8-hydroxy-2--deoxyguanosine and Hypertrophy

Expression of klotho, the renoprotective anti-aging gene, is decreased in diabetic model kidneys. We hypothesized that klotho protein attenuates renal hypertrophy and glomerular injury in a mouse model of diabetic nephropathy.. Klotho transgenic (KLTG) mice were crossed with spontaneously diabetic Ins2Akita (AKITA) mice. Glomerular morphology, macrophage infiltration, urinary albumin excretion and urinary 8-hydroxy-2-deoxy guanosine excretion were examined. In vitro, human glomerular endothelial cells were stimulated with high glucose with or without recombinant klotho, and calpain activity and proinflammatory cytokine expressions were measured.. We found that klotho protein overexpression attenuates renal hypertrophy and glomerular injury in this mouse model of diabetic nephropathy. Klotho overexpression attenuated renal hypertrophy, albuminuria, glomerular mesangial expansion, and endothelial glycocalyx loss in the AKITA mice. AKITA mice exhibit high levels of urinary 8-hydroxy-2-deoxy guanosine excretion. In the presence of klotho overexpression, this effect was reversed. In addition, the glomerular macrophage infiltration characteristic of AKITA mice was attenuated in KLTG-AKITA mice. In human glomerular endothelial cells, high glucose induced calpain activity. This effect was suppressed by expression of recombinant klotho, which also suppressed the induction of proinflammatory cytokines.. Our data suggest klotho protein protects against diabetic nephropathy through multiple pathways.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Albuminuria; Animals; Biomarkers; Calpain; Cells, Cultured; Deoxyguanosine; Diabetes Mellitus; Diabetic Nephropathies; Disease Models, Animal; Genotype; Glucose; Glucuronidase; Humans; Hypertrophy; Inflammation Mediators; Kidney Glomerulus; Klotho Proteins; Macrophages; Mice, Inbred C57BL; Mice, Transgenic; Phenotype; Transfection

The objective of this prospective, controlled study was to evaluate oxidative DNA damage in children with obstructive adenotonsillar hypertrophy. This study included 30 patients with obstructive adenotonsillar hypertrophy (male/female ratio, 3:2; age range, 3-9 y) scheduled to undergo tonsillectomy and adenoidectomy and 25 control subjects of similar age and sex with no adenotonsillar disease or airway obstruction. Urine and blood samples were obtained from each child for 8-hydroxy 2-deoxyguanosine (8-OhdG) and malondialdehyde (MDA) concentrations. There were significant differences in leukocyte (3.28 [0.69/10] vs 0.70 [0.15/10] dG) and urine 8-OhdG (8.22 [2.27/10] vs 5.26 [1.3/10] dG) levels in patients with obstructive adenotonsillar hypertrophy and healthy subjects (P < 0.001 for both). Plasma (2.98 [1.31] vs 1.14 [0.64] μM) and urine (1.77 [0.84] vs 0.56 [0.32] μM) MDA levels were also different (P < 0.001 for both). There were positive correlations between 8-OhdG in leukocyte DNA and plasma MDA (r = 0.648, P < 0.001) and between levels of urine 8-OhdG excretion and urine MDA (r = 0.588, P < 0.001). The DNA damage in children with adenotonsillar hypertrophy should be kept in mind, but further studies must be done with larger patient groups.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Adenoidectomy; Adenoids; Airway Obstruction; Case-Control Studies; Child; Child, Preschool; Deoxyguanosine; DNA Damage; Female; Humans; Hypertrophy; Leukocytes; Male; Malondialdehyde; Oxidative Stress; Palatine Tonsil; Prospective Studies; Tonsillectomy

Oxidative stress has been suggested to play an important role in the pathogenesis of diabetic nephropathy because it increases under diabetic conditions and is known to induce cellular dysfunction in a wide variety of cells. To protect cells against oxidative stress, cells possess defensive mechanisms such as intracellular antioxidants. Although it has been reported that central enzymes in the antioxidative defense mechanisms of the cell are induced under hyperglycemic conditions, the oxidative stress level remains high. On the other hand, there are endogenous inhibitors of antioxidants, such as thioredoxin interacting protein (Txnip). In the present study, the relationship between diabetic nephropathy and Txnip was investigated using streptozotocin (STZ)-induced diabetic mice. Eight-week-old male C57BL/6 mice were treated with either STZ or citrate vehicle. After 24 weeks of treatment, diabetic nephropathy and oxidative stress were assessed by biochemical analyses of urine and histological analyses of the kidneys. In addition, the expression of Type IV collagen alpha1 chain (Col4A1), Transforming growth factor-beta (TGF-beta), and Txnip were evaluated by real-time polymerase chain reaction. Albuminuria, renal hypertrophy, and expansion of the mesangial area, which are the hallmarks of diabetic nephropathy, were confirmed in the diabetic mice. The mRNA expression of COL4A1 and TGF-beta was dramatically increased in diabetic mice in comparison with the control mice. Moreover, associated with the increased renal expression of Txnip, diabetic conditions increased oxidative stress as determined by urinary excretion of 8-hydroxy-2'-deoxyguanosine and acrolein adduct, which are oxidative stress markers. Moreover, Txnip may be a therapeutic target in diabetic nephropathy.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Acrolein; Albuminuria; Animals; Biomarkers; Carrier Proteins; Collagen Type IV; Deoxyguanosine; Diabetic Nephropathies; Hypertrophy; Kidney; Male; Mice; Mice, Inbred C57BL; Oxidative Stress; RNA, Messenger; Streptozocin; Thioredoxins; Transforming Growth Factor beta

