Subjects with allergic reactions to drugs show in vivo polarized patterns of cytokine expression depending on the chronology of the clinical reaction

Manuscrito aceptado el 28 junio de 2000Background: The mechanisms involved in adverse drug reactions with an immunologic basis (ADRIB) can be antibody dependent, mainly IgE or T cell dependent (sensitized T cells). These mechanisms are regulated by a number of cytokines, including IL-2, IL-4, IL-5, IFN-γ, and TNF-α, which follow the classical TH1/TH2 immunologic paradigm. Although evidence for this has been seen in ex vivo studies, the results are heterogeneous, and few in vivo studies have been carried out in subjects with ADRIB. Objective: We studied a group of patients who experienced either immediate reactions (n = 10) or nonimmediate reactions (n = 9) to drugs to determine the cytokine pattern profile during the acute stage of the response, as well as after recovery. Methods: PBMCs were taken at different time intervals of 24 hours or less and 7, 15, and 30 days after the onset of the reaction, and the specific cytokine transcription and production were determined by using quantitative competitive RT-PCR and ELISA, respectively. Results: There was a transient polarized pattern corresponding to a TH1 response with IL-2, IFN-γ, and TNF-α in nonimmediate reactions and to a TH2 response with IL-4 in immediate reactions. Conclusions: This is the first in vivo demonstration of these TH1/TH2 patterns in subjects with ADRIB and confirms that çan immunologic process is occurring related to the mechanisms involved in the pathologic manifestation. These findings are relevant to the understanding of the pathophysiologic mechanisms involved in ADRIB, suggesting that further studies in this direction are warranted.This work was supported in part by CICYT SAF 96/0240, Consejería de Salud de la Junta de Andalucía, FIS 98/0861, and the Spanish Society for Allergy and Clinical Immunology

Cytoprotective–Antioxidant Effect of Brunfelsia grandiflora Extract on Neuron-like Cells

Brunfelsia grandiflora is a South American solanaceae widely used since long ago for its recognized medicinal properties. We have recently reported its chemical composition, showing a relevant number of bioactive compounds with antioxidant capacity, and proved the cytoprotective and antioxidative stress potential of B. grandiflora in cultured endothelial cells. Since B. grandiflora extracts have shown effects on the central nervous system, the present study was designed to show the potential cytoprotective capacity and the antioxidative stress potential of phenolic extracts from the plant on cultured neuron-like cells, as a model to reduce the presentation or effects of chronic diseases of the nervous system. To this end, we studied its reactive oxygen species (ROS)-reducing capacity, its antioxidant defense mechanisms, and some molecular markers involved in redox balance and apoptosis. The results show that cell survival and most changes in biomarkers related to oxidative status, ROS, reduced glutathione, glutathione peroxidase and reductase, malondialdehyde, and caspase 3/7 activity, and molecular expression of cell death-related genes (BAX, BNIP3, and APAF1), NFκB, SOD, and NRF2 (genes from oxidative stress—antioxidants) induced by oxidative stress were prevented by either co- or pretreatment of neuron-like cells with B. grandiflora extracts (25–200 µg/mL). The results demonstrate the chemoprotective potential of the plant and support its medicinal use

Cytoprotective–Antioxidant Effect of Brunfelsia grandiflora Extract on Neuron-like Cells

2022 Descuento MDPIBrunfelsia grandiflora is a South American solanaceae widely used since long ago for its recognized medicinal properties. We have recently reported its chemical composition, showing a relevant number of bioactive compounds with antioxidant capacity, and proved the cytoprotective and antioxidative stress potential of B. grandiflora in cultured endothelial cells. Since B. grandiflora extracts have shown effects on the central nervous system, the present study was designed to show the potential cytoprotective capacity and the antioxidative stress potential of phenolic extracts from the plant on cultured neuron-like cells, as a model to reduce the presentation or effects of chronic diseases of the nervous system. To this end, we studied its reactive oxygen species (ROS)-reducing capacity, its antioxidant defense mechanisms, and some molecular markers involved in redox balance and apoptosis. The results show that cell survival and most changes in biomarkers related to oxidative status, ROS, reduced glutathione, glutathione peroxidase and reductase, malondialdehyde, and caspase 3/7 activity, and molecular expression of cell death-related genes (BAX, BNIP3, and APAF1), NFκB, SOD, and NRF2 (genes from oxidative stress—antioxidants) induced by oxidative stress were prevented by either co- or pretreatment of neuron-like cells with B. grandiflora extracts (25–200 µg/mL). The results demonstrate the chemoprotective potential of the plant and support its medicinal use.Ministerio de Economía, Industria y Competitividad (España)Universidad Nacional Mayor de San MarcosSección Deptal. de Farmacología y Toxicología (Veterinaria)Depto. de Farmacología, Farmacognosia y BotánicaFac. de VeterinariaFac. de FarmaciaTRUEpubDescuento UC

Cytoprotective and Anti-Oxidative Stress Capacity of South American Medicinal Plant Brunfelsia grandiflora in Endothelial Cells

Author Contributions: JLR and LG designed the research study. JLR, LG, ZMC, MDLC and LIR performed the research. MRG, WBL, OP, MSFA, RM provided help and advice on cell culture. MRG analyzed the data. LG, JLR and WBL wrote the manuscript. All authors contributed to editorial changes in the manuscript. All authors read and approved the final manuscript. All authors have participated sufficiently in the work and agreed to be accountable for all aspects of the work.Background: Oxidative stress is a potential cause of cardiovascular pathologies, so the protection of endothelial cells and vascular tissues is essential to avoid such conditions, perhaps with the use of natural compounds rich in phenolic compounds with a proven high antioxidant capacity. The present study was designed to show the cytoprotective capacity of phenolic extracts from the plant Brunfelsia grandiflora, as well as describe its antioxidant defense mechanisms and the expression of some molecular markers involved in cellular protection. Methods: Human EA.hy926 cells were exposed to Brunfelsia grandiflora (B. grandiflora) extract (1, 10, 25, 50, 100, and 200 μg/mL) in co-treatment (22 h with 100 μM tert-Butyl hydroperoxide (t-BOOH) and B. grandiflora concentrations) and pre-treatment (18 h of B. grandiflora concentrations and then 200 μM t-BOOH for 4 h). Cell viability, reactive oxygen species (ROS) production, nitric oxide (NO) levels, caspase 3/7 activity, malondialdehyde (MDA) concentration, and reduction in reduced glutathione (GSH) levels, glutathione peroxidase (GPx), and glutathione reductase (GR) activity were measured, and real-time PCR molecular assays superoxide dismutase (SOD2), nuclear factor E2-related factor (NRF2), BCL-2-associated X protein (BAX), and B-cell lymphoma 2 (BCL2) were performed. Data were analyzed via one-way ANOVA followed by Tukey's post hoc test. Results: B. grandiflora bark extract, mainly at concentrations of 25, 50, 100, and 200 μg/mL, significantly (p < 0.05) reversed (pre-treatment and co-treatment) the deleterious effects of t-BOOH on EA.hy926 endothelial cells, which were significantly decreased cell viability, GSH activity, and SOD2 and NRF2 expression (p < 0.05); and significantly increased levels of ROS, NO, MDA, caspase-3/7 activity, GPx activity, GR activity, and the BAX/BCL2 ratio (p < 0.05). Conclusions: B. grandiflora extract was able to reduce the deleterious effects of t-BOOH on EA.hy926 endothelial cells, which may indicate its potential phytotherapeutic benefit against cytotoxic damage caused by chemical agents.Ministerio de Economía, Industria y Competitividad (España)Universidad Nacional de San MarcosDepto. de Farmacología, Farmacognosia y BotánicaSección Deptal. de Farmacología y Toxicología (Veterinaria)Fac. de FarmaciaTRUEpubAPC financiada por la UC

Oxidative Status before and after Renal Replacement Therapy: Differences between Conventional High Flux Hemodialysis and on-Line Hemodiafiltration

Hemodialysis patients experience high oxidative stress because of systemic inflammation and depletion of antioxidants. Little is known about the global oxidative status during dialysis or whether it is linked to the type of dialysis. We investigated the oxidative status before (pre-) and after (post-) one dialysis session in patients subjected to high-flux dialysis (HFD) or on-line hemodiafiltration (OL-HDF). We analyzed carbonyls, oxidized LDL (oxLDL), 8-hydroxy-2′-deoxyguanosine, and xanthine oxidase (XOD) activity as oxidative markers, and total antioxidant capacity (TAC), catalase, and superoxide dismutase activities as measures of antioxidant defense. Indices of oxidative damage (OxyScore) and antioxidant defense (AntioxyScore) were computed and combined into a global DialysisOxyScore. Both dialysis modalities cleared all markers (p < 0.01) except carbonyls, which were unchanged, and oxLDL, which increased post-dialysis (p < 0.01). OxyScore increased post-dialysis (p < 0.001), whereas AntioxyScore decreased (p < 0.001). XOD and catalase activities decreased post-dialysis after OL-HDF (p < 0.01), and catalase activity was higher after OL-HDF than after HFD (p < 0.05). TAC decreased in both dialysis modalities (p < 0.01), but remained higher in OL-HDF than in HFD post-dialysis (p < 0.05), resulting in a lower overall DialysisOxyScore (p < 0.05). Thus, patients on OL-HDF maintain higher levels of antioxidant defense, which might balance the elevated oxidative stress during dialysis, although further longitudinal studies are needed.Sin financiación4.546 JCR (2019) Q1, 17/89 Nutrition & Dietetics1.329 SJR (2019) Q1, 25/327 Food Science, 19/128 Nutrition and DieteticsNo data IDR 2019UE

