A previous study on the natural course of WD in 24-week-old LEC rats showed reduced SAHH protein levels only in advanced stages of liver disease,27 in contrast to findings in the present study of the same age tx-j mice. SAH is a potent inhibitor of DNMTs activities GSK126 and reduced levels of Dnmts transcripts were described in response to elevated SAH levels in a recent clinical study.28 Notably, we observed that Dnmt1, which encodes the principal DNMT in mammalian cells and is mainly implicated in the maintenance of methylation status, was up-regulated, whereas Dnmt3b, encoding a de novo methyltransferase, was down-regulated in untreated tx-j mice, and both

Dnmt3a and Dnmt3b transcripts were down-regulated by PCA. Transcript levels of Dnmt genes show compensatory increased levels in response to reduced DNA methylation in liver or brain.29 Others described up-regulation of Dnmt1 expression without changes in Dnmt3a in livers of choline-deficient rat embryos with global DNA hypomethylation,30

whereas Dnmts levels are increased in chronic hepatitis, cirrhosis, and human hepatocellular carcinoma.31 The finding that Dnmt3b was down-regulated in untreated tx-j mice with increased SAH levels and correlated with global DNA methylation and expressions of each of the studied genes suggests that this enzyme plays a major role in controlling global DNA methylation in WD. There were some discrepancies in the Dnmts expression in response to betaine find more treatment. We did not observe any change in Dnmts levels in control mice nor in Dnmt1 and Dnmt3a in tx-j mice, whereas

Dnmt3b was markedly up-regulated in tx-j mice in response to betaine. Tx-j mice demonstrated reduced levels of global DNA methylation, which was restored either by PCA-induced Cu chelation or by provision check details of the methyl donor betaine. The positive effect of PCA on increasing global DNA methylation may be due to relatively reduced inflammation and demand for methyl groups.17 At the same time, methyl group supplementation by betaine could increase global DNA methylation in both control and tx-j mice, regardless of inflammation. We propose that global DNA hypomethylation in WD was influenced by inflammation which was resolved by reducing Cu hepatic levels by PCA treatment, or by the increasing availability of methyl groups by betaine. The major difference between PCA and betaine treatment in tx-j mice was that PCA was exclusively associated with a significant improvement of inflammation, whereas only betaine induced significant increased SAM and Dnmt3b levels. These observations suggest a potential positive additive effect of including betaine in the PCA treatment of patients with WD.