Ammonia-induced heme oxygenase-1 expression in cultured rat astrocytes and rat brain in vivo.
Ammonia is a key factor in the pathogenesis of hepatic encephalopathy (HE), which is a major complication in acute and chronic liver failure and other hyperammonemic states. The molecular mechanisms underlying ammonia neurotoxicity and the functional consequences of ammonia on gene expression in astrocytes are incompletely understood. Using cDNA array hybridization technique we identified ammonia as a trigger of heme oxygenase-1 (HO-1) mRNA levels in cultured rat astrocytes. As shown by Northern and Western blot analysis, HO-1 mRNA levels were upregulated by ammonia (0.1-5 mmol/L) after 24 h and protein expression after 72 h in astrocytes. These ammonia effects on HO-1 are probably triggered to a minor extent by ammonia-induced glutamine synthesis or by astrocyte swelling, because HO-1 expression was not inhibited by the glutamine synthetase inhibitor methionine sulfoximine (which abrogated ammonia-induced cell swelling in cultured astrocytes), and ammonia-induced HO-1 expression could only partly be mimicked by hypoosmotic astrocyte swelling. Hypoosmotic (205 mOsm/L) exposure of astrocytes led even to a decrease in HO-1 mRNA levels within 4 h, whereas hyperosmotic (405 mOsm/L) exposure increased HO-1 mRNA expression. After 24 h, hypoosmolarity slightly raised HO-1 mRNA expression. Taurine and melatonin diminished ammonia-induced HO-1 mRNA or protein expression, whereas other antioxidants (dimethylthiourea, butylated hydroxytoluene, N-acetylcysteine, and reduced glutathione) increased HO-1 mRNA levels under ammonia-free conditions. An in vivo relevance is suggested by the finding that increased HO-1 expression occurs in the brain cortex from acutely ammonia-intoxicated rats. It is concluded that ammonia-induced HO-1 expression may contribute to cerebral hyperemia in hyperammonic states.
Warskulat
Ulrich
U
Clinic for Gastroenterology, Hepatology and Infectiology, Heinrich Heine University, Düsseldorf, Germany. warskulat@med.uni-duesseldorf.de
Görg
Boris
B
Bidmon
Hans-J
HJ
Müller
Hans Werner
HW
Schliess
Freimut
F
Häussinger
Dieter
D
D000641:Ammonia / Q000378:metabolism / Q000633:toxicity*; D000818:Animals; D000831:Animals, Newborn; D000975:Antioxidants / Q000494:pharmacology; D001253:Astrocytes / Q000187:drug effects* / Q000201:enzymology*; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000187:drug effects* / Q000201:enzymology* / Q000503:physiopathology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000235:genetics; D005974:Glutamate-Ammonia Ligase / Q000037:antagonists & inhibitors / Q000378:metabolism; D005973:Glutamine / Q000096:biosynthesis; D006419:Heme Oxygenase (Decyclizing) / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D051547:Heme Oxygenase-1; D006501:Hepatic Encephalopathy / Q000201:enzymology* / Q000235:genetics / Q000503:physiopathology; D006940:Hyperemia / Q000201:enzymology / Q000503:physiopathology; D006982:Hypertonic Solutions / Q000494:pharmacology; D007038:Hypotonic Solutions / Q000494:pharmacology; D009997:Osmotic Pressure / Q000187:drug effects; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics
D000975:Antioxidants; D006982:Hypertonic Solutions; D007038:Hypotonic Solutions; D012333:RNA, Messenger; D005973:Glutamine; D000641:Ammonia; D006419:Heme Oxygenase (Decyclizing); D051547:Heme Oxygenase-1; D005974:Glutamate-Ammonia Ligase