4-hydroxy-2-nonenal and Kidney-Diseases

The pathogenesis of crystal nephropathy involves deposition of intratubular crystals, tubular obstruction and cell death. The deposition of 8-dihydroxyadenine (DHA) crystals within kidney tubules, for instance, is caused by a hereditary deficiency of adenine phosphoribosyl transferase in humans or adenine overload in preclinical models. However, the downstream pathobiological patterns of tubular cell attrition in adenine/DHA-induced nephropathy remain poorly understood. In this study, we investigated: (i) the modes of adenine-induced tubular cell death in an experimental rat model and in human primary proximal tubular epithelial cells (PTEC); and (ii) the therapeutic effect of the flavonoid baicalein as a novel cell death inhibitor. In a rat model of adenine diet-induced crystal nephropathy, significantly elevated levels of tubular iron deposition and lipid peroxidation (4-hydroxynonenal; 4-HNE) were detected. This phenotype is indicative of ferroptosis, a novel form of regulated necrosis. In cultures of human primary PTEC, adenine overload-induced significantly increased mitochondrial superoxide levels, mitochondrial depolarisation, DNA damage and necrotic cell death compared with untreated PTEC. Molecular interrogation of adenine-stimulated PTEC revealed a significant reduction in the lipid repair enzyme glutathione peroxidase 4 (GPX4) and the significant increase in 4-HNE compared with untreated PTEC, supporting the concept of ferroptotic cell death. Moreover, baicalein treatment inhibited ferroptosis in adenine-stimulated PTEC by selectively modulating the mitochondrial antioxidant enzyme superoxide dismutase 2 (SOD2) and thus, suppressing mitochondrial superoxide production and DNA damage. These data identify ferroptosis as the primary pattern of PTEC necrosis in adenine-induced nephropathy and establish baicalein as a potential therapeutic tool for the clinical management of ferroptosis-associated crystal nephropathies (e.g., DHA nephropathy, oxalate nephropathy).

Topics: Adenine; Aldehydes; Animals; Epithelial Cells; Ferroptosis; Flavanones; Humans; Iron; Kidney Diseases; Kidney Tubules, Proximal; Lipid Peroxidation; Mitochondria; Rats; Superoxide Dismutase

Topics: Aldehydes; Animals; Cadmium; Cystathionine beta-Synthase; Cysteine; Environmental Pollutants; Glutathione; Glutathione Disulfide; Kidney; Kidney Diseases; Male; Malondialdehyde; Methionine Adenosyltransferase; Protective Agents; Pyrazines; Rats, Sprague-Dawley

Cisplatin (CP) is a potent and widely used chemotherapeutic agent. However, the clinical benefits of CP are compromised because it elicits nephrotoxicity and ototoxicity. In this study, we investigated the nephroprotective effects of the phytochemical genipin (GP) isolated from the gardenia (Gardenia jasminoides) fruit, using a murine model of CP-induced nephropathy. GP pretreatment attenuated the CP-induced renal tissue injury by diminishing the serum blood urea nitrogen, creatinine, and cystatin C levels, as well as those of kidney injury molecule-1. In addition, GP attenuated the CP-induced oxidative/nitrative stress by suppressing the activation of NADPH oxidase, augmenting the endogenous antioxidant defense system, and diminishing the accumulation of 4-hydroxynonenal and 3-nitrotyrosine in renal tissues. Furthermore, reduced levels of proinflammatory cytokines such as tumor necrosis factor-alpha and interleukin-1 beta indicated that CP-induced renal inflammation was mitigated upon the treatment with GP. GP also attenuated the CP-induced activation of mitogen-activated protein kinases, excessive activities of caspase-3/7 and poly(ADP-ribose) polymerase, DNA fragmentation, and apoptosis. When administered 12h after the onset of kidney injury, GP showed a therapeutic effect by ameliorating CP-induced nephrotoxicity. Moreover, GP synergistically enhanced the CP-induced cell death of T24 human bladder cancer cells. Collectively, our data indicate that GP attenuated the CP-induced renal tissue injury by abrogating oxidative/nitrative stress and inflammation and by blocking cell death pathways, thereby improving the renal function. Thus, our results suggest that the use of GP may be a promising new protective strategy against cisplatin-induced nephrotoxicity.

