Presence Of Diclofenac Research Articles

Naproxen affects multiple organs in fish but is still an environmentally better alternative to diclofenac

The presence of diclofenac in the aquatic environment and the risks for aquatic wildlife, especially fish, have been raised in several studies. One way to manage risks without enforcing improved wastewater treatment would be to substitute diclofenac (when suitable from a clinical perspective) with another non-steroidal anti-inflammatory drug (NSAID) associated with less environmental risk. While there are many ecotoxicity-studies of different NSAIDs, they vary extensively in set-up, species studied, endpoints and reporting format, making direct comparisons difficult. We previously published a comprehensive study on the effects of diclofenac in the three-spined stickleback (Gasterosteus aculeatus). Our present aim was to generate relevant effect data for another NSAID (naproxen) using a very similar setup, which also allowed direct comparisons with diclofenac regarding hazards and risks. Sticklebacks were therefore exposed to naproxen in flow-through systems for 27 days. Triplicate aquaria with 20 fish per aquarium were used for each concentration (0, 18, 70, 299 or 1232 μg/L). We investigated bioconcentration, hepatic gene expression, jaw lesions, kidney and liver histology. On day 21, mortalities in the highest exposure concentration group unexpectedly reached ≥ 25 % in all three replicate aquaria, leading us to terminate and sample that group the same day. On the last day (day 27), the mortality was also significantly increased in the second highest exposure concentration group. Increased renal hematopoietic hyperplasia was observed in fish exposed to 299 and 1232 μg/L. This represents considerably higher concentrations than those expected in surface waters as a result of naproxen use. Such effects were observed already at 4.6 μg/L in the experiment with diclofenac (lowest tested concentration). Similar to the responses to diclofenac, a concentration-dependent increase in both relative hepatic gene expression of c7 (complement component 7) and jaw lesions were observed, again at concentrations considerably higher than expected in surface waters. Naproxen bioconcentrated less than diclofenac, in line with the observed effect data. An analysis of recent sales data and reported concentrations in treated sewage effluent in Sweden suggest that despite higher dosages used for naproxen, a complete substitution would only be expected to double naproxen emissions. In summary, naproxen and diclofenac produce highly similar effects in fish but the environmental hazards and risks are clearly lower for naproxen. Hence, if there are concerns for environmental risks to fish with diclofenac, a substitution would be advisable when naproxen presents an adequate alternative from a clinical point-of-view.

Diclofenac exhibits synergism with azoles against planktonic cells and biofilms of Candida tropicalis

This study aimed to evaluate the effect of diclofenac on minimum inhibitory concentrations of antifungals against planktonic cells and biofilms of Candida tropicalis. Susceptibility testing of planktonic cells was evaluated using the broth microdilution assay and checkerboard method. Biofilm formation by C. tropicalis in the presence of diclofenac, alone or in combination with antifungals, was also evaluated, and scanning electron microscope (SEM) and confocal microscope (CLSM) analyses were performed. Diclofenac showed an MIC of 1024 μg ml−1 against planktonic cells. The MICs of fluconazole and voriconazole against azole-resistant isolates were reduced 8- to 32-fold and 16- to 256-fold, respectively, when in combination with diclofenac. When in combination with fluconazole or voriconazole, diclofenac reduced the antifungal concentration necessary to inhibit C. tropicalis biofilm formation. In conclusion, diclofenac presents synergism with fluconazole and voriconazole against resistant C. tropicalis strains and improves the activity of these azole drugs against biofilm formation.

