Prevention of cold ischemia/rewarming-induced ERK 1/2, p38 kinase and HO-1 activation by trophic factor supplementation of UW solution.
We have previously shown that trophic factor supplementation (TFS) of University of Wisconsin (UW) solution reduced early apoptotic changes in vascular endothelial cells. Here, we examine the effect of TFS on cell signaling pathways related to cell growth, differentiation, and apoptosis after cold ischemic storage. In this study, the effect of TFS on the phosphorylation of signaling molecules ERK (extracellular regulated-signaling kinase) 1/2 and p38 MAPK (mitogen activated protein kinases) and of HO-1 (hemeoxygenase-1), relative to changes seen in unmodified UW solution, were determined by Western blot in cells stored under cold ischemic conditions. Primary cultures of canine kidney proximal tubule cells (CKPTC) and human umbilical vein endothelial cells (HUVEC) were used in this study. There was a significant decrease, relative to UW solution, after 1 min rewarming in ERK 1 and 2 activity in CKPTCs. For p38 MAPK, a significant decrease after 5 min rewarming was seen in CKPTC (p<0.05) while significant reductions relative to UW solution were seen in HUVECs after both 1 and 5 min rewarming (p<0.05). Phosphorylated HO-1 was also decreased by 43% and 50% in HUVECs, relative to UW solution, after 1 and 5 min rewarming (p<0.05 at each time point). Collectively, TFS not only limits ERK 1/2 and p38 MAPK activity induced by cold ischemic injury and subsequent rewarming, but also substantially restricted increases in HO-1 phosphorylation.
Kwon
Young Sam
YS
Department of Surgical Sciences, School of Veterinary Medicine, University of Wisconsin, 2015 Linden Drive W, Madison, WI 53706-1102, USA.
Foley
John D
JD
Russell
Paul
P
McAnulty
Jonathan F
JF
Murphy
Christopher J
CJ
D000241:Adenosine / Q000378:metabolism / Q000494:pharmacology; D000493:Allopurinol / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis; D002478:Cells, Cultured; D050377:Cold Ischemia; D004285:Dogs; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism / Q000494:pharmacology; D051547:Heme Oxygenase-1 / Q000378:metabolism*; D006801:Humans; D007328:Insulin / Q000378:metabolism / Q000494:pharmacology; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D007684:Kidney Tubules / Q000166:cytology / Q000187:drug effects; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D019188:Organ Preservation Solutions / Q000378:metabolism / Q000494:pharmacology*; D010766:Phosphorylation; D011887:Raffinose / Q000378:metabolism / Q000494:pharmacology; D015398:Signal Transduction; D014922:Wisconsin; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism*
D007328:Insulin; D036341:Intercellular Signaling Peptides and Proteins; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; D000493:Allopurinol; D051547:Heme Oxygenase-1; D048049:Extracellular Signal-Regulated MAP Kinases; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D048051:p38 Mitogen-Activated Protein Kinases; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose