Homocysteine Levels in CBS-Deficient Mice

Discussion

J Biol Chem

Biochem Biophys Res Commun

Am J Gastroenterol

J Biol Chem

J Lab Clin Med

J Biol Chem

Thromb Res

J Nutr

Free Radic Biol Med

J Biol Chem

J Nutr

J Biol Chem

J Lipid Res

Gastroenterology

Biochim Biophys Acta

Gastroenterology

Hepatology

Atherosclerosis

J Biol Chem

Hepatology

J Biol Chem

J Hepatol

Hepatol Res

J Hepatol

Hepatology

Disorders of transsulfuration

Homocysteine metabolism

Annu Rev Nutr

Homocysteine and atherothrombosis

N Engl J Med

Basic-liver, pancreas, and biliary tract
Cystathionine β Synthase Deficiency Promotes Oxidative Stress, Fibrosis, and Steatosis in Mice Liver