Acetoacetate enhances oxidative metabolism and response to toxicants of cultured kidney cells

Cultured kidney cells maintained in conventional growth media with high glucose levels exhibit increased glycolytic activity compared to the cells in vivo. In contrast, renal proximal tubules utilize substrates such as ketone bodies and rely on mitochondrial oxidative phosphorylation. LLC-PK1 cells maintain many features of the proximal tubule but are exposed to glucose concentrations ranging from 17 to 25 mM. This may impact their reliability in predicting mitochondrial toxicity. This study is designed to test the impact of the ketone body acetoacetate on metabolic characteristics of LLC-PK1 cells. Basal respiration, maximal respiration, spare respiratory capacity and ATP-linked respiration were significantly increased in cells grown in growth medium supplemented with 5 mM acetoacetate. In contrast, glycolytic capacity, as well as glycolytic reserve were significantly reduced in the acetoacetate group. There was an increased expression in biomarkers of mitochondrial biogenesis, and an increase in mitochondrial protein expression. Cells grown in medium complemented with acetoacetate displayed a significantly lower LC50 when treated with clotrimazole and diclofenac. There was a marked increase in uncoupled respiration in the presence of diclofenac, while clotrimazole and ciprofibrate significantly decreased respiration in the acetoacetate. The results indicate that acetoacetate complemented media can alter cellular metabolism and increase sensitization to toxicants.

Response of Solanum lycopersicum L. to diclofenac – Impacts on the plant’s antioxidant mechanisms

One emerging problem that recently has become a vastly acknowledged topic of concern is the environmental contamination by pharmaceuticals. Diclofenac (DCF) is one of the most common pharmaceuticals found, due to its high utilization and low removal rate in wastewater treatment processes. In this work, Solanum lycopersicum L. was used as a model to unravel how DCF contamination can affect crops, focusing on the internal mechanisms triggered by this exposure. For this purpose, plants were exposed to two different DCF concentrations (0.5 mg L−1 and 5 mg L−1). Results obtained here point towards a loss of shoot performance when plants were exposed to very high concentrations of DCF, but no delay or loss of yield in the flowering and fruit stages were ascribed to DCF contamination. Our data shows that a state of oxidative stress due to high reactive oxygen species accumulation was associated with this contamination, with very high DCF levels leading to a rise of lipid peroxidation, possibly accentuated by the inhibition of ROS-scavenging enzymes and unable to be counteracted by the visible upregulation of proline and the thiol-based redox network. Overall, these results allow to infer that in the current environmental context, no noticeable negative effects should be associated with the presence of DCF in soils where this crop is cultivated. However, the oxidative stress and lower biomass associated with the highest concentration are alarming, since DCF levels in the environment are continuously increasing and further measures are necessary to assess this problematic.

Development and Validation of a Bioanalytical HPLC Method for Quantification of Valsartan in Human Plasma and its Application in Pharmacokinetics Studies

Simple, sensitive, accurate and validated bioanalytical liquid chromatographic procedure coupled with fluorescence detector for estimation of valsartan in presence of diclofenac as internal standard (IS) in plasma has been established. The extraction of valsartan and IS was achieved using methyl tertiary-butyl ether: ethyl acetate (2:8) mixture and the extracted samples were separated on Waters, Xterra- C18 (150 x 4.6 mm, 5 μm) column with 1.0 mL/min flow rate. The detection has been performed at excitation and emission λ at 255 nm and 374 nm, respectively. The elution of the analytes was achieved using acetonitrile: phosphate buffer containing 1 g/L sodium salt of 1-Hexane sulphonic acid with pH adjusted to 2.5 with orthophosphoric acid (52:48, v/v) as an eluting system. The standard curves were linear from 5.0-4000.0 ng/ mL. Method validation has been prepared as per FDA guidelines. Good extraction recoveries of 87.22 % and 99.33 % for valsartan and IS have been acquired, respectively. The achieved values of accuracy and precision were in the adequate range. The proposed procedure was conducted effectively for analysis of valsartan after a single oral administration of one tablet of Tareg tablets ® (80 mg valsartan) to six healthy volunteers.

