Cyclosporine A-induced apoptosis in renal tubular cells is related to oxidative damage and mitochondrial fission.

Cyclosporine A (CsA) nephrotoxicity has been linked to reactive oxygen species (ROS) production in renal cells. We have demonstrated that the antioxidant Vitamin E (Vit E) abolished renal toxicity in vivo and in vitro models. As one of the main sources of intracellular ROS are mitochondria, we studied the effects of CsA on several mitochondrial functions in LLC-PK1 cells. CsA induced ROS synthesis and decreased reduced glutathione (GSH). The drug decreased mitochondrial membrane potential (ΔΨm) and induced physiological modifications in both the inner (IMM) and the outer mitochondrial membranes (OMM). In the IMM, CsA provoked mitochondrial permeability transition pores (MPTP) and cytochrome c was liberated into the intermembrane space. CsA also induced pore formation in the OMM, allowing that intermembrane space contents can reach cytosol. Furthermore, CsA altered the mitochondrial dynamics, inducing an increase in mitochondrial fission; CsA increased the expression of dynamin related protein 1 (Drp1) that contributes to mitochondrial fission, and decreased the expression of mitofusin 2 (Mfn2) and optic atrophy protein 1 (Opa1), proteins involved in the fusion process. All these phenomena were related to apoptosis. These effects were inhibited when cells were treated with the antioxidant Vit E suggesting that they were mediated by the synthesis of ROS.

The role of CCR5/CXCR3 expressing CD8+ cells in liver damage and viral control during persistent hepatitis C virus infection.

BACKGROUND/AIMS: CXCR3 and CCR5 play a major role in recruiting cytotoxic T cells (Tc) and secreting secondary type 1 cytokines (Tc1) in the liver. HCV could impair their expression as a survival mechanism. The role of these chemokine receptors on CD8+ cells in chronic hepatitis C is analysed. METHODS: Serum, chemokines, peripheral blood and intrahepatic lymphocytes from chronic hepatitis C patients were studied. CXCR3/CCR5 expressing CD8+ cells were quantified by flow-cytometry. Serum chemokines concentration (CXCL10/CCL3) was measured by ELISA. Basal data were correlated with liver inflammation. Longitudinal data were obtained during treatment and correlated with virologic response. RESULTS: CCR5/CXCR3 expressing CD8+ cells were enriched in the liver and correlated with inflammation. Chronic HCV patients presented the same frequency of CCR5(high)/CXCR3(high) expressing CD8+ cells in peripheral blood as in healthy controls but higher serum concentration of CXCL10/CCL3. Treatment with PEG-interferon alpha-2b plus ribavirin increased CCR5(high)/CXCR3(high) expressing CD8+ cells frequency in peripheral blood and decreased CXCL10/CCL3 serum concentration. Increase in CXCR3(high) expressing CD8+ cells after 24 weeks of treatment was correlated with SVR. CONCLUSIONS: In chronic hepatitis C, anti-viral treatment induces an increase in CD8+ cells expressing chemokine receptors associated with Tc1 response and a reduction in their ligands. Achievement of viral control is associated with an increase in CXCR3(high) expressing CD8+ cells during treatment.

Cyclooxygenase-independent inhibition of H2O2-induced cell death by S-ketoprofen in renal cells.

The stress response of the distal tubule to oxidative attack may be relevant to recovery from acute renal failure. In distal tubular Madin-Darby cells (MDCK), H(2)O(2) induced up-regulation of cyclooxygenases (COX-1 and COX-2), prostaglandin-E(2) production and caspase-independent cell death. Cell death was inhibited by S-ketoprofen, but not by the much weaker COX inhibitor R-ketoprofen. Interestingly, we identified 15-deoxy-Delta(12,14)-prostaglandin-J(2) (15d-PGJ(2)), a peroxisome-proliferator activated receptor-gamma agonist, as a lethal prostaglandin whose effect was reproduced by the PPAR-gamma agonist ciglitazone. Nevertheless, H(2)O(2)-induced cell death was unaffected by other non-steroidal anti-inflammatory drugs (NSAIDs) or all-trans-retinoic acid. Moreover, c-Jun-N-terminal kinase inhibitor SP600125 prevented 15-deoxy-Delta(12,14)-PGJ(2)-induced cell death, but not H(2)O(2)-induced cell death. PPAR-gamma antagonist GW9662 showed no affect on the cell death. These results indicated that protection by S-ketoprofen was COX-independent and PPARgamma independent. Moreover, the IC(50) value of the action of S-ketoprofen for the inhibition of H(2)O(2)-induced MDCK cell death ( approximately equal 140microM) was much higher than the IC(50) value for the inhibition of COX-1 and COX-2 activities ( approximately equal 1microM). Further design of S-ketoprofen derivatives devoid of COX inhibitory activity will give opportunity to protect the kidney against oxidative attack while avoiding unwanted effects of NSAID.

[Systemic and lung inflammation in 2 phenotypes of chronic obstructive pulmonary disease]. / Inflamación pulmonar y sistémica en 2 fenotipos de EPOC.

OBJECTIVE: To study whether patients with chronic obstructive pulmonary disease (COPD) at the same level of flow limitation but with different clinical phenotypes present different degrees of systemic and/or pulmonary inflammation. PATIENTS AND METHODS: We studied 15 male smokers without COPD (control group) and 39 males with COPD in stable clinical condition. The COPD patients were assigned to 2 groups based on the ratio of carbon monoxide diffusing capacity (DLCO) to alveolar volume (DLCO/VA) expressed as a percentage as follows: a) mainly emphysema (n = 15) and b) mainly chronic bronchitis (n = 24). Classification was determined by comparing both clinical features and diagnostic images. RESULTS: Mean (SD) concentrations of interleukin 8 (IL-8) and 8-isoprostane in exhaled breath condensate (EBC) were significantly lower in patients with mainly emphysema (IL-8, 0.34 [0.70] pg/mL; 8-isoprostane, 0.07 [0.26] pg/mL) than in patients with chronic bronchitis (IL-8, 2.32 [3.10] pg/mL; 8-isoprostane, 1.77 [2.98] pg/mL) or in the controls (IL-8, 3.14 [4.59] pg/mL; 8-isoprostane, 1.92 [2.84] pg/mL); P < .05 for IL-8 comparisons and P < .01 for 8-isoprostane. IL-8, leukotriene B4, and 8-isoprostano in EBC correlated significantly with DLCO/VA (% of predicted) (r = 0.30, P < .05; r = 0.29, P < or = .05; and r = 0.46, P < .01, respectively) but not with forced expiratory volume in 1 second. There was a negative correlation between EBC and serum levels of both IL-8 (r = -0.31; P < .05) and 8-isoprostane (r = -0.51; P < .001). The correlation between leukotriene B4 concentrations in EBC and serum was not significant, however. No significant differences were found between smokers' and ex-smokers' serum levels of IL-8, leukotriene B4, 8-isoprostane in serum or EBC. CONCLUSIONS: The results indicate that COPD patients with an emphysematous phenotype have a less intense inflammatory response and less oxidative stress in the lung.

Inflamación pulmonar y sistémica en 2 fenotipos de EPOC / Systemic and lung inflammation in 2 phenotypes of Chronic Obstructive Pulmonary Disease

