Mitochondria are the major targets in albumin-induced apoptosis in proximal tubule cells.
Nephrotic-range proteinuria is considered a poor prognostic factor. A correlation between tubulointerstitial injury and the degree of proteinuria is well established. In an attempt to explain the tubular atrophy that is observed in advanced glomerulonephritides, this study investigated apoptotic mechanisms in cultured human proximal tubule cells (HKC-8) that were exposed to endotoxin-free albumin (5, 10, and 20 mg/ml). Apoptosis was detected by Hoechst 33342; annexin staining; and assays for caspases 3, 8, and 9. The apoptotic effect of albumin was maximal at 10 mg/ml albumin, and necrosis prevailed in cells that were incubated with 20 mg/ml. Increase in caspase-9 and -3 activity was observed starting at 6 and maximally at 16 to 24 h. The proapoptotic Bcl-2 protein Bax was upregulated at 6 h, associated with translocation of cytochrome-c from mitochondria to cytosol and alteration in the mitochondrial membrane potential. Production of reactive oxygen species (ROS) was significant at 6 h but declined at 16 and 24 h. Treatment with ROS scavenger dimethylthiourea or antioxidant N-acetylcysteine did not alleviate caspase-3 production. Pan protein kinase C inhibitor bisindolylmaleimide-1 protected the cells from apoptosis. It is concluded that albumin induces apoptosis in human proximal tubule cells by stimulating mitochondrial apoptotic pathway independent of ROS production.
Erkan
Elif
E
Department of Pediatrics, University of Rochester, School of Medicine, Rochester, New York, USA. elif.erkan@chp.edu
Devarajan
Prasad
P
Schwartz
George J
GJ
D000111:Acetylcysteine / Q000494:pharmacology; D000418:Albumins / Q000633:toxicity*; D017209:Apoptosis / Q000187:drug effects*; D053453:Caspase 9 / Q000502:physiology; D002460:Cell Line; D045304:Cytochromes c / Q000378:metabolism; D006801:Humans; D007687:Kidney Tubules, Proximal / Q000187:drug effects* / Q000473:pathology; D026561:Low Density Lipoprotein Receptor-Related Protein-2 / Q000502:physiology; D008564:Membrane Potentials / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology; D021381:Protein Transport; D017382:Reactive Oxygen Species / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000378:metabolism
D000418:Albumins; D026561:Low Density Lipoprotein Receptor-Related Protein-2; D017382:Reactive Oxygen Species; D051028:bcl-2-Associated X Protein; D045304:Cytochromes c; D011493:Protein Kinase C; D053453:Caspase 9; D000111:Acetylcysteine