Expression of intestinal brush-border membrane hydrolases and ferritin after segmental ischemia-reperfusion in rats.
Jejunal expression of three brush-border membrane (BBM) enzymes, intestinal alkaline phosphatase (IAP), lactose-phlorizin hydrolase (LPH), and sucrase-isomaltase (SI), and a cytosolic protein, ferritin (Ft), was investigated after transient segmental ischemia-reperfusion (I/R). I/R reduced mucosal IAP, LPH, and SI mRNAs to 36%, 11%, and 38% of normal jejunal levels after 3 h of reperfusion and to 22%, 8%, and 51% of normal jejunal levels after 6 h of reperfusion, respectively. Intriguingly, in the internal control jejunum IAP and LPH mRNAs also decreased significantly. LPH and SI mRNA rapidly recovered to levels significantly higher than those of normal jejunum at 12 h, whereas IAP mRNA levels did not recover until 48 h. Enzyme activity paralleled changes in mRNA levels in the ischemic reperfused jejunum. Electrophoretic mobility shift assays showed that I/R significantly increased SI footprinting 1 (SIF1) binding activity. The mobility of one of the DNA-protein complexes was further retarded in the presence of anti-Cdx-2 antibody, suggesting that either Cdx-2 or a related protein was interacting with the SIF1 sequences. Similar to BBM enzymes, cytosolic Ft mRNA and protein were significantly decreased at 3 and 6 h after I/R. By 12 h, Ft mRNA, but not Ft protein, had increased to higher than normal levels. We conclude that a rapid recovery of BBM mRNAs and enzymes occurs in regenerating mucosa after upper villus damage. The increase of SIF1 binding protein activity after I/R may enhance SI, and perhaps LPH, gene transcription. The expression of Ft is regulated at both pretranslational and translational levels.
Department of Medicine, Feist-Weiller Cancer Center, Louisiana State University Medical Center, Shreveport, Louisiana 71130, USA.
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D000469:Alkaline Phosphatase / Q000235:genetics; D000818:Animals; D003600:Cytosol / Q000378:metabolism; D005293:Ferritins / Q000096:biosynthesis / Q000235:genetics*; D019084:Fluorescent Antibody Technique, Indirect; D005786:Gene Expression Regulation; D006867:Hydrolases / Q000096:biosynthesis / Q000235:genetics*; D007413:Intestinal Mucosa / Q000098:blood supply* / Q000378:metabolism* / Q000473:pathology; D007511:Ischemia / Q000378:metabolism* / Q000473:pathology; D007583:Jejunum / Q000098:blood supply*; D007700:Kinetics; D010694:Lactase-Phlorizin Hydrolase / Q000235:genetics; D008871:Microvilli / Q000378:metabolism* / Q000473:pathology / Q000648:ultrastructure; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D015424:Reperfusion; D013394:Sucrase-Isomaltase Complex / Q000235:genetics; D013997:Time Factors; D014158:Transcription, Genetic
D012333:RNA, Messenger; D005293:Ferritins; D006867:Hydrolases; D000469:Alkaline Phosphatase; D013394:Sucrase-Isomaltase Complex; D010694:Lactase-Phlorizin Hydrolase