Topics: Aldehydes; Animals; Antioxidants; Apoptosis; Blood Urea Nitrogen; Caspase 3; Caspase 7; Cell Line, Tumor; Cisplatin; Creatinine; Cystatin C; Cytokines; Hepatitis A Virus Cellular Receptor 1; Humans; Inflammation; Iridoids; Kidney; Kidney Diseases; Male; Mice; Mice, Inbred C57BL; Mitogen-Activated Protein Kinases; Oxidative Stress; Poly(ADP-ribose) Polymerases; Tyrosine

NADPH oxidase Nox4-derived reactive oxygen species (ROS) play important roles in renal fibrosis. Our previous study demonstrated that intermedin (IMD) alleviated unilateral ureteral obstruction (UUO)-induced renal fibrosis by inhibition of ROS. However, the precise mechanisms remain unclear. Herein, we investigated the effect of IMD on Nox4 expression and NADPH oxidase activity in rat UUO model, and explored if these effect were achieved through cAMP-PKA pathway, the important post-receptor signal transduction pathway of IMD, in TGF-β1-stimulated rat proximal tubular cell (NRK-52E). Renal fibrosis was induced by UUO. NRK-52E was exposed to rhTGF-β1 to establish an in vitro model of fibrosis. IMD was overexpressed in the kidney and in NRK-52E by IMD gene transfer. We studied UUO-induced ROS by measuring dihydroethidium levels and lipid peroxidation end-product 4-hydroxynonenal expression. Nox4 expression in the obstructed kidney of UUO rat or in TGF-β1-stimulated NRK-52E was measured by quantitative RT-PCR and Western blotting. We analyzed NADPH oxidase activity using a lucigenin-enhanced chemiluminescence system. We showed that UUO-stimulated ROS production was remarkably attenuated by IMD gene transfer. IMD overexpression inhibited UUO-induced up-regulation of Nox4 and activation of NADPH oxidase. Consistent with in vivo results, TGF-β1-stimulated increase in Nox4 expression and NADPH oxidase activity was blocked by IMD. In NRK-52E, these beneficial effects of IMD were abolished by pretreatment with N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide hydrochloride (H-89), a PKA inhibitor, and mimicked by a cell-permeable cAMP analog dibutyl-cAMP. Our results indicate that IMD exerts anti-oxidant effects by inhibition of Nox4, and the effect can be mediated by cAMP-PKA pathway.

Topics: 8-Bromo Cyclic Adenosine Monophosphate; Adrenomedullin; Aldehydes; Animals; Cell Line; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Ethidium; Fibrosis; Gene Transfer Techniques; Isoquinolines; Kidney; Kidney Diseases; Lipid Peroxidation; Male; NADPH Oxidase 4; Neuropeptides; Oxidative Stress; Rats; Rats, Wistar; Reactive Oxygen Species; Real-Time Polymerase Chain Reaction; Signal Transduction; Sulfonamides; Transforming Growth Factor beta1; Up-Regulation

Activation of the farnesoid X receptor (FXR) has indicated a therapeutic potential for this nuclear bile acid receptor in the prevention of diabetic nephropathy and obesity-induced renal damage. Here, we investigated the protective role of FXR against kidney damage induced by obesity in mice that had undergone uninephrectomy, a model resembling the clinical situation of kidney donation by obese individuals. Mice fed a high-fat diet developed the core features of metabolic syndrome, with subsequent renal lipid accumulation and renal injury, including glomerulosclerosis, interstitial fibrosis, and albuminuria. The effects were accentuated by uninephrectomy. In human renal biopsies, staining of 4-hydroxynonenal (4-HNE), glucose-regulated protein 78 (Grp78), and C/EBP-homologous protein, markers of endoplasmic reticulum stress, was more prominent in the proximal tubules of 15 obese patients compared with 16 non-obese patients. In mice treated with the FXR agonist obeticholic acid, renal injury, renal lipid accumulation, apoptosis, and changes in lipid peroxidation were attenuated. Moreover, disturbed mitochondrial function was ameliorated and the mitochondrial respiratory chain recovered following obeticholic acid treatment. Culturing renal proximal tubular cells with free fatty acid and FXR agonists showed that FXR activation protected cells from free fatty acid-induced oxidative stress and endoplasmic reticulum stress, as denoted by a reduction in the level of reactive oxygen species staining and Grp78 immunostaining, respectively. Several genes involved in glutathione metabolism were induced by FXR activation in the remnant kidney, which was consistent with a decreased glutathione disulfide/glutathione ratio. In summary, FXR activation maintains endogenous glutathione homeostasis and protects the kidney in uninephrectomized mice from obesity-induced injury.