External esophageal perfusion (EEP) with the idea that esophageal perfusion can be controlled with a single ingredient at a constant rate and concentration, might be used to dissect the injurious role of gastro-duodenal secretions for the progression from esophagitis to Barrett's esophagus (BE) and esophageal adenocarcinoma (EAC). This study is to evaluate the EEP rat model for esophagitis induced by using a micro-osmotic pump with bile perfusion.. Eighteen adult rats underwent the EEP procedure. Bile (0.5% bovine bile, pH 7.4) was used as perfusion agent and three types of perfusions were performed: 1 week perfusion, 2 weeks perfusion, and 4 weeks perfusion compared to saline perfusion and sham operation. Histological changes, cell proliferation, apoptosis, 8-hydroxy-deoxyguanosine (8-OH-dG) and Manganese superoxide dismutase (MnSOD) were observed after perfusion and compared.. The bile perfusion for 1 week, 2 weeks, and 4 weeks induced mucosa infiltration of inflammatory cells, basal cell hyperproliferation, and papillae hypertrophy in all animals. Histopathology and cellular changes consistent with the findings associated with reflux esophagitis. The apoptotic index, the proliferating index, and expression of 8-OH-dG were significantly increased in the esophageal mucosa compared to controls. MnSOD expression was decreased with bile perfusion compared to saline controls.. The external esophageal perfusion model enabled precise control of the injurious agent. It induced the typical histological injury and cellular changes seen in severe reflux esophagitis. The cellular changes in apoptosis, proliferation and anti-oxidant defense make this model unique for reflux esophagitis studies. Further studies are needed to induce Barrett's esophagus and esophageal adenocarcinoma.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Animals; Apoptosis; Bile; Cell Division; Deoxyguanosine; Esophagitis, Peptic; Esophagus; Gastric Juice; Hypertrophy; Immunohistochemistry; In Situ Nick-End Labeling; Mucous Membrane; Perfusion; Rats; Rats, Sprague-Dawley; Superoxide Dismutase; Time Factors

An important role of redox regulation in myocardial diseases and heart failure has been postulated. Thioredoxin (TRX) is a redox-regulating protein. Recent studies indicated a possible association between plasma TRX concentrations and the severity of heart failure. Accordingly, we investigated the myocardial expression of TRX in patients with myocarditis and cardiomyopathies. Four cases of hypertrophic cardiomyopathy (HCM), 10 of dilated cardiomyopathy (DCM), 6 of myocarditis, and 5 of controls were studied. Right and left ventricular endomyocardial biopsy samples were obtained at the diagnostic cardiac catheterization. The samples were processed for immunohistological staining for TRX, which was done by the indirect immunoperoxidase technique. 8-hydoxy-2'-deoxyguanosine (8-OHdG), one of the major DNA base-modified products, was also detected for an established marker for oxidative stress. TRX immunoreactivity was none or trivial in control specimens. Positive TRX staining was found in 6 cases; 3 in active myocarditis and 3 in DCM. The positive staining was found in infiltrating cells and damaged myocytes in the perinecrotic lesions. Damaged myocytes were also positive for 8-OHdG All the 3 cases of DCM positive for TRX stain showed severe left ventricular hypertrophy on electrocardiogram and highly elevated left ventricular end-diastolic pressure (> 24 mmHg), suggesting the overload of oxidative stress by hemodynamic impairment. Myocardial TRX was upregulated in myocarditis and cardiomyopathies with active necrotic stage associated with DNA damage, which may reflect the oxidative stress overload in hemodynamically uncontrolled status.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Biopsy; Cardiomyopathies; Deoxyguanosine; Diastole; DNA; DNA Damage; Heart Ventricles; Humans; Hypertrophy; Immunohistochemistry; Myocarditis; Myocardium; Necrosis; Oxidation-Reduction; Oxidative Stress; Pressure; Thioredoxins; Up-Regulation