RESUMEN
This paper suggests that ATP release induced by the SARS-CoV-2 virus plays a key role in the genesis of the major symptoms and complications of COVID-19. Infection of specific cells which contain the Angiotensin-Converting Enzyme 2 (ACE2) receptor results in a loss of protection of the Mineralocorticoid Receptor (MR). Local activation by cortisol stimulates the release of ATP initially into the basolateral compartment and then by lysosomal exocytosis from the cell surface. This then acts on adjacent cells. In the nose ATP acts as a nociceptive stimulus which results in anosmia. It is suggested that a similar paracrine mechanism is responsible for the loss of taste. In the lung ATP release from type 2 alveolar cells produces the non-productive cough by acting on purinergic receptors on adjacent neuroepithelial cells and activating, via the vagus, the cough reflex. Infection of endothelial cells results in the exocytosis of WeibelPalade bodies. These contain the Von Willebrand Factor responsible for micro-clotting and angiopoietin-2 which increases vascular permeability and plays a key role in the Acute Respiratory Distress Syndrome. To test this hypothesis this paper reports proof of concept studies in which MR blockade using spironolactone and low dose dexamethasone (SpiDex) was given to PCR-confirmed COVID-19 patients. In 80 patients with moderate to severe respiratory failure 40 were given SpiDex and 40 conventional treatment with high dose dexamethasone (HiDex). There was 1 death in the HiDex group and none in the SpiDex. As judged by clinical, biochemical and radiological parameters there were clear statistically significant benefits of SpiDex in comparison to HiDex. A further 20 outpatients with COVID-19 were given SpiDex. There was no control group and the aim was to demonstrate safety. No adverse effects were noted and no patient became hyperkalaemic. 90% were asymptomatic at 10 days. The very positive results suggest that blockade of the MR can produce major benefit in COVID19 patients. Further larger controlled studies of inpatients and outpatients are required not only for SARS-CoV-2 infection per se but also to determine if this treatment affects the incidence of Long COVID.
Asunto(s)
Anosmia/complicaciones , COVID-19/diagnóstico , COVID-19/terapia , Nocicepción , SARS-CoV-2 , Evaluación de Síntomas , Adenosina Trifosfato/metabolismo , Adulto , Anciano , Angiopoyetina 2/biosíntesis , Enzima Convertidora de Angiotensina 2/biosíntesis , Animales , COVID-19/sangre , Dexametasona/administración & dosificación , Dexametasona/sangre , Dexametasona/uso terapéutico , Células Endoteliales/metabolismo , Femenino , Humanos , Hidrocortisona/metabolismo , Riñón/efectos de los fármacos , Masculino , Persona de Mediana Edad , Modelos Biológicos , Reacción en Cadena de la Polimerasa , Ratas , Receptores de Mineralocorticoides/biosíntesis , Espironolactona/sangre , Factor de von Willebrand/biosíntesisRESUMEN
BACKGROUND: Post-traumatic stress disorder (PTSD) is a trauma-related disorder that is associated with pro-inflammatory activation and neurobiological impairments in the brain and leads to a series of affective-like behaviors. Electroacupuncture (EA) has been proposed as a clinically useful therapy for several brain diseases. However, the potential role of EA treatment in PTSD and its molecular and cellular mechanisms has rarely been investigated. METHODS: We used an established preclinical social defeat stress mouse model to study whether EA treatment modulates PTSD-like symptoms and understand its underlying mechanisms. To this end, male C57BL/6 mice were subjected to repeated social defeat stress (RSDS) for 6 consecutive days to induce symptoms of PTSD and treated with EA at Baihui (GV 20) and Dazhui (GV 14) acupoints. RESULTS: The stimulation of EA, but not needle insertion at Baihui (GV 20) and Dazhui (GV 14) acupoints effectively improved PTSD-like behaviors such as, social avoidance and anxiety-like behaviors. However, EA stimulation at the bilateral Tianzong (SI11) acupoints did not affect the PTSD-like behaviors obtained by RSDS. EA stimulation also markedly inhibited astrocyte activation in both the dorsal and ventral hippocampi of RSDS-treated mice. Using next-generation sequencing analysis, our results showed that EA stimulation attenuated RSDS-enhanced lipocalin 2 expression in the hippocampus. Importantly, using double-staining immunofluorescence, we observed that the increased lipocalin 2 expression in astrocytes by RSDS was also reduced by EA stimulation. In addition, intracerebroventricular injection of mouse recombinant lipocalin 2 protein in the lateral ventricles provoked social avoidance, anxiety-like behaviors, and the activation of astrocytes in the hippocampus. Interestingly, the overexpression of lipocalin 2 in the brain also altered the expression of stress-related genes, including monoamine oxidase A, monoamine oxidase B, mineralocorticoid receptor, and glucocorticoid receptor in the hippocampus. CONCLUSIONS: This study suggests that the treatment of EA at Baihui (GV 20) and Dazhui (GV 14) acupoints improves RSDS-induced social avoidance, anxiety-like behaviors, astrocyte activation, and lipocalin 2 expression. Furthermore, our findings also indicate that lipocalin 2 expression in the brain may be an important biomarker for the development of PTSD-related symptoms.