Topics: Aldehydes; Animals; Bile Acids and Salts; CCAAT-Enhancer-Binding Proteins; Chenodeoxycholic Acid; Diet, High-Fat; Endoplasmic Reticulum Chaperone BiP; Endoplasmic Reticulum Stress; Fatty Acids, Nonesterified; Female; Glutathione; Heat-Shock Proteins; Humans; Kidney; Kidney Diseases; Kidney Tubules; Lipid Metabolism; Lipid Peroxidation; Lipids; Male; Mice; Mice, Inbred C57BL; Mice, Obese; Middle Aged; Nephrectomy; Non-alcoholic Fatty Liver Disease; Obesity; Oxidative Stress; Reactive Oxygen Species; Receptors, Cytoplasmic and Nuclear

Dyslipidemia, particularly increased LDL-cholesterol level in serum, is associated with atherosclerosis and fibrosis in different organs. This study was designed to investigate the effects of increase in LDL-cholesterol on renal fibrosis.. Wild-type (WT) and LDLr knockout (KO) mice were fed standard or high fat diet (HFD), and their kidneys were collected after 26 weeks of dietary intervention for identification of fibrosis and study of potential mechanisms. Additional studies were performed in cultured renal fibroblasts.. We observed extensive and diffuse fibrosis in the kidneys of mice given HFD (P < 0.05 vs. standard chow). Fibrosis was associated with enhanced expression of fibronectin, nicotinamide adenine dinucleotide phosphate oxidases and activated p38 and p44/42 mitogen-activated protein kinases (MAPKs) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB). There was evidence for accumulation of 4-hydroxynonenal, a lipid peroxidation product, in the kidneys and of ox-LDL in the arteries of LDLr KO mice given HFD. The expression of ox-LDL receptor LOX-1 and of transforming growth factor beta 1 (TGFβ1) was increased in these kidneys. All these changes were more pronounced in LDLr KO mice than in the WT mice. In in vitro studies, treatment of fibroblasts from kidneys of LDLr KO mice with ox-LDL showed intense proliferation and collagen formation (all P < 0.05, fibroblasts from WT mice kidneys). Blockade of p38 MAPK, p44/42 MAPK, or NF-κB significantly attenuated expression of profibrotic signals, collagen formation, and proliferation of fibroblasts.. HFD induces renal fibrosis in LDLr-null mice primarily through activation of the nicotinamide adenine dinucleotide phosphate oxidase MAPK-NF-κB pathway by ox-LDL.

Topics: Aldehydes; Animals; Cell Proliferation; Cells, Cultured; Collagen; Diet, High-Fat; Fibroblasts; Fibrosis; Kidney Diseases; Lipid Peroxidation; Lipoproteins, LDL; Male; Mice; Mice, Knockout; Mitogen-Activated Protein Kinases; NADPH Oxidases; NF-kappa B; Receptors, LDL; Transforming Growth Factor beta1

