{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,11]],"date-time":"2025-12-11T20:42:10Z","timestamp":1765485730903,"version":"3.40.4"},"reference-count":43,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2013,11,13]],"date-time":"2013-11-13T00:00:00Z","timestamp":1384300800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"NIH Grants","award":["R21 ES021311","R01 AA018869","P01 AA017103","R01 AA018016","R37 AA010762","P30 AA019360","RC2AA019385","R01 AA015970","R01 DK071765"],"award-info":[{"award-number":["R21 ES021311","R01 AA018869","P01 AA017103","R01 AA018016","R37 AA010762","P30 AA019360","RC2AA019385","R01 AA015970","R01 DK071765"]}]},{"DOI":"10.13039\/100000738","name":"Department of Veterans Affairs","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100000738","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Alcoholism Clin &amp; Exp Res"],"published-print":{"date-parts":[[2014,4]]},"abstract":"Background<\/jats:title>Chronic alcohol exposure results in liver injury that is driven in part by inflammatory cytokines such as tumor necrosis factor\u2010\u03b1 (TNF<\/jats:styled-content>). Hepatocytes are normally resistant to the cytotoxic effects ofTNF<\/jats:styled-content>, but they become sensitized toTNF<\/jats:styled-content>by chronic alcohol exposure. Recently, we reported that the decrease in the ratio ofS<\/jats:styled-content>\u2010adenosylmethionine (SAM<\/jats:styled-content>) toS<\/jats:styled-content>\u2010adenosylhomocysteine (SAH<\/jats:styled-content>) that occurs with alcoholic liver injury renders hepatocytes sensitive toTNF<\/jats:styled-content>cytotoxicity. The purpose of this study was to determine whether inhibition of the transcription factor nuclear factor\u2010kappaB (NF<\/jats:styled-content>\u2010\u03baB<\/jats:styled-content>) contributed toTNF<\/jats:styled-content>\u2010induced cell death in hepatocytes with high levels ofSAH<\/jats:styled-content>.<\/jats:p><\/jats:sec>Methods<\/jats:title>Primary human hepatocytes orH<\/jats:styled-content>epG<\/jats:styled-content>2 cells were pre\u2010incubated with a combination of adenosine plus homocysteine to increaseSAH<\/jats:styled-content>levels. Following exposure toTNF<\/jats:styled-content>, viability was determined by theMTT<\/jats:styled-content>assay, and activation of theNF<\/jats:styled-content>\u2010\u03baB<\/jats:styled-content>pathway was assessed by measuring degradation of cytosolicI<\/jats:styled-content>\u03baB<\/jats:styled-content>\u2010\u03b1, phosphorylation and translocation ofNF<\/jats:styled-content>\u2010\u03baB<\/jats:styled-content>to the nucleus, and expression ofNF<\/jats:styled-content>\u2010\u03baB<\/jats:styled-content>\u2010dependent genes.TNF<\/jats:styled-content>\u2010induced apoptotic signaling pathways were assessed by monitoring levels of the anti\u2010apoptotic protein,A<\/jats:styled-content>20, and cleavage products of the caspase\u20108 substrate,RIP<\/jats:styled-content>1.<\/jats:p><\/jats:sec>Results<\/jats:title>NF<\/jats:styled-content>\u2010\u03baB<\/jats:styled-content>\u2010mediated gene expression was inhibited in cells with highSAH<\/jats:styled-content>, despite the fact thatTNF<\/jats:styled-content>\u2010induced degradation of the cytoplasmic inhibitorI<\/jats:styled-content>\u03baB<\/jats:styled-content>\u2010\u03b1 and accumulation ofNF<\/jats:styled-content>\u2010\u03baB<\/jats:styled-content>in the nucleus persisted for much longer. In contrast to control cells, theNF<\/jats:styled-content>\u2010\u03baB<\/jats:styled-content>that accumulated in the nucleus of cells with highSAH<\/jats:styled-content>levels was not phosphorylated at serine 536, a modification associated with activation of the transactivation potential of this transcription factor. The inhibition of transactivation byNF<\/jats:styled-content>\u2010\u03baB<\/jats:styled-content>resulted in lowermRNA<\/jats:styled-content>and protein levels of the anti\u2010apoptotic proteinA<\/jats:styled-content>20 and increased cleavage ofRIP<\/jats:styled-content>1.<\/jats:p><\/jats:sec>Conclusions<\/jats:title>HighSAH<\/jats:styled-content>levels inhibitedNF<\/jats:styled-content>\u2010\u03baB<\/jats:styled-content>\u2010mediated gene expression and sensitized primary hepatocytes andH<\/jats:styled-content>epG<\/jats:styled-content>2 cells to the cytotoxic effects ofTNF<\/jats:styled-content>. It is likely that crosstalk with other transcription factors is perturbed under these conditions, resulting in still other changes in gene expression.<\/jats:p><\/jats:sec>","DOI":"10.1111\/acer.12315","type":"journal-article","created":{"date-parts":[[2013,11,13]],"date-time":"2013-11-13T17:39:55Z","timestamp":1384364395000},"page":"889-896","source":"Crossref","is-referenced-by-count":8,"title":["S\u2010Adenosylhomocysteine InhibitsNF<\/scp>\u2010\u03baB<\/scp>\u2010Mediated Gene Expression in Hepatocytes and Confers Sensitivity toTNF<\/scp>Cytotoxicity"],"prefix":"10.1111","volume":"38","author":[{"given":"Walter H.","family":"Watson","sequence":"first","affiliation":[{"name":"Division of Gastroenterology, Hepatology and Nutrition Department of Medicine University of Louisville School of Medicine Louisville Kentucky"},{"name":"University of Louisville Alcohol Research Center Louisville Kentucky"},{"name":"Department of Pharmacology and Toxicology University of Louisville School of Medicine Louisville Kentucky"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tom J.","family":"Burke","sequence":"additional","affiliation":[{"name":"Division of Gastroenterology, Hepatology and Nutrition Department of Medicine University of Louisville School of Medicine Louisville Kentucky"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mark A.","family":"Doll","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Toxicology University of Louisville School of Medicine Louisville Kentucky"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Craig J.","family":"McClain","sequence":"additional","affiliation":[{"name":"Division of Gastroenterology, Hepatology and Nutrition Department of Medicine University of Louisville School of Medicine Louisville Kentucky"},{"name":"University of Louisville Alcohol Research Center Louisville Kentucky"},{"name":"Department of Pharmacology and Toxicology University of Louisville School of Medicine Louisville Kentucky"},{"name":"Robley Rex Veterans Affairs Medical Center Louisville Kentucky"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2013,11,13]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1053\/jhep.2002.31309"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.274.5288.782"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1158\/2159-8290.CD-11-0172"},{"key":"e_1_2_6_5_1","doi-asserted-by":"crossref","first-page":"1736","DOI":"10.4049\/jimmunol.153.4.1736","article-title":"Adenosine and homocysteine together enhance TNF\u2010mediated cytotoxicity but do not alter activation of nuclear factor\u2010kappa B in L929 cells","volume":"153","author":"Bergmann S","year":"1994","journal-title":"J 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