Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 4 de 4
Filter
Add more filters










Database
Language
Publication year range
1.
Am J Nephrol ; 28(1): 14-24, 2008.
Article in English | MEDLINE | ID: mdl-17890854

ABSTRACT

AIM: The goal of this work is to understand the cellular effects of advanced glycation end product (AGE)-modified protein on renal proximal tubule cells. BACKGROUND: A major function of the proximal tubule is to reabsorb and process filtered proteins. Diabetes is characterized by increased quantities of tissue and circulating proteins modified by AGEs. Therefore in diabetes, plasma proteins filtered at the glomerulus and presented to the renal proximal tubule are likely to be highly modified by AGEs. METHODS: The model system was electrically resistant polarized renal proximal tubular epithelial cells in monolayer culture. The model proteins comprise a well-characterized AGE, methylglyoxal-modified bovine serum albumin (MGO-BSA), and unmodified BSA. RESULTS: Renal proximal tubular cells handle MGO-BSA and native BSA in markedly disparate ways, including differences in: (1) kinetics of binding, uptake, and intracellular accumulation, (2) processing and fragmentation, and (3) patterns of electrical conductance paralleling temporal changes in binding, uptake and processing. CONCLUSION: These differences support the idea that abnormal protein processing by the renal tubule can be caused by abnormal proteins, thereby forging a conceptual link between the pathogenic role of AGEs and early changes in tubular function that can lead to hypertrophy and nephropathy in diabetes.


Subject(s)
Diabetic Nephropathies/metabolism , Glycation End Products, Advanced/metabolism , Kidney Tubules, Proximal/metabolism , Pyruvaldehyde/pharmacokinetics , Serum Albumin, Bovine/pharmacokinetics , Cell Line, Transformed , Diabetic Nephropathies/pathology , Epithelial Cells/cytology , Epithelial Cells/metabolism , Fluorescein-5-isothiocyanate , Fluorescent Dyes , Humans , Kidney Tubules, Proximal/cytology , Microscopy, Confocal
2.
Ann N Y Acad Sci ; 1043: 625-36, 2005 Jun.
Article in English | MEDLINE | ID: mdl-16037286

ABSTRACT

Diabetes is characterized by increased quantities of circulating proteins modified by advanced glycation end products (AGEs). Proteins filtered at the glomerulus and presented to the renal proximal tubule are likely to be highly modified by AGEs. The proximal tubule binds, takes up, and catabolizes AGE-modified albumin by pathways different from those of unmodified albumin. These differences were examined in polarized, electrically resistant proximal tubular cells grown in monolayer culture. In patients with type 1 diabetes, urinary excretion of a lysosomal enzyme predicted the development of nephropathy.


Subject(s)
Diabetic Nephropathies/physiopathology , Glycation End Products, Advanced/metabolism , Kidney Tubules, Proximal/metabolism , Serum Albumin/metabolism , Acetylglucosaminidase/urine , Albuminuria/physiopathology , Cells, Cultured , Culture Media , Diabetes Mellitus, Type 1/physiopathology , Fluorescein-5-isothiocyanate/analogs & derivatives , Fluorescein-5-isothiocyanate/metabolism , Humans , Serum Albumin, Bovine/metabolism
3.
Am J Physiol Renal Physiol ; 289(2): F420-30, 2005 Aug.
Article in English | MEDLINE | ID: mdl-15814529

ABSTRACT

Chronic hyperglycemia causes structural alterations of proteins through the Maillard reaction. In diabetes, methylglyoxal (MGO)-induced hydroimidazolones are the predominant modification. In contrast to acute hyperglycemia, mitochondrial respiration is depressed in chronic diabetes. To determine whether MGO-derived protein modifications result in abnormalities in mitochondrial bioenergetics and superoxide formation, proteomics and functional studies were performed in renal cortical mitochondria isolated from rats with 2, 6, and 12 mo of streptozotocin-induced diabetes. MGO-modified proteins belonged to the following two pathways: 1) oxidative phosphorylation and 2) fatty acid beta-oxidation. Two of these proteins were identified as components of respiratory complex III, the major site of superoxide production in health and disease. Mitochondria from rats with diabetes exhibited a diminution of oxidative phosphorylation. A decrease in the respiratory complex III activity was significantly correlated with the quantity of MGO-derived hydroimidazolone present on mitochondrial proteins in both diabetic and control animals. In diabetes, isolated renal mitochondria produced significantly increased quantities of superoxide and showed evidence of oxidative damage. Administration of aminoguanidine improved mitochondrial respiration and complex III activity and decreased oxidative damage to mitochondrial proteins. Therefore, posttranslational modifications of mitochondrial proteins by MGO may represent pathogenic events leading to mitochondria-induced oxidative stress in the kidney in chronic diabetes.


Subject(s)
Diabetic Nephropathies/metabolism , Mitochondrial Proteins/metabolism , Oxidants/metabolism , Superoxides/metabolism , 2,4-Dinitrophenol/pharmacology , Animals , Blood Glucose/metabolism , Blotting, Western , Diabetes Mellitus, Experimental/metabolism , Electrophoresis, Gel, Two-Dimensional , Energy Metabolism , Fatty Acids/metabolism , Glucose/metabolism , Guanidines/pharmacology , Male , Mitochondria/drug effects , Mitochondria/metabolism , Mitochondrial Proteins/drug effects , Multienzyme Complexes/metabolism , NADH, NADPH Oxidoreductases/metabolism , Oxidation-Reduction , Oxidative Phosphorylation , Oxidative Stress/drug effects , Oxygen Consumption/drug effects , Pyruvaldehyde/pharmacology , Rats , Rats, Inbred Lew , Uncoupling Agents/pharmacology
4.
Oncogene ; 21(12): 1882-9, 2002 Mar 14.
Article in English | MEDLINE | ID: mdl-11896621

ABSTRACT

We recently identified inositol hexakisphosphate kinase 2 (IP6K2) as a positive regulator of apoptosis. Overexpression of IP6K2 enhances apoptosis induced by interferon-beta (IFN-beta) and cytotoxic agents in NIH-OVCAR-3 ovarian carcinoma cells. In this study, we contrast and compare IFN-beta and radiation-induced death, and show that IP6K2 expression sensitizes tumor cells. Unirradiated NIH-OVCAR-3 cells transfected with IP6K2 formed fewer colonies compared to unirradiated vector-expressing cells. IP6K2 overexpression caused increased radiosensitivity, evidenced by decreased colony forming units (CFU). Both IFN-beta and radiation induced caspase 8. IFN-beta, but not gamma-irradiation, induced TRAIL in NIH-OVCAR-3 cells. Gamma irradiation, but not IFN-beta, induced DR4 mRNA. Apoptotic effects of IFN-beta or gamma-irradiation were blocked by expression of a dominant negative mutant death receptor 5 (DR5Delta) or by Bcl-2. Caspase-8 mRNA induction was more pronounced in IP6K2-expressing cells compared to vector-expressing cells. These data suggest that overexpression of IP6K2 enhances sensitivity of some ovarian carcinomas to radiation and IFN-beta. IP6K2 may function to enhance the expression and/or function of caspase 8 and DR4 following cell injury. Both IFN-beta and gamma-irradiation induce apoptosis through the extrinsic, receptor-mediated pathway, IFN-beta through TRAIL, radiation through DR4, and both through caspase 8. The function of both death inducers is positively regulated by IP6K2.


Subject(s)
Ovarian Neoplasms/enzymology , Phosphotransferases (Phosphate Group Acceptor)/physiology , Antineoplastic Agents/therapeutic use , Apoptosis/drug effects , Apoptosis/radiation effects , Caspase 8 , Caspase 9 , Caspases/genetics , Caspases/metabolism , Female , Gamma Rays , Gene Expression Regulation, Enzymologic , Humans , Interferon-beta/pharmacology , Ovarian Neoplasms/drug therapy , Ovarian Neoplasms/radiotherapy , Proto-Oncogene Proteins c-bcl-2/physiology , RNA, Messenger/metabolism , Radiation-Sensitizing Agents , Receptors, TNF-Related Apoptosis-Inducing Ligand , Receptors, Tumor Necrosis Factor/genetics , Receptors, Tumor Necrosis Factor/metabolism , Receptors, Tumor Necrosis Factor/physiology , Ribonucleases/genetics , Ribonucleases/metabolism , Transfection , Tumor Cells, Cultured/drug effects , Tumor Cells, Cultured/radiation effects , Tumor Stem Cell Assay
SELECTION OF CITATIONS
SEARCH DETAIL
...