Cisplatin, a cancer chemotherapy drug, is nephrotoxic. The aim of this study was to investigate whether resveratrol (RES) reduced cisplatin cytotoxicity and oxidative stress. Rat renal cortical slices were pre-incubated 30min with 0 (VEH, ethanol) or 30μg/ml RES followed by 60, 90 or 120min co-incubation with 0, 75, or 150μg/ml cisplatin. Lactate dehydrogenase (LDH) leakage was unchanged at 60 and 90min by cisplatin. Cisplatin increased (p<0.05) LDH leakage at 120min which was protected by RES. Cisplatin induced oxidative stress prior to LDH leakage as cisplatin depressed glutathione peroxidase and superoxide dismutase (SOD) activity, increased lipid peroxidation, protein carbonyls and 4-hydroxynonenal (4-HNE) adducted proteins within 60min. RES failed to reverse glutathione (GSH) depression by cisplatin. In order to eliminated an extracellular interaction between RES and cisplatin, additional studies (RINSE studies) allowed a 30min RES uptake into slices, transfer of slices to buffer lacking RES, followed by 120min cisplatin incubation. RES in the RINSE studies prevented LDH leakage by cisplatin indicating that RES protection was not via a physical interaction with cisplatin in the media. These findings indicate that RES diminished cisplatin in vitro renal toxicity and prevented the development of oxidative stress.

Topics: Aldehydes; Animals; Antineoplastic Agents; Antineoplastic Agents, Phytogenic; Antioxidants; Cell Survival; Cisplatin; Glutathione; Glutathione Peroxidase; Kidney; Kidney Diseases; L-Lactate Dehydrogenase; Lipid Peroxidation; Male; Oxidation-Reduction; Oxidative Stress; Protein Carbonylation; Rats; Rats, Inbred F344; Resveratrol; Stilbenes; Superoxide Dismutase; Tumor Necrosis Factor-alpha

Liver-type fatty acid-binding protein (L-FABP) in proximal tubules was reported to have renoprotective roles in experimental tubulointerstitial diseases via its anti-oxidative properties. Since tubuloglomerular cross-talk was recently discussed in the progression of renal diseases, to investigate whether tubular L-FABP may have an impact on the progression of glomerular damage, we induced IgA nephropathy (IgAN) in mice (Tg) transgenically tubular overexpressing human L-FABP (hL-FABP).. We reconstituted IgAN by bone marrow transplantation (BMT) from IgAN-prone mice into Tg and wild-type (WT) mice. Renal damage was evaluated at 6 and 12 weeks after BMT. During in vitro experiments, mesangial cells (MC) were stimulated by aggragated IgA (AIgA), and their supernatants (AIgA-MC medium) were collected. Stable cell line of mouse proximal tubular cell (mProx) transfected with or without hL-FABP gene was cultured with the AIgA-MC medium.. Although mesangial IgA deposition and serum IgA level were not different between WT (WT/ddY) and Tg (Tg/ddY) recipients, WT/ddY mice showed a significantly higher urinary albumin level and mesangial matrix expansion with a significantly higher glomerular damage score. Furthermore, CD68 + macrophage infiltration was also significantly attenuated in Tg/ddY mice. Up-regulation of renal hL-FABP was associated with significant suppression of renal heme oxygenase-1 (HO-1) expression and accumulation of 4-hydroxy-2-nonenal (4-HNE) and MCP-1 expression in Tg/ddY mice. In vitro experiments showed that AIgA-MC medium and recombinant TNF-α significantly up-regulated hL-FABP expression, which was partially blocked by anti-TNF-α antibody, and major mediators of oxidative stress (HO-1 and 4-HNE) and inflammation (MCP-1). Importantly, such up-regulation of the mediators in mProx with hL-FABP was significantly suppressed much more than that in mProx.. Tubular L-FABP activated by MC-origin humoral factors may lessen progression of glomerular damage at early stages of IgAN by reducing oxidative stress and inflammatory mediators.

Topics: Aldehydes; Animals; Blotting, Western; Cells, Cultured; Chemokine CCL2; Fatty Acid-Binding Proteins; Female; Glomerulonephritis, IGA; Heme Oxygenase-1; Humans; Inflammation; Kidney Diseases; Kidney Glomerulus; Kidney Tubules, Proximal; Mesoderm; Mice; Mice, Inbred C57BL; Mice, Transgenic; Oxidative Stress; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tumor Necrosis Factor-alpha