Asunto(s)
Terapia por Acupuntura , Ansiedad/prevención & control , Electroacupuntura , Hipocampo/metabolismo , Lipocalina 2/fisiología , Derrota Social , Interacción Social , Trastornos por Estrés Postraumático/terapia , Actinas/biosíntesis , Actinas/genética , Puntos de Acupuntura , Animales , Ansiedad/etiología , Prueba de Laberinto Elevado , Conducta Exploratoria , Inyecciones Intraventriculares , Lipocalina 2/biosíntesis , Lipocalina 2/genética , Lipocalina 2/farmacología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos ICR , Monoaminooxidasa/biosíntesis , Monoaminooxidasa/genética , Receptores de Glucocorticoides/biosíntesis , Receptores de Glucocorticoides/genética , Receptores de Mineralocorticoides/biosíntesis , Receptores de Mineralocorticoides/genética , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/farmacología , Trastornos por Estrés Postraumático/etiología , Trastornos por Estrés Postraumático/psicologíaRESUMEN
In this study we tested the hypothesis i) that age-dependent shifts in the excitation-inhibition balance of prefrontal neurons are accelerated by early life stress, a risk factor for the etiology of many psychiatric disorders; and if so, ii) that this process is exacerbated by genetic forebrain-specific downregulation of the mineralocorticoid receptor, a receptor that was earlier found to be a protective factor for negative effects of early life stress in both rodents and humans. In agreement with the literature, an age-dependent downregulation of the excitation-inhibition balance was found both with regard to spontaneous and evoked synaptic currents. The age-dependent shift in spontaneous excitatory relative to inhibitory currents was significantly accelerated by early life stress, but this was not exacerbated by reduction in mineralocorticoid receptor expression. The age-dependent changes in the excitation-inhibition balance were mirrored by similar changes in receptor subunit expression and morphological alterations, particularly in spine density, which could thus potentially contribute to the functional changes. However, none of these parameters displayed acceleration by early life stress, nor depended on mineralocorticoid receptor expression. We conclude that, in agreement with the hypothesis, early life stress accelerates the developmental shift of the excitation-inhibition balance but, contrary to expectation, there is no evidence for a putative protective role of the mineralocorticoid receptor in this system. In view of the modest effect of early life stress on the excitation-inhibition balance, alternative mechanisms potentially underlying the development of psychiatric disorders should be further explored.
Asunto(s)
Potenciales Postsinápticos Excitadores/fisiología , Potenciales Postsinápticos Inhibidores/fisiología , Corteza Prefrontal/metabolismo , Prosencéfalo/metabolismo , Receptores de Mineralocorticoides/biosíntesis , Estrés Psicológico/metabolismo , Factores de Edad , Animales , Animales Recién Nacidos , Femenino , Expresión Génica , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Neuronas/metabolismo , Corteza Prefrontal/patología , Prosencéfalo/patología , Receptores de Mineralocorticoides/genética , Estrés Psicológico/patología , Estrés Psicológico/prevención & controlRESUMEN
The LPA1 receptor, one of the six characterized G protein-coupled receptors (LPA1-6) through which lysophosphatidic acid acts, is likely involved in promoting normal emotional behaviours. Current data suggest that the LPA-LPA1-receptor pathway may be involved in mediating the negative consequences of stress on hippocampal function. However, to date, there is no available information regarding the mechanisms whereby the LPA1 receptor mediates this adaptation. To gain further insight into how the LPA-LPA1 pathway may prevent the negative consequences of chronic stress, we assessed the effects of the continuous delivery of LPA on depressive-like behaviours induced by a chronic restraint stress protocol. Because a proper excitatory/inhibitory balance seems to be key for controlling the stress response system, the gene expression of molecular markers of excitatory and inhibitory neurotransmission was also determined. In addition, the hippocampal expression of mineralocorticoid receptor genes and glucocorticoid receptor genes and proteins as well as plasma corticosterone levels were determined. Contrary to our expectations, the continuous delivery of LPA in chronically stressed animals potentiated rather than inhibited some (e.g., anhedonia, reduced latency to the first immobility period), though not all, behavioural effects of stress. Furthermore, this treatment led to an alteration in the genes coding for proteins involved in the excitatory/inhibitory balance in the ventral hippocampus and to changes in corticosterone levels. In conclusion, the results of this study reinforce the assumption that LPA is involved in emotional regulation, mainly through the LPA1 receptor, and regulates the effects of stress on hippocampal gene expression and hippocampus-dependent behaviour.