Fabricating I doped TiO2 photoelectrode for the degradation of diclofenac: Performance and mechanism study

The presence of diclofenac (DCF) in water has developed into a severe environmental problem due to its pernicious effects on animals and human being. Herein, a photoelectrocatalytic (PEC) method using I doped TiO2 (I-TiO2) photoelectrode has been developed, which exhibited excellent visible activity in the degradation of DCF as well as pharmaceutical wastewater. The Langmuir Hinshelwood kinetic model, active species trapping experiments and analysis of intermediate products were employed to analyze the PEC degradation mechanism of DCF. The results revealed that the decomposition of DCF was primarily performed on the photoelectrode (I-TiO2) surface, and the degradation process was accompanied by the dechlorination reaction. The degradation of DCF was caused mainly by holes (66.6%) and hydroxyl radicals (27.6%) when Na2SO4 acted as electrolyte. The degradation rate of DCF could be significantly enhanced under the condition of high Cl− concentration (NaCl acted as electrolyte), which was caused by the participation of chloride radicals (Cl) in the PEC process. The Cl was the major factor contributing to the decomposition of DCF, and the holes and hydroxyl radicals played a less important role than Cl under this condition.

Diclofenac release from polycaprolactone 3D matrices produced by electrospinning: influence of fiber structure and composition of the surrounding medium

The release of tritium-labelled diclofenac from 3D matrices produced by electrospinning from polycaprolactone or PCL blends with human serum albumin in hexafluoroisopropanol was studied. Diclofenac is released significantly faster from matrices made from PCL solution with 10% HSA than from matrices made from pure PCL. Diclofenac was also released faster into blood plasma than into PBS, and the presence of diclofenac in the medium reduced the release rate, as indicated by experiments with regular medium replacement. Thus, diclofenac release depends on the composition of electrospinning solution, the nature of the external medium and the diclofenac concentration in the external medium.

Electrochemical oxidation of acetaminophen in the presence of diclofenac and piroxicam - Synthesis of new derivatives and kinetic investigation of toxic quinone imine/drugs interactions

In this research, electrochemical oxidation of acetaminophen was investigated in the presence of steroidal anti-inflammatory drugs diclofenac and piroxicam by cyclic voltammetry and controlled-potential coulometry techniques. The results indicated that N-acetyl-p-benzoquinone-imine (NAPQI), produced by electrooxidation of acetaminophen, reacts with diclofenac and piroxicam, via the Michael addition reaction. Corresponding products were electrochemically synthesized in aqueous solutions using a carbon electrode in an undivided cell. These products were identified by spectroscopic methods (FT-IR, 1HNMR, 13CNMR and MS). The homogeneous rate constants (kobs), based on the suggested electrode mechanism (i.e. EC), were estimated by comparing the experimental cyclic voltammetric responses with those digital simulated results. A comparison of the estimated kobs for the reaction of electrochemically generated N-acetyl-p-benzoquinone-imine (NAPQI) with steroidal anti-inflammatory drugs revealed that kobs (piroxicam) > kobs(diclofenac) at biological pH.

Diclofenac biodegradation by newly isolated Klebsiella sp. KSC: Microbial intermediates and ecotoxicological assessment

The presence of diclofenac, a frequently used analgesic drug in the environment, can be harmful to a variety of organisms like algae, crustaceans and fish. In this study, a bacterial strain of Klebsiella sp. KSC (Gen Bank, accession number KX500307) able to biodegrade high concentration of diclofenac was isolated from livestock soil and identified. The exposure of Klebsiella sp. KSC to 70 mg/L of diclofenac resulted in diclofenac mineralization after 72 h. This is the first study that points out substantial biodegradation of high concentration of diclofenac (70 mg/L) in less than 72 h; and this shows the potential of this strain to be bioaugmented in contaminated sites or to bioreactors. The chemical structure of twelve biotransformation products of diclofenac are proposed. Its transformation pathway may involve hydroxylation, dehydroxylation, decarboxylation and dechlorination of the central ring of diclofenac. Acute ecotoxicity assay with Vibrio fischeri test showed that the main biotransformation product (TP298) of diclofenac by Klebsiella sp. KSC was less toxic than the parent compound.

Influence of photolabile pharmaceuticals on the photodegradation and toxicity of fluoxetine and fluvoxamine.

Pharmaceuticals in the aquatic environment may be decomposed by abiotic and biotic factors. Photodegradation is the most investigated abiotic process, as it occurs in the natural environment and may be applied in wastewater treatment technology. Although pharmaceuticals are detected in effluents and surface water in a mixture, the photodegradation process is mainly evaluated with single compounds. The photodegradation of fluoxetine (FLU) and fluvoxamine (FLX) in the presence of diclofenac (DCF) and triclosan (TCS) was investigated with HPLC and bioassay. FLU did not degrade under UV-Vis irradiation in SunTest CPS+ either with or without the tested additives, although small amounts of desmethyl fluoxetine and 4-(trifluoromethyl)phenol were formed. In contrast, during irradiation, FLX isomerized to cis-FLX. This process was enhanced by DCF and TCS, but to a lesser degree than by humic acids. Thus, the presence and composition of the matrix should be considered in the environmental risk assessment of pharmaceuticals. As the toxicity of the tested solutions depended only on the concentration of the tested drugs, it was suggested that the biological activity of the photodegradation products was lower than that of the parent compounds.

Potential of the strain Raoultella sp. KDF8 for removal of analgesics.

The bacterial strain KDF8 capable of growth in the presence of diclofenac and codeine analgesics was obtained after chemical mutagenesis of nature isolates from polluted soils. The strain KDF8 was identified as Raoultella sp. based on its morphology, biochemical properties, and 16S rRNA gene sequence. It was deposited in the Czech Collection of Microorganisms under the number CCM 8678. A growing culture efficiently removed diclofenac (92% removal) and partially also codeine (about 30% degradation) from culture supernatants within 72h at 28°C. The degradation of six analgesics by the whole cell catalyst was investigated in detail. The maximum degradation of diclofenac (91%) by the catalyst was achieved at pHINI of 7 (1g/L diclofenac). The specific removal rate at high concentrations of diclofenac and codeine increased up to 16.5mg/gCDW per h and 5.1mg/gCDW per h, respectively. HPLC analysis identified 4'-hydroxydiclofenac as a major metabolite of diclofenac transformation and 14-hydroxycodeinone as codeine transformation product. The analgesics ibuprofen and ketoprofen were also removed, albeit to a lower extent of 3.2 and 2.0mg/gCDW per h, respectively. Naproxen and mefenamic acid were not degraded.

Modulation by NADPH oxidase of the chronic cardiovascular and autonomic interaction between cyclosporine and NSAIDs in female rats

Cyclosporine (CSA) and nonsteroidal antiinflammatory drugs (NSAIDs) are used together to manage arthritic disorders with an immune component. Previous reports showed contrasting effects for NSAIDs on CSA nephrotoxicity and acute elevations in blood pressure. Both effects were ameliorated or exaggerated after selective cyclooxygenase-2 (COX2) and nonselective COX inhibition, respectively. Here we investigated: (i) the interaction of CSA with NSAIDs possessing variable COX1/COX2 selectivities on hemodynamic, left ventricular (LV) and cardiac autonomic and histologic profiles, and (ii) role of NADPH-oxidase (NOX)/Rho-kinase (ROCK) pathway in the interaction. Female rats were pre-instrumented with femoral catheters and treated for 10 days with CSA (25mg/kg/day), diclofenac (nonselective NSAIDs, 1mg/kg/day), celecoxib (COX2 inhibitor, 10mg/kg/day), or their combinations. CSA-mediated hypertension was maintained upon co-administration of either NSAID whereas the concomitant reductions in time- and frequency-domain indices of heart rate variability (HRV) were accentuated in presence of diclofenac but not celecoxib. The isovolumic relaxation time (Τau), a measure of diastolic function, was reduced by all regimens whereas LV contractility (dP/dtmax) remained unaffected. The CSA/diclofenac regimen, but not individual treatments, increased cardiac NOX2 expression and caused more cardiac structural damage. The inhibition of NOX by diphenyleneiodonium reversed CSA/diclofenac-evoked increases in MAP, decreases in HRV and Tau, cardiac structural damage, and increased NOX2 expression. No such effects were observed after ROCK inhibition by fasudil, despite concomitant decreases in NOX2 expression. In conclusion, CSA/diclofenac-treated female rats exhibit exacerbated hemodynamic, autonomic, LV, and histopathologic disturbances via ROCK-independent NOX2 upregulation.

Opposite Modulatory Effects of Selective and Non-Selective Cyclooxygenase Inhibition on Cardiovascular and Autonomic Consequences of Cyclosporine in Female Rats.

Non-steroidal anti-inflammatory drugs (NSAIDs) and the immunosuppressant drug cyclosporine (CSA) are concurrently administered in arthritis. Here, we investigated whether diclofenac (non-selective inhibitor of cyclooxygenase-1 and 2, COX-1/COX-2), celecoxib (selective COX-2 inhibitor) or SC560 (selective COX-1 inhibitor) interact variably with haemodynamic effects of CSA in conscious female rats. Changes caused by CSA (10 mg/kg i.v.) in blood pressure (BP), heart rate (HR), HR variability (HRV) and myocardial contractility were assessed in the absence and presence of individual NSAIDs (10 mg/kg each). Compared with vehicle values, CSA caused immediate and sustained (i) increases in BP and decreases in pulse pressure index (PPI) and HR, (ii) elevations in left ventricular (LV) contractility (dP/dtmax ) and isovolumic relaxation constant (Τau) and (iii) increases in the time- and frequency-domain indices of HRV and shifted the cardiac sympathovagal balance towards parasympathetic dominance. CSA hypertension was abolished in rats pre-treated with celecoxib and intensified in the presence of diclofenac or SC560. Alternatively, the CSA-evoked decreases in PPI, increases in HRV indices and cardiac parasympathetic dominance were blunted by celecoxib in contrast to no effect for diclofenac or SC560. Similarly, celecoxib, but not other NSAIDs, eliminated the elevated LV contractility (dP/dtmax ) and isovolumic relaxation constant (Τau, τ), which reflect cardiac systolic and diastolic function, respectively, that accompanied the CSA-induced pressure load. The data underscore the preferential capacity of selective COX-2 inhibition by celecoxib to mitigate CSA hypertension and consequent alterations in cardiac performance and autonomic balance. By contrast, CSA effects are preserved or even exacerbated after COX-1 inhibition.

Coping with seawater acidification and the emerging contaminant diclofenac at the larval stage: A tale from the clam Ruditapes philippinarum

Seawater acidification could alter the susceptibility of marine organisms to emerging contaminants, such as pharmaceuticals. In this study, the combined effects of seawater acidification and the non-steroidal anti-inflammatory drug diclofenac on survival, growth and oxidative stress-related parameters (catalase activity and lipid peroxidation) in the larvae of the Manila clam Ruditapes philippinarum were investigated for the first time. An experimental flow-through system was set up to carry out a 96-h exposure of clam larvae. Two pH levels (pH 8.0, the control, and pH 7.8, the predicted pH by the end of this century) were tested with and without diclofenac (0.5 μg/L). After 4 days, mortality was dramatically higher under reduced pH, particularly in the presence of diclofenac (62% of the larvae dead). Shell morphology was negatively affected by both acidification and diclofenac from the first day of exposure. The percentage of abnormal larvae was always higher at pH 7.8 than in controls, peaking at 98% in the presence of diclofenac after 96 h. Instead, shell length, shell height or the ratio of these values were only negatively influenced by reduced pH throughout the whole experiment.After 96 h, catalase activity was significantly increased in all larvae kept at pH 7.8, whereas no significant difference in lipid peroxidation was found among the treatments. This study demonstrates a high susceptibility of R. philippinarum larvae to a slight reduction in seawater pH. Furthermore, the results obtained highlight that acidification enhances the sensitivity of clam larvae to environmentally relevant concentrations of diclofenac.

Comparative assessment of diclofenac removal from water by different microalgae strains

Diclofenac has recently been included in the first watch list of substances to be monitored in all member states so as to evaluate its future inclusion in the priority substances list by the Water Framework Directive. Therefore, in view of upcoming limitations on diclofenac discharge, the objective of this work was to assess its removal from water by a microalgae-based treatment. Moreover, considering microalgae application in wastewater treatment, it was aimed to verify if their nutrient removal capacity was affected by the presence of diclofenac. For a comparison purpose, three different microalgae strains, namely Chlorella sorokiniana, Chlorella vulgaris and Scenedesmus obliquus, were cultured in photobioreactors under identical controlled conditions. For the three strains, the addition of diclofenac meant an organic carbon source and provided higher biomass concentration. C. sorokiniana was the strain showing the largest increase of growth rate and microalgae density, which were above 25% and 31%, respectively, compared with the positive control. However, S. obliquus showed the highest efficiency in the removal of diclofenac (>79%) and nutrients (>87% nitrates, >99% phosphates) per litre and per gram of biomass. These results pointed to the potential application of microalgae for the removal of pharmaceuticals in the bioremediation of wastewater.

Removal of diclofenac and sulfamethoxazole from synthetic municipal waste water in microcosm downflow constructed wetlands: Start-up results

ABSTRACTThe objectives of this study were to investigate the start-up removal of pharmaceutical compounds diclofenac and sulfamethoxazole in microcosm downflow constructed wetlands and their effect on the performance of the studied constructed wetlands, and also to assess the effect of plants on the removal of these compounds. The experimental system that was used in this 86-day experiment consisted of 24 columns filled up to 70 cm with predominantly sandy material. Four types of columns were used (six replicates) depending on the presence of plants (Phalaris arundinacea L. var. picta L.) and the presence of pharmaceutical compounds in the influent. The influent was synthetic municipal waste water to which a mixture of 5 mg/L of diclofenac and 5 mg/L of sulfamethoxazole was added. The observed removal of diclofenac was moderate (approx. 50%) and the removal of sulfamethoxazole was relatively low (24–30%). It was found that the removal of diclofenac and sulfamethoxazole was not affected by the vegetation. The presence of diclofenac and sulfamethoxazole in the influent had significant effect on the effluent concentration of N-NO3 and the water loss in the columns, which in both cases were lower than in the control columns. The scope for further research was discussed.

Application of multivariate optimization to electrochemical determination of methyldopa drug in the presence of diclofenac at a nanostructured electrochemical sensor

In the present paper, for the first time multivariate optimization based on central composite rotatable design (CCRD) coupled with response surface methodology (RSM) was used to optimize all effective experimental variables for determination of methyldopa (MD). A simple and sensitive carbon nanocomposite electrode has been developed for the electrochemical trace determination of MD in the presence of diclofenac (DCF) in phosphate buffer solution. This sensor was designed by incorporation of Cu(OH)2 nanoparticles into the carbon paste electrode (CPE). The nanocomposite was characterized by scanning electron microscopy (SEM). Applicability and suitability of Cu(OH)2 nanoparticles/CPE for determination of MD were checked in details using different methods such as differential pulse voltammetry (DPV), electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV) and chronoamperometry (CA). Using DPV, the prepared sensor showed good sensitivity for the determination of MD in the presence of DCF in the range of 2.0–450.1μM, with a detection limit (S/N=3) of 0.61μM. The proposed electrochemical sensor was also used for the determination of MD in some real samples.

Inhibition of islet amyloid polypeptide aggregation and associated cytotoxicity by nonsteroidal anti-inflammatory drugs.

Nonsteroidal anti-inflammatory drugs (NSAIDs) constitute an important pharmacotherapeutic class that, over the past decade, have expanded in application to a panoply of medical conditions. They have been tested for neurodegenerative diseases such as Alzheimer's to reduce inflammation and also in the attempt to abrogate amyloid deposition. However, the use of NSAIDs as aggregation inhibitors has not been extensively studied in pancreatic amyloid deposition. Pancreatic amyloidosis involves the misfolding of islet amyloid polypeptide (IAPP) and contributes to the progression of type-2 diabetes in humans and felines. To ascertain their antiamyloidogenic activity, several NSAIDs were tested using fluorometric thioflavin-T assays, circular dichroism, photo-induced cross-linking assays, and cell culture. Celecoxib, diclofenac, indomethacin, meloxicam, niflumic acid, nimesulide, phenylbutazone, piroxicam, sulindac, and tenoxicam reduced fibrillization at a molar ratio of 1:10. The circular dichroism spectra of diclofenac, piroxicam, and sulindac showed characteristic spectral signatures found in predominantly α-helical structures. The oligomerization of human IAPP was abrogated with diclofenac and sulindac at a molar ratio of 1:5. The cytotoxic effects of pre-incubated human IAPP on cultured INS-1 cells were noticeably reduced in the presence of diclofenac, meloxicam, phenylbutazone, sulindac, and tenoxicam at a molar ratio of 1:10. Our results demonstrate that NSAIDs can provide chemical scaffolds to generate new and promising antiamyloidogenic agents that can be used alone or as a coadjuvant therapy.

Divergent Effects for Celecoxib and Diclofenac on Hemodynamic and Left Ventricular Actions of Cyclosporine in Female Rats

The combined use of cyclosporine (CSA), a potent immunosuppressant, and nonsteroidal antiinflammatory drugs (NSAIDs) in arthritis is a common clinical practice. In this study, we evaluated whether diclofenac (nonselective COX‐1/COX‐2 inhibitor) and celecoxib (selective COX‐2 inhibitor) interact variably with CSA hemodynamics in conscious female rats. Changes evoked by acutely administered CSA in blood pressure (BP), heart rate (HR), HR variability (HRV), and myocardial contractility were assessed in the absence and presence of diclofenac or celecoxib. Compared with vehicle values, CSA (10 mg/kg i.v.) caused immediate and sustained increases in BP and decreases in the pulse pressure index and HR. CSA also increased the time and frequency domain indices of HRV and shifted the cardiac sympathovagal balance towards parasympathetic dominance. The pressor effect of CSA was intensified and abolished in rats pretreated i.v. with diclofenac (10 mg/kg) and celecoxib (10 mg/kg), respectively. The autonomic and PPI effects of CSA were blunted in the presence of celecoxib in contrast to no effect for diclofenac. We also report that celecoxib, but not diclofenac, eliminated the elevated LV contractility (dP/dtmax) and isovolumic relaxation constant (Τau, τ), which reflects the cardiac diastolic function, that accompanied the CSA‐evoked pressure load. The data highlight the preferential capacity of celecoxib to mitigate the pressor effect of CSA and consequent alterations in cardiac performance and autonomic balance in female rats.Supported by the ALEX‐REP Grant Fund, Alexandria University, Egypt (Grant HLTH‐13‐01).

Exercise mitigates diclofenac‐induced liver mitochondrial dysfunction

Several strategies have been developed to counteract liver injury as a consequence of nonsteroid anti-inflammatory drugs toxicity. Here, we aimed to determine whether physical exercise results in liver mitochondrial protection against in vitro diclofenac toxicity. Male adult Sprague-Dawley rats were divided into sedentary, 12-week endurance training (ET) and voluntary activity (VPA). In vitro liver mitochondrial function as assessed by oxygen consumption, transmembrane electric potential (ΔΨ) and susceptibility to the mitochondrial permeability transition pore (MPTP) was evaluated in the absence and presence of diclofenac. Mitochondrial oxidative stress markers [MnSOD, aconitase, -SH and MDA, SIRT3, p66shc(Ser36)/p66shc ratio] and apoptotic signalling (caspases 3, 8 and 9, Bax, Bcl-2 and CypD) were assessed. Content of OXPHOS components and qualitative liver morphological evaluation were assessed. Despite no effects of ET and VPA on basal liver mitochondrial oxygen consumption or ΔΨ endpoints, exercised animals showed lower susceptibility to MPTP. Diclofenac-induced decrease in ΔΨ, increased state 4 respiration and susceptibility to MPTP opening were all prevented by exercise. Under untreated conditions, VPA group showed higher aconitase activity, while ET decreased MDA and increased Bax content. VPA decreased p66shc(Ser36), complex III and V OXPHOS subunits. Both ET and VPA increased complex IV OXPHOS subunit, and SIRT3 and Bcl-2 content and decreased caspase 9 activity. Unexpectedly, ET and VPA decreased ANT. Both chronic physical exercise models augmented the resistance to in vitro diclofenac-induced mitochondrial alterations, including increased MPTP susceptibility, possibly by modulating oxidative stress and MPTP regulators.