Cis-diamminedichloroplatinum II (CDDP)-induced nephrotoxicity is associated with the overproduction of reactive oxygen species. tert-Butylhydroquinone (tBHQ) is a compound widely used as food antioxidant. The purpose of this study was to investigate the ability of tBHQ to prevent the nephrotoxic effect of CDDP in rats as well as the mechanisms involved. Thirty-six Wistar rats divided in the following groups were used: control, tBHQ (12.5mg/kg), CDDP (7.5mg/kg) and tBHQ+CDDP. Twenty-four h urine was collected at the beginning and at the end of the experiment and the rats were sacrificed 72h after CDDP-administration. Histological studies were performed and markers of renal function and oxidative/nitrosative stress were measured. In addition, the activity of the following antioxidant enzymes was measured: glutathione peroxidase (GPx), superoxide dismutase (SOD), glutathione reductase (GR) and glutathione-S-transferase (GST). CDDP-induced renal dysfunction, structural damage and oxidative/nitrosative were prevented by tBHQ. In addition, tBHQ completely prevented the CDDP-induced fall in GPx and GST activities. In conclusion, the present study indicates that the antioxidant activity of tBHQ is associated with its nephroprotective effect against CDDP-induced acute kidney injury in rats.

Topics: Aldehydes; Animals; Antioxidants; Catalase; Cisplatin; Glutathione Peroxidase; Glutathione Reductase; Glutathione Transferase; Hydroquinones; Immunohistochemistry; Kidney Diseases; Male; Malondialdehyde; Oxidative Stress; Random Allocation; Rats; Rats, Wistar; Superoxide Dismutase

Cisplatin is a chemotherapeutic agent used in the treatment of several cancer tumors; however, nephrotoxicity has restricted its use. Reactive oxygen species and peroxynitrite, which is formed by the reaction between superoxide anion and nitric oxide (NO*), are implicated in cisplatin-induced nephrotoxicity. In contrast, both toxic and beneficial effects of NO* have been suggested in cisplatin-induced nephrotoxicity. Therefore, nowadays the role of NO* in this experimental model remains controversial. The aim of the present work was to elucidate the role of NO* in cisplatin-induced renal damage using N-[3-(aminomethyl)benzyl]acetamidine (1400W), a selective and irreversible inhibitor of iNOS. The mRNA levels of iNOS were increased in cisplatin-treated rats. The administration of 1400W reduced the cisplatin induced histological damage, renal dysfunction (increase in proteinuria and kidney injury molecule expression and decrease in creatinine clearance), tubulointerstitial infiltration, oxidative stress (increase in renal malondialdehyde and inmmunostaining for 4-hydroxy-2-nonenal) and nitrosative stress (immunostaining for 3-nitrotyrosine). In addition, the administration of 1400W was unable to modify systolic blood pressure in control rats. Our data demonstrate that selective iNOS inhibition reduces the cisplatin-induced nephrotoxicity and nitrosative stress which strongly suggest that in this experimental model (1) the NO* production is toxic and (2) iNOS is the main source of NO*.

Topics: Aldehydes; Amidines; Animals; Antineoplastic Agents; Benzylamines; Cisplatin; Disease Models, Animal; Enzyme Inhibitors; Gene Expression Regulation, Enzymologic; Kidney; Kidney Diseases; Male; Malondialdehyde; Nitric Oxide; Nitric Oxide Synthase Type II; Oxidative Stress; Rats; Rats, Wistar; RNA, Messenger; Tyrosine

The effect of the garlic-derived antioxidant S-allylcysteine (SAC) on renal injury and oxidative stress induced by ischemia and reperfusion (IR) was studied in this work. Rats were anesthetized and subjected to right nephrectomy; 15 min later ischemia was induced for a period of 40 min and then the rats were subjected to a reperfusion period of 6 h after which they were killed to obtain blood and the left kidney. SAC was given at a dose of 100 mg/kg 30 min before nephrectomy, 15 min before ischemia, immediately before reperfusion and 2 h after reperfusion. IR-induced renal injury was evident by the increase in blood urea nitrogen (BUN) and serum creatinine as well as by the renal structural damage which was assessed by histological analysis. IR-induced oxidative stress was evident by the increase in immunostaining with 4-hydroxy-2-nonenal (4-HNE). SAC treatment was able to ameliorate the increase in BUN and serum creatinine and to decrease the structural damage. This protective effect was associated with a decrease in the immunostaining for 4-HNE. It is concluded that the antioxidant properties of SAC are involved in its protective effect on renal ischemia and reperfusion injury.

Topics: Aldehydes; Animals; Antioxidants; Cysteine; Female; Garlic; Immunohistochemistry; Kidney; Kidney Diseases; Rats; Reperfusion Injury

The immunosuppressants ciclosporin (cyclosporin A, CsA) and tacrolimus can cause severe nephrotoxicity. Since CsA increases free radical formation, this study investigated whether an extract from Camellia sinensis, which contains several polyphenolic free radical scavengers, could prevent nephrotoxicity caused by CsA and tacrolimus. Rats were fed powdered diet containing polyphenolic extract (0-0.1%) starting 3 days before CsA or tacrolimus. Free radicals were trapped with alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone (POBN) and measured using an electron spin resonance spectrometer. Both CsA and tacrolimus decreased glomerular filtration rates (GFR) and caused tubular atrophy, vacuolization and calcification and arteriolar hyalinosis, effects that were blunted by treatment with dietary polyphenols. Moreover, CsA and tacrolimus increased POBN/radical adducts in urine nearly 3.5 fold. Hydroxyl radicals attack dimethyl sulfoxide (DMSO) to produce a methyl radical fragment. Administration of CsA or tacrolimus with (12)C-DMSO produced a 6-line spectrum, while CsA or tacrolimus given with (13)C-DMSO produced a 12-line ESR spectrum, confirming formation of hydroxyl radicals. 4-Hydroxynonenal (4-HNE), a product of lipid peroxidation, accumulated in proximal and distal tubules after CsA or tacrolimus treatment. ESR changes and 4-HNE formation were largely blocked by polyphenols. Taken together, these results demonstrate that both CsA and tacrolimus stimulate free radical production in the kidney, most likely in tubular cells, and that polyphenols minimize nephrotoxicity by scavenging free radicals.

Topics: Aldehydes; Animals; Camellia sinensis; Chromatography, High Pressure Liquid; Cyclosporine; Flavonoids; Free Radicals; Glomerular Filtration Rate; Glycerol; Immunohistochemistry; Immunosuppressive Agents; Kidney; Kidney Diseases; Male; Olive Oil; Phenols; Phytotherapy; Plant Oils; Polyphenols; Rats; Rats, Sprague-Dawley; Tacrolimus

Oxidative and nitrosative stress have been involved in gentamicin-induced nephrotoxicity. The purpose of this work was to study the effect of S-allylmercaptocysteine, a garlic derived compound, on gentamicin-induced oxidative and nitrosative stress and nephrotoxicity. In addition, the in vitro reactive oxygen species scavenging properties of S-allylmercaptocysteine were studied.. S-allylmercaptocysteine was able to scavenge hydroxyl radicals and singlet oxygen in vitro. In rats treated with gentamicin (70 mg/Kg body weight, subcutaneously, every 12 h, for 4 days), renal oxidative stress was made evident by the increase in protein carbonyl content and 4-hydroxy-2-nonenal, and the nitrosative stress was made evident by the increase in 3-nitrotyrosine. In addition, gentamicin-induced nephrotoxicity was evident by the: (1) decrease in creatinine clearance and in activity of circulating glutathione peroxidase, and (2) increase in urinary excretion of N-acetyl-beta-D-glucosaminidase, and (3) necrosis of proximal tubular cells. Gentamicin-induced oxidative and nitrosative stress and nephrotoxicity were attenuated by S-allylmercaptocysteine treatment (100 mg/Kg body weight, intragastrically, 24 h before the first dose of gentamicin and 50 mg/Kg body weight, intragastrically, every 12 h, for 4 days along gentamicin-treatment).. In conclusion, S-allylmercaptocysteine is able to scavenge hydroxyl radicals and singlet oxygen in vitro and to ameliorate the gentamicin-induced nephrotoxicity and oxidative and nitrosative stress in vivo.

Topics: Aldehydes; Animals; Biomarkers; Body Weight; Carboxylic Acids; Cysteine; Dinitrophenols; Energy Intake; Free Radical Scavengers; Gentamicins; Hydroxyl Radical; Immunohistochemistry; Kidney Cortex; Kidney Diseases; Kidney Glomerulus; Kidney Tubules, Proximal; Male; Oxidation-Reduction; Oxidative Stress; Proteins; Rats; Rats, Wistar; Reactive Oxygen Species; Tyrosine; Urine

We previously showed that mice lacking galectin-3/AGE-receptor 3 develop accelerated diabetic glomerulopathy. To further investigate the role of galectin-3/AGE-receptor function in the pathogenesis of diabetic renal disease, galectin-3 knockout (KO) and coeval wild-type (WT) mice were injected for 3 months with 30 microg/day of N(epsilon)-carboxymethyllysine (CML)-modified or unmodified mouse serum albumin (MSA). Despite receiving equal doses of CML, KO had higher circulating and renal AGE levels and showed more marked renal functional and structural changes than WT mice, with significantly higher proteinuria, albuminuria, glomerular, and mesangial area and glomerular sclerosis index. Renal 4-hydroxy-2-nonenal content and NFkappaB activation were also more pronounced in KO-CML vs. WT-CML. Kidney mRNA levels of fibronectin, laminin, collagen IV, and TGF-beta were up-regulated, whereas those of matrix metalloproteinase-2 and -14 were down-regulated, again more markedly in KO-CML than WT-CML mice. Basal and CML-induced RAGE and 80K-H mRNA levels were higher in KO vs. WT mice. MSA injection did not produce any significant effect in both genotypes. The association of galectin-3 ablation with enhanced susceptibility to AGE-induced renal disease, increased AGE levels and signaling, and altered AGE-receptor pattern indicates that galectin-3 is operating in vivo as an AGE receptor to afford protection toward AGE-dependent tissue injury.

Topics: Aldehydes; Animals; Cell Death; Cell Proliferation; Extracellular Matrix Proteins; Galectin 3; Glycation End Products, Advanced; Kidney; Kidney Cortex; Kidney Diseases; Kidney Glomerulus; Kinetics; Lysine; Matrix Metalloproteinases; Mice; Mice, Inbred C57BL; Mice, Knockout; NF-kappa B; Proteinuria; Receptor for Advanced Glycation End Products; Receptors, Immunologic; RNA, Messenger; Transforming Growth Factor beta

Ferric nitrilotriacetate (Fe-NTA) is a renal toxicant and carcinogen in rats and mice. We found that its administration results in formation of 4-hydroxy-2-nonenal (HNE) in the renal proximal tubule cells of rats, and 8-hydroxydeoxyguanosine (8-OHdG) adducts in their DNA, suggesting a role for oxidative stress. Since 2-mercaptoethane sulfonate (MESNA) and N-acetylcysteine (NAC), administered orally, have been shown to increase the kidney levels of free thiol groups, their influence on the renal toxicity and carcinogenicity induced by Fe-NTA was examined in the present study. Male Wistar rats were intraperitoneally injected with Fe-NTA (12 mg Fe/kg), and MESNA (100 mg/kg) or NAC (200 mg/kg) was given orally 1 h before and 1 h after this treatment. The animals were killed for tissue analyses 3 h after the Fe-NTA exposure. In accord with our previous reports, HNE-modified protein was detected in the proximal tubules of Fe-NTA-treated rats by means of immunohistochemistry. Likewise, levels of 8-OHdG in the renal nuclear DNA, lipid peroxides as thiobarbituric acid-reactive substances in the kidneys, and blood urea nitrogen and creatinine in the serum were significantly increased by the Fe-NTA treatment. All of these changes were completely inhibited by oral administration of MESNA or NAC. These results suggest that both of these compounds can prevent the oxidative stress induced by Fe-NTA.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Acetylcysteine; Aldehydes; Animals; Antioxidants; Carcinogens; Deoxyguanosine; DNA; DNA Damage; Ferric Compounds; Kidney; Kidney Diseases; Kidney Tubules, Proximal; Lipid Peroxidation; Male; Mesna; Nitrilotriacetic Acid; Oxidative Stress; Rats; Rats, Wistar; Thiobarbituric Acid Reactive Substances