Objetivo: Investigar si los pacientes con enfermedad pulmonar obstructiva crónica (EPOC) con un mismo grado de limitación ventilatoria, pero diferente fenotipo clínico, presentan diferencias en el grado de respuesta inflamatoria pulmonar y/o sistémica. Pacientes y métodos: Se estudió a 15 varones fumadores sin EPOC (grupo control) y a 39 varones con EPOC en situación clínica estable. Usando la relación factor de transferencia de monóxido de carbono/volumen alveolar (TLCO/VA%), se dividió a los pacientes con EPOC en 2 grupos: a) EPOC de predominio enfisema (EPOC-A; n = 15), y b) EPOC de predominio bronquitis crónica (EPOC-B; n = 24). La correcta clasificación de los pacientes se confirmó analizando aspectos clínicos y técnicas de imagen. Resultados: Las concentraciones medias ± DE de interleucina-8 (IL-8) y de 8-isoprostano en el condensado de aire exhalado (CAE) fueron significativamente menores (p < 0,05 para la IL-8 y p < 0,01 para el 8-isoprostano) en los pacientes con predominio enfisematoso (IL-8: 0,34 ± 0,70 pg/ml; 8-isoprostano: 0,07 ± 0,26 pg/ml) que en los pacientes con bronquitis crónica (IL-8: 2,32 ± 3,10 pg/ml; 8-isoprostano: 1,77 ± 2,98 pg/ml) o que en los controles (IL-8: 3,14 ± 4,59 pg/ml; 8-isoprostano: 1,92 ± 2,84 pg/ml). Los valores de IL-8, leucotrieno B4 y 8-isoprostano en el CAE se relacionaron significativamente con los valores de TLCO/VA% (r = 0,30, p < 0,05; r = 0,29, p ≤ 0,05, y r = 0,46; p < 0,01, respectivamente), pero no con el volumen espiratorio forzado en el primer segundo. Existió una relación negativa entre los valores de IL-8 (r = -­0,31; p < 0,05) y 8-isoprostano (r = ­-0,51; p < 0,001) en suero y CAE. Sin embargo, esta correlación no fue significativa para el leucotrieno B4. No se observaron diferencias significativas entre fumadores activos y ex fumadores para IL-8, leucotrieno B4 y 8-isoprostano en suero y CAE. Conclusiones: Los resultados de este estudio indican que en pacientes con EPOC la presencia de un fenotipo enfisematoso se acompaña de una menor respuesta inflamatoria y menor estrés oxidativo en el pulmón

Objective: To study whether patients with chronic obstructive pulmonary disease (COPD) at the same level of flow limitation but with different clinical phenotypes present different degrees of systemic and/or pulmonary inflammation. Patients and methods: We studied 15 male smokers without COPD (control group) and 39 males with COPD in stable clinical condition. The COPD patients were assigned to 2 groups based on the ratio of carbon monoxide diffusing capacity (DLCO) to alveolar volume (DLCO/VA) expressed as a percentage as follows: a) mainly emphysema (n=15) and b) mainly chronic bronchitis (n=24). Classification was determined by comparing both clinical features and diagnostic images. Results: Mean (SD) concentrations of interleukin 8 (IL-8) and 8-isoprostane in exhaled breath condensate (EBC) were significantly lower in patients with mainly emphysema (IL-8, 0.34 [0.70] pg/mL; 8-isoprostane, 0.07 [0.26] pg/mL) than in patients with chronic bronchitis (IL-8, 2.32 [3.10] pg/mL; 8-isoprostane, 1.77 [2.98] pg/mL) or in the controls (IL-8, 3.14 [4.59] pg/mL; 8-isoprostane, 1.92 [2.84] pg/mL); P<.05 for IL-8 comparisons and P<.01 for 8-isoprostane. IL-8, leukotriene B4, and 8-isoprostano in EBC correlated significantly with DLCO/VA (% of predicted) (r=0.30, P<.05; r=0.29, P≤.05; and r=0.46, P<.01, respectively) but not with forced expiratory volume in 1 second. There was a negative correlation between EBC and serum levels of both IL-8 (r=-­0.31; P<.05) and 8-isoprostane (r=-­0.51; P<.001). The correlation between leukotriene B4 concentrations in EBC and serum was not significant, however. No significant differences were found between smokers' and ex-smokers' serum levels of IL-8, leukotriene B4, 8-isoprostane in serum or EBC. Conclusions: The results indicate that COPD patients with an emphysematous phenotype have a less intense inflammatory response and less oxidative stress in the lung