Asunto(s)
Conducta Animal , Hipocampo/fisiopatología , Receptores del Ácido Lisofosfatídico/genética , Estrés Psicológico/genética , Estrés Psicológico/psicología , Anhedonia , Animales , Enfermedad Crónica , Corticosterona/sangre , Depresión/psicología , Expresión Génica , Masculino , Ratones , Ratones Endogámicos C57BL , Inhibición Neural , Receptores de Mineralocorticoides/biosíntesis , Receptores de Mineralocorticoides/genética , Estrés Psicológico/fisiopatología , Natación/psicología , Transmisión SinápticaRESUMEN
The administration of glucocorticoids (GCs) for the treatment of traumatic brain injury (TBI) is controversial. Both protective and deleterious effects of GCs on the brain have been reported in previous studies, while the mechanisms are unclear. Most experimental studies have reported glucocorticoid receptor (GR)-mediated deleterious effects after TBI. Sufficient mineralocorticoid receptor (MR) activation was reported to be indispensable for normal function and survival of hippocampal neurons, but changes in MR expression and activation and the roles of MRs in the survival of neurons after TBI remain unclear. We hypothesized that inadequate MR expression and activation caused by TBI aggravates posttraumatic hippocampal apoptosis but that restoration by restoring MRs promotes the survival of neurons. Using a rat controlled cortical impact model, we examined plasma corticosterone, MR expression and activation, neuronal apoptosis in the hippocampus, and spatial memory on day 3 after injury with and without fludrocortisone (1â¯mg/kg) treatment. Plasma corticosterone levels were significantly reduced after TBI. In addition, both MR expression and activation were inhibited. Fludrocortisone treatment significantly increased both the expression and activation of MRs, reduced the number of apoptotic neurons and cell loss in the ipsilateral hippocampus, and subsequently improved spatial memory. Its protective effects were counteracted by the MR antagonist spironolactone. The results suggest that adequate expression and activation of MRs is crucial for the survival of neurons after TBI and that fludrocortisone protects hippocampal neurons via promoting MR expression and activation.
Asunto(s)
Lesiones Traumáticas del Encéfalo/metabolismo , Hipocampo/metabolismo , Receptores de Mineralocorticoides/biosíntesis , Memoria Espacial/fisiología , Animales , Lesiones Traumáticas del Encéfalo/genética , Lesiones Traumáticas del Encéfalo/patología , Expresión Génica , Hipocampo/patología , Masculino , Neuronas/metabolismo , Neuronas/patología , Ratas , Ratas Sprague-Dawley , Receptores de Mineralocorticoides/genéticaRESUMEN
PURPOSE OF REVIEW: To review the latest reports of the contributions of the endothelial mineralocorticoid receptor to endothelial dysfunction and hypertension to begin to determine the clinical potential for this pathway for hypertension treatment. RECENT FINDINGS: Endothelial mineralocorticoid receptor expression is sex-specifically increased in female mice and humans compared with males. Moreover, the expression of endothelial mineralocorticoid receptors is increased by endothelial progesterone receptor activation and naturally occurring fluctuations in progesterone levels (estrous, pregnancy) predict endothelial mineralocorticoid receptor expression levels in female mice. These data follow many previous reports that have indicated that endothelial mineralocorticoid receptor deletion is protective in the development of obesity- and diabetes-associated endothelial dysfunction in female mouse models. These studies have more recently been followed up by reports indicating that both intact endothelial mineralocorticoid receptor and progesterone receptor expression are required for obesity-associated, leptin-mediated endothelial dysfunction in female mice. In addition, the intra-endothelial signaling pathway for endothelial mineralocorticoid receptors to induce dysfunction requires the intact expression of α-epithelial sodium channels (αENaC) in endothelial cells in females. Endothelial mineralocorticoid receptors are sex-specifically upregulated in the vasculature of females, a sex difference which is driven by endothelial progesterone receptor activation, and increased activity of these endothelial mineralocorticoid receptors is a crucial mediator of endothelial dysfunction, and potentially hypertension, in obese female experimental models.
Asunto(s)
Endotelio Vascular/metabolismo , Hipertensión/metabolismo , Obesidad/metabolismo , Receptores de Mineralocorticoides/biosíntesis , Enfermedades Vasculares/metabolismo , Aldosterona/metabolismo , Animales , Modelos Animales de Enfermedad , Células Endoteliales/metabolismo , Endotelio Vascular/fisiopatología , Canales Epiteliales de Sodio/metabolismo , Femenino , Humanos , Leptina/metabolismo , Masculino , Ratones , Antagonistas de Receptores de Mineralocorticoides , Receptores de Progesterona/metabolismo , Factores SexualesRESUMEN
The mineralocorticoid receptor (MR) is a nuclear receptor and part of a large and diverse family of transcription factors that also includes receptors for glucocorticoids, progesterone, androgens, and estrogens. The corticosteroid aldosterone is the physiological activator of the MR in humans and other terrestrial vertebrates; however, its activator is not known in cartilaginous fish, the oldest group of extant jawed vertebrates. Here, we analyzed the ability of corticosteroids and progesterone to activate the full-length MR from the elephant shark (Callorhinchus milii). On the basis of their measured activities, aldosterone, cortisol, 11-deoxycorticosterone, corticosterone, 11-deoxcortisol, progesterone, and 19-norprogesterone are potential physiological mineralