The Citrus Flavonoid Hesperetin Has an Inadequate Anti-Arrhythmic Profile in the ΔKPQ NaV1.5 Mutant of the Long QT Type 3 Syndrome.

Type 3 long QT syndromes (LQT3) are associated with arrhythmogenic gain-of-function mutations in the cardiac voltage-gated Na+ channel (hNaV1.5). The citrus flavanone hesperetin (HSP) was previously suggested as a template molecule to develop new anti-arrhythmic drugs, as it blocks slowly-inactivating currents carried by the LQT3-associated hNaV1.5 channel mutant R1623Q. Here we investigated whether HSP also has potentially beneficial effects on another LQT3 hNaV1.5 channel variant, the ΔKPQ, which is associated to lethal ventricular arrhythmias. We used whole-cell patch-clamp to record Na+ currents (INa) in HEK293T cells transiently expressing hNaV1.5 wild type or ΔKPQ mutant channels. HSP blocked peak INa and the late INa carried by ΔKPQ mutant channels with an effective concentration of ≈300 µM. This inhibition was largely voltage-independent and tonic. HSP decreased the rate of inactivation of ΔKPQ channels and, consequently, was relatively weak in reducing the intracellular Na+ load in this mutation. We conclude that, although HSP has potential value for the treatment of the R1623Q LQT3 variant, this compound is inadequate to treat the LQT3 associated to the ΔKPQ genetic variant. Our results underscore the precision medicine rationale of better understanding the basic pathophysiological and pharmacological mechanisms to provide phenotype- genotype-directed individualization of treatment.

Mammalian Transient Receptor Potential TRPA1 Channels: From Structure to Disease.

The transient receptor potential ankyrin (TRPA) channels are Ca2+-permeable nonselective cation channels remarkably conserved through the animal kingdom. Mammals have only one member, TRPA1, which is widely expressed in sensory neurons and in non-neuronal cells (such as epithelial cells and hair cells). TRPA1 owes its name to the presence of 14 ankyrin repeats located in the NH2 terminus of the channel, an unusual structural feature that may be relevant to its interactions with intracellular components. TRPA1 is primarily involved in the detection of an extremely wide variety of exogenous stimuli that may produce cellular damage. This includes a plethora of electrophilic compounds that interact with nucleophilic amino acid residues in the channel and many other chemically unrelated compounds whose only common feature seems to be their ability to partition in the plasma membrane. TRPA1 has been reported to be activated by cold, heat, and mechanical stimuli, and its function is modulated by multiple factors, including Ca2+, trace metals, pH, and reactive oxygen, nitrogen, and carbonyl species. TRPA1 is involved in acute and chronic pain as well as inflammation, plays key roles in the pathophysiology of nearly all organ systems, and is an attractive target for the treatment of related diseases. Here we review the current knowledge about the mammalian TRPA1 channel, linking its unique structure, widely tuned sensory properties, and complex regulation to its roles in multiple pathophysiological conditions.

The citrus flavanone hesperetin preferentially inhibits slow-inactivating currents of a long QT syndrome type 3 syndrome Na+ channel mutation.

BACKGROUND AND PURPOSE: The citrus flavanone hesperetin has been proposed for the treatment of several human pathologies, but its cardiovascular actions remain largely unexplored. Here, we evaluated the effect of hesperetin on cardiac electrical and contractile activities, on aortic contraction, on the wild-type voltage-gated NaV 1.5 channel, and on a channel mutant (R1623Q) associated with lethal ventricular arrhythmias in the long QT syndrome type 3 (LQT3). EXPERIMENTAL APPROACH: We used cardiac surface ECG and contraction force recordings to evaluate the effects of hesperetin in rat isolated hearts and aortic rings. Whole-cell patch clamp was used to record NaV 1.5 currents (INa ) in rat ventricular cardiomyocytes and in HEK293T cells expressing hNaV 1.5 wild-type or mutant channels. KEY RESULTS: Hesperetin increased the QRS interval and heart rate and decreased the corrected QT interval and the cardiac and aortic contraction forces at concentrations equal or higher than 30 µmol·L-1 . Hesperetin blocked rat and human NaV 1.5 channels with an effective inhibitory concentration of ≈100 µmol·L-1 . This inhibition was enhanced at depolarized holding potentials and higher stimulation frequency and was reduced by the disruption of the binding site for local anaesthetics. Hesperetin increased the rate of inactivation and preferentially inhibited INa during the slow inactivation phase, these effects being more pronounced in the R1623Q mutant. CONCLUSIONS AND IMPLICATIONS: Hesperetin preferentially inhibits the slow inactivation phase of INa , more markedly in the mutant R1623Q. Hesperetin could be used as a template to develop drugs against lethal cardiac arrhythmias in LQT3.

Silica nanoparticles inhibit the cation channel TRPV4 in airway epithelial cells.

BACKGROUND: Silica nanoparticles (SiNPs) have numerous beneficial properties and are extensively used in cosmetics and food industries as anti-caking, densifying and hydrophobic agents. However, the increasing exposure levels experienced by the general population and the ability of SiNPs to penetrate cells and tissues have raised concerns about possible toxic effects of this material. Although SiNPs are known to affect the function of the airway epithelium, the molecular targets of these particles remain largely unknown. Given that SiNPs interact with the plasma membrane of epithelial cells we hypothesized that they may affect the function of Transient Receptor Potential Vanilloid 4 (TRPV4), a cation-permeable channel that regulates epithelial barrier function. The main aims of this study were to evaluate the effects of SiNPs on the activation of TRPV4 and to determine whether these alter the positive modulatory action of this channel on the ciliary beat frequency in airway epithelial cells. RESULTS: Using fluorometric measurements of intracellular Ca2+ concentration ([Ca2+]i) we found that SiNPs inhibit activation of TRPV4 by the synthetic agonist GSK1016790A in cultured human airway epithelial cells 16HBE and in primary cultured mouse tracheobronchial epithelial cells. Inhibition of TRPV4 by SiNPs was confirmed in intracellular Ca2+ imaging and whole-cell patch-clamp experiments performed in HEK293T cells over-expressing this channel. In addition to these effects, SiNPs were found to induce a significant increase in basal [Ca2+]i, but in a TRPV4-independent manner. SiNPs enhanced the activation of the capsaicin receptor TRPV1, demonstrating that these particles have a specific inhibitory action on TRPV4 activation. Finally, we found that SiNPs abrogate the increase in ciliary beat frequency induced by TRPV4 activation in mouse airway epithelial cells. CONCLUSIONS: Our results show that SiNPs inhibit TRPV4 activation, and that this effect may impair the positive modulatory action of the stimulation of this channel on the ciliary function in airway epithelial cells. These findings unveil the cation channel TRPV4 as a primary molecular target of SiNPs.

Direct actions of naringin on rat cardiac and vascular smooth muscle / Acciones directas de la naringina sobre los músculos cardíaco y liso vascular de rata

Naringin (NRG) is a flavanone glycoside present in grapefruit juice. Its biological activity has been only partially characterized and little is known about its potential effects in the cardiovascular system. We studied the effects of NRG on the electrical and contractile activities of isolated rat hearts and on the contraction of rat abdominal aortic rings. NRG exerted a negative inotropic action in hearts with an IC50 of 72.5 umol/L but its effects on heart rate and surface electrogram were minimal. Surprisingly, NRG (10-100 umol/L) was able to increase tension in aortic rings contracted by isotonic KCl or phenylephrine. This action of NRG was also evident in aortic rings in basal (resting) conditions but it was absent when resting aortic rings were previously perfused with ryanodine (30 umol/L). Our results indicate that NRG has direct actions on cardiac and vascular smooth muscles that should be taken into account when considering this molecule either as a dietetic supplement or as a template to develop therapeutic agents for human diseases.

La naringina (NRG) es un glicósido de flavanona que se encuentra presente en el jugo de toronja. Su actividad biológica ha sido solo parcialmente caracterizada y poco se conoce acerca de sus efectos sobre el sistema cardiovascular. En la presente investigación estudiamos los efectos de la NRG sobre las actividades eléctrica y contráctil de corazones aislados de rata y sobre la contracción de anillos de aorta abdominal de rata. La NRG ejerció una acción inotropo-negativa en corazones con una IC50 de 72.5 umol/L pero sus efectos sobre la frecuencia cardíaca y el electrograma de superficie fueron mínimos. Sorpresivamente, la NRG (10-100 umol/L) incrementó la tensión en anillos de aorta contraídos por KCl isotónico o fenilefrina. Esta acción de la NRG ocurrió también en anillos de aorta en condiciones basales (en reposo) pero estuvo ausente cuando los anillos de aorta fueron previamente perfundidos con ryanodina (30 umol/L). Nuestros resultados indican que la NRG tiene acciones directas sobre los músculos cardíaco y liso vascular que deben tenerse en cuenta al considerar esta molécula como suplemento dietético o como plantilla para el desarrollo de agentes terapéuticos para el tratamiento de enfermedades en humanos.

Cinnamaldehyde inhibits L-type calcium channels in mouse ventricular cardiomyocytes and vascular smooth muscle cells.

Cinnamaldehyde (CA), a major component of cinnamon, is known to have important actions in the cardiovascular system, including vasorelaxation and decrease in blood pressure. Although CA-induced activation of the chemosensory cation channel TRPA1 seems to be involved in these phenomena, it has been shown that genetic ablation of Trpa1 is insufficient to abolish CA effects. Here, we confirm that CA relaxes rat aortic rings and report that it has negative inotropic and chronotropic effects on isolated mouse hearts. Considering the major role of L-type Ca(2+) channels in the control of the vascular tone and cardiac contraction, we used whole-cell patch-clamp to test whether CA affects L-type Ca(2+) currents in mouse ventricular cardiomyocytes (VCM, with Ca(2+) as charge carrier) and in mesenteric artery smooth muscle cells (VSMC, with Ba(2+) as charge carrier). We found that CA inhibited L-type currents in both cell types in a concentration-dependent manner, with little voltage-dependent effects. However, CA was more potent in VCM than in VSMC and caused opposite effects on the rate of inactivation. We found these divergences to be at least in part due to the use of different charge carriers. We conclude that CA inhibits L-type Ca(2+) channels and that this effect may contribute to its vasorelaxing action. Importantly, our results demonstrate that TRPA1 is not a specific target of CA and indicate that the inhibition of voltage-gated Ca(2+) channels should be taken into account when using CA to probe the pathophysiological roles of TRPA1.

Acciones cardiovasculares del ibuprofeno / Ibuprofen cardiovascular actions

El ibuprofeno (IB) es un antiinflamatorio no esteroideo (AINE) de amplio uso por su alta efectividad y buen margen de seguridad. Sin embargo, poco se conoce de sus posibles acciones cardiovasculares. Algunas evidencias clínicas sugieren que este AINE pudiera tener efectos adversos sobre el sistema cardiovascular. El objetivo de esta investigación fue estudiar las posibles acciones colaterales del ibuprofeno sobre corazón y músculo liso vascular, tomando como patrón de comparación el ácido flufenámico (AF), fármaco con probada acción inotropo negativa. Se utilizó la técnica clásica de corazón de rata aislado y perfundido (Langendorff), registrando electrograma superficial y la fuerza de contracción. También se estudió el efecto sobre la contracción de aorta abdominal de rata inducida por KCl isotónico y por fenilefrina (10 µmol/L). En comparación con el AF (IC50=9,5 µmol/L), el IB tuvo un pobre efecto inotrópico negativo (IC20=30 µmol/L). A la concentración máxima utilizada (100 µmol/L), el IB fue menos efectivo que el AF en reducir el intervalo QT (25 ± 7 ms vs. 60 ± 15 ms; N ³ 5) y alargar el intervalo RR (60 ± 10 ms vs. 145 ± 20 ms; N ³ 5). Mientras que el AF no tuvo acción sobre la contracción aórtica inducida por KCl o por fenilefrina, el IB provocó una vasorrelajación de » 30 por ciento de la contracción aórtica inducida por KCl o por fenilefrina aunque solo a la concentración máxima (100 µmol/L). Estos resultados sugieren que las acciones cardiovasculares directas del IB son mínimas lo cual contribuye al buen margen de seguridad para su uso en clínica en pacientes sin enfermedad cardiovascular