Variations in Circulating Active MMP-9 Levels during Renal Replacement Therapy

Renal replacement therapy (RRT) is complicated by a chronic state of inflammation and a high mortality risk. However, different RRT modalities can have a selective impact on markers of inflammation and oxidative stress. We evaluated the levels of active matrix metalloproteinase (MMP)-9 in patients undergoing two types of dialysis (high-flux dialysis (HFD) and on-line hemodiafiltration (OL-HDF)) and in kidney transplantation (KT) recipients. Active MMP-9 was measured by zymography and ELISA before (pre-) and after (post-) one dialysis session, and at baseline and follow-up (7 and 14 days, and 1, 3, 6, and 12 months) after KT. Active MMP-9 decreased post-dialysis only in HFD patients, while the levels in OL-HDF patients were already lower before dialysis. Active MMP-9 increased at 7 and 14 days post-KT and was restored to baseline levels three months post-KT, coinciding with an improvement in renal function and plasma creatinine. Active MMP-9 correlated with pulse pressure as an indicator of arterial stiffness both in dialysis patients and KT recipients. In conclusion, active MMP-9 is better controlled in OL-HDF than in HFD and is restored to baseline levels along with stabilization of renal parameters after KT. Active MMP-9 might act as a biomarker of arterial stiffness in RRT.Sin financiación4.879 JCR (2020) Q2, 96/295 Biochemistry & Molecular Biology1.125 SJR (2020) Q2, 127/438 BiochemistryNo data IDR 2020UE

The anti-aging factor Klotho protects against acquired long QT syndrome induced by uremia and promoted by fibroblast growth factor 23

Background: Chronic kidney disease (CKD) is associated with increased propensity for arrhythmias. In this context, ventricular repolarization alterations have been shown to predispose to fatal arrhythmias and sudden cardiac death. Between mineral bone disturbances in CKD patients, increased fibroblast growth factor (FGF) 23 and decreased Klotho are emerging as important effectors of cardiovascular disease. However, the relationship between imbalanced FGF23-Klotho axis and the development of cardiac arrhythmias in CKD remains unknown. Methods: We carried out a translational approach to study the relationship between the FGF23-Klotho signaling axis and acquired long QT syndrome in CKD-associated uremia. FGF23 levels and cardiac repolarization dynamics were analyzed in patients with dialysis-dependent CKD and in uremic mouse models of 5/6 nephrectomy (Nfx) and Klotho deficiency (hypomorphism), which show very high systemic FGF23 levels. Results: Patients in the top quartile of FGF23 levels had a higher occurrence of very long QT intervals (> 490 ms) than peers in the lowest quartile. Experimentally, FGF23 induced QT prolongation in healthy mice. Similarly, alterations in cardiac repolarization and QT prolongation were observed in Nfx mice and in Klotho hypomorphic mice. QT prolongation in Nfx mice was explained by a significant decrease in the fast transient outward potassium (K+) current (Itof), caused by the downregulation of K+ channel 4.2 subunit (Kv4.2) expression. Kv4.2 expression was also significantly reduced in ventricular cardiomyocytes exposed to FGF23. Enhancing Klotho availability prevented both long QT prolongation and reduced Itof current. Likewise, administration of recombinant Klotho blocked the downregulation of Kv4.2 expression in Nfx mice and in FGF23-exposed cardiomyocytes. Conclusion: The FGF23-Klotho axis emerges as a new therapeutic target to prevent acquired long QT syndrome in uremia by minimizing the predisposition to potentially fatal ventricular arrhythmias and sudden cardiac death in patients with CKD.Instituto de Salud Carlos III, Ministry of Economy, Industry and Competitiveness (PI17/01093, PI17/01193, PI20/00763, CP15/00129, F18/00261, CPII20/00022, SAF2017-84777-R, PID2020-113238RB-I00)Sociedad Española de Cardiología (SEC), and Fundación Renal Íñigo Álvarez de Toledo (FRIAT), co-funded by the European Regional Development Fund (Fondos FEDER)8.775 JCR (2020) Q1, 13/167 Medicine, General & Internal3.463 SJR (2020) Q1, 61/2446 Medicine (miscellaneous)No data IDR 2020UE

Variations in Circulating Active MMP-9 Levels during Renal Replacement Therapy

Renal replacement therapy (RRT) is complicated by a chronic state of inflammation and a high mortality risk. However, different RRT modalities can have a selective impact on markers of inflammation and oxidative stress. We evaluated the levels of active matrix metalloproteinase (MMP)-9 in patients undergoing two types of dialysis (high-flux dialysis (HFD) and on-line hemodiafiltration (OL-HDF)) and in kidney transplantation (KT) recipients. Active MMP-9 was measured by zymography and ELISA before (pre-) and after (post-) one dialysis session, and at baseline and follow-up (7 and 14 days, and 1, 3, 6, and 12 months) after KT. Active MMP-9 decreased post-dialysis only in HFD patients, while the levels in OL-HDF patients were already lower before dialysis. Active MMP-9 increased at 7 and 14 days post-KT and was restored to baseline levels three months post-KT, coinciding with an improvement in renal function and plasma creatinine. Active MMP-9 correlated with pulse pressure as an indicator of arterial stiffness both in dialysis patients and KT recipients. In conclusion, active MMP-9 is better controlled in OL-HDF than in HFD and is restored to baseline levels along with stabilization of renal parameters after KT. Active MMP-9 might act as a biomarker of arterial stiffness in RRT