title: Studies on hepatic injury and antioxidant enzyme activities in rat subcellular organelles following in vivo ischemia and reperfusion.
abstract: The activities of rat hepatic subcellular antioxidant enzymes were studied during hepatic ischemia/reperfusion. Ischemia was induced for 30 min (reversible ischemia) or 60 min (irreversible ischemia). Ischemia was followed by 2 or 24 h of reperfusion. Hepatocyte peroxisomal catalase enzyme activity decreased during 60 min of ischemia and declined further during reperfusion. Peroxisomes of normal density (d = 1.225 gram/ml) were observed in control tissues. However, 60 min of ischemia also produced a second peak of catalase specific activity in subcellular fractions corresponding to newly formed low density immature peroxisomes (d = 1.12 gram/ml). The second peak was also detectable after 30 min of ischemia followed by reperfusion for 2 or 24 h. Mitochondrial and microsomal fractions responded differently. MnSOD activity in mitochondria and microsomal fractions increased significantly (p < 0.05) after 30 min of ischemia, but decreased below control values following 60 min of ischemia and remained lower during reperfusion at 2 and 24 h in both organelle fractions. Conversely, mitochondrial and microsomal glutathione peroxidase (GPx) activity increased significantly (p < 0.001) after 60 min of ischemia and was sustained during 24 h of reperfusion. In the cytosolic fraction, a significant increase in CuZnSOD activity was noted following reperfusion in animals subjected to 30 min of ischemia, but 60 min of ischemia and 24 h of reperfusion resulted in decreased CuZnSOD activity. These studies suggest that the antioxidant enzymes of various subcellular compartments respond to ischemia/reperfusion in an organelle or compartment specific manner and that the regulation of antioxidant enzyme activity in peroxisomes may differ from that in mitochondria and microsomes. The compartmentalized changes in hepatic antioxidant enzyme activity may be crucial determinant of cell survival and function during ischemia/reperfusion. Finally, a progressive decline in the level of hepatic reduced glutathione (GSH) and concomitant increase in serum glutamate pyruvate transaminase (SGPT) activity also suggest that greater tissue damage and impairment of intracellular antioxidant activity occur with longer ischemia periods, and during reperfusion.
authors.affiliation: Department of Pediatrics, Medical University of South Carolina, Charleston 29425, USA.
authors.forename: M
authors.initials: M
authors.lastname: Gupta
authors.forename: K
authors.initials: K
authors.lastname: Dobashi
authors.forename: E L
authors.initials: EL
authors.lastname: Greene
authors.forename: J K
authors.initials: JK
authors.lastname: Orak
authors.forename: I
authors.initials: I
authors.lastname: Singh
mesh_terms: D000410:Alanine Transaminase / Q000378:metabolism; D000818:Animals; D002374:Catalase / Q000378:metabolism; D003576:Electron Transport Complex IV / Q000378:metabolism; D005979:Glutathione Peroxidase / Q000378:metabolism; D005980:Glutathione Reductase / Q000378:metabolism; D007511:Ischemia / Q000201:enzymology*; D008099:Liver / Q000098:blood supply* / Q000201:enzymology; D008297:Male; D009245:NADH Dehydrogenase / Q000378:metabolism; D010088:Oxidoreductases / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000201:enzymology*; D013347:Subcellular Fractions / Q000201:enzymology*; D013482:Superoxide Dismutase / Q000378:metabolism
chemical_list: D010088:Oxidoreductases; D002374:Catalase; D005979:Glutathione Peroxidase; D013482:Superoxide Dismutase; D009245:NADH Dehydrogenase; D005980:Glutathione Reductase; D003576:Electron Transport Complex IV; D000410:Alanine Transaminase