Ibuprofen (IB) is a non steroidal anti-inflammatory drug (NSAID) widely used because of its high effectivity and good safety margin. However, little is known about its possible cardiovascular actions. Some evidences in clinics suggest that this NSAID could have adverse side effects on the cardiovascular system. The aim of this investigation was to study the possible side effects of IB on the heart and vascular smooth muscle, taking as a reference the flufenamic acid (FA) a drug with known negative inotropic action. We used the isolated perfused rat heart (Langendorff) and recorded the surface electrogram and the force of contraction. We also studied the effects of IB on the KCl- or fenilephrine- (10 µmol/L) induced contraction of rat abdominal aorta. Compared to FA (IC50 = 9.5 µmol/L), IB showed a small negative inotropic effect (IC20 = 30 µmol/L). At the maximal concentration used (100 µmol/L), IB was less effective than FA in reducing the QT interval (45 ± 10 ms vs. 60 ± 15 ms; N ³ 5) and prolonging the RR interval (60 ± 10 ms vs. 145 ± 20 ms; N ³ 5). While FA had no effect on the aortic contraction (KCl or fenilepinephrine), IB relaxed aortic contraction (KCl or fenilephrine) by » 30 percent but only at the highest concentration (100 µmol/L). The present results suggest that cardiovascular actions of IB are minimal contributing to its good safety margin when used in clinics in patients not suffering from cardiovascular diseases

Acciones cardiovasculares del ibuprofeno / Ibuprofen cardiovascular actions

El ibuprofeno (IB) es un antiinflamatorio no esteroideo (AINE) de amplio uso por su alta efectividad y buen margen de seguridad. Sin embargo, poco se conoce de sus posibles acciones cardiovasculares. Algunas evidencias clínicas sugieren que este AINE pudiera tener efectos adversos sobre el sistema cardiovascular. El objetivo de esta investigación fue estudiar las posibles acciones colaterales del ibuprofeno sobre corazón y músculo liso vascular, tomando como patrón de comparación el ácido flufenámico (AF), fármaco con probada acción inotropo negativa. Se utilizó la técnica clásica de corazón de rata aislado y perfundido (Langendorff), registrando electrograma superficial y la fuerza de contracción. También se estudió el efecto sobre la contracción de aorta abdominal de rata inducida por KCl isotónico y por fenilefrina (10 Ámol/L). En comparación con el AF (IC50=9,5 Ámol/L), el IB tuvo un pobre efecto inotrópico negativo (IC20=30 Ámol/L). A la concentración máxima utilizada (100 Ámol/L), el IB fue menos efectivo que el AF en reducir el intervalo QT (25 ± 7 ms vs. 60 ± 15 ms; N ³ 5) y alargar el intervalo RR (60 ± 10 ms vs. 145 ± 20 ms; N ³ 5). Mientras que el AF no tuvo acción sobre la contracción aórtica inducida por KCl o por fenilefrina, el IB provocó una vasorrelajación de ¼ 30 por ciento de la contracci¾n aórtica inducida por KCl o por fenilefrina aunque solo a la concentración máxima (100 Ámol/L). Estos resultados sugieren que las acciones cardiovasculares directas del IB son mínimas lo cual contribuye al buen margen de seguridad para su uso en clínica en pacientes sin enfermedad cardiovascular(AU)

Ibuprofen (IB) is a non steroidal anti-inflammatory drug (NSAID) widely used because of its high effectivity and good safety margin. However, little is known about its possible cardiovascular actions. Some evidences in clinics suggest that this NSAID could have adverse side effects on the cardiovascular system. The aim of this investigation was to study the possible side effects of IB on the heart and vascular smooth muscle, taking as a "reference" the flufenamic acid (FA) a drug with known negative inotropic action. We used the isolated perfused rat heart (Langendorff) and recorded the surface electrogram and the force of contraction. We also studied the effects of IB on the KCl- or fenilephrine- (10 Ámol/L) induced contraction of rat abdominal aorta. Compared to FA (IC50 = 9.5 Ámol/L), IB showed a small negative inotropic effect (IC20 = 30 Ámol/L). At the maximal concentration used (100 Ámol/L), IB was less effective than FA in reducing the QT interval (45 ± 10 ms vs. 60 ± 15 ms; N ³ 5) and prolonging the RR interval (60 ± 10 ms vs. 145 ± 20 ms; N ³ 5). While FA had no effect on the aortic contraction (KCl or fenilepinephrine), IB relaxed aortic contraction (KCl or fenilephrine) by ¼ 30 percent but only at the highest concentration (100 Ámol/L). The present results suggest that cardiovascular actions of IB are minimal contributing to its good safety margin when used in clinics in patients not suffering from cardiovascular diseases(AU)

Acciones cardiovasculares del ibuprofeno / Ibuprofen cardiovascular actions

El ibuprofeno (IB) es un antiinflamatorio no esteroideo (AINE) de amplio uso por su alta efectividad y buen margen de seguridad. Sin embargo, poco se conoce de sus posibles acciones cardiovasculares. Algunas evidencias clínicas sugieren que este AINE pudiera tener efectos adversos sobre el sistema cardiovascular. El objetivo de esta investigación fue estudiar las posibles acciones colaterales del ibuprofeno sobre corazón y músculo liso vascular, tomando como patrón de comparación el ácido flufenámico (AF), fármaco con probada acción inotropo negativa. Se utilizó la técnica clásica de corazón de rata aislado y perfundido (Langendorff), registrando electrograma superficial y la fuerza de contracción. También se estudió el efecto sobre la contracción de aorta abdominal de rata inducida por KCl isotónico y por fenilefrina (10 µmol/L). En comparación con el AF (IC50=9,5 µmol/L), el IB tuvo un pobre efecto inotrópico negativo (IC20=30 µmol/L). A la concentración máxima utilizada (100 µmol/L), el IB fue menos efectivo que el AF en reducir el intervalo QT (25 ± 7 ms vs. 60 ± 15 ms; N ³ 5) y alargar el intervalo RR (60 ± 10 ms vs. 145 ± 20 ms; N ³ 5). Mientras que el AF no tuvo acción sobre la contracción aórtica inducida por KCl o por fenilefrina, el IB provocó una vasorrelajación de ¼ 30 por ciento de la contracción aórtica inducida por KCl o por fenilefrina aunque solo a la concentración máxima (100 µmol/L). Estos resultados sugieren que las acciones cardiovasculares directas del IB son mínimas lo cual contribuye al buen margen de seguridad para su uso en clínica en pacientes sin enfermedad cardiovascular(AU)

Ibuprofen (IB) is a non steroidal anti-inflammatory drug (NSAID) widely used because of its high effectivity and good safety margin. However, little is known about its possible cardiovascular actions. Some evidences in clinics suggest that this NSAID could have adverse side effects on the cardiovascular system. The aim of this investigation was to study the possible side effects of IB on the heart and vascular smooth muscle, taking as a reference the flufenamic acid (FA) a drug with known negative inotropic action. We used the isolated perfused rat heart (Langendorff) and recorded the surface electrogram and the force of contraction. We also studied the effects of IB on the KCl- or fenilephrine- (10 µmol/L) induced contraction of rat abdominal aorta. Compared to FA (IC50 = 9.5 µmol/L), IB showed a small negative inotropic effect (IC20 = 30 µmol/L). At the maximal concentration used (100 µmol/L), IB was less effective than FA in reducing the QT interval (45 ± 10 ms vs. 60 ± 15 ms; N ³ 5) and prolonging the RR interval (60 ± 10 ms vs. 145 ± 20 ms; N ³ 5). While FA had no effect on the aortic contraction (KCl or fenilepinephrine), IB relaxed aortic contraction (KCl or fenilephrine) by ¼ 30 percent but only at the highest concentration (100 µmol/L). The present results suggest that cardiovascular actions of IB are minimal contributing to its good safety margin when used in clinics in patients not suffering from cardiovascular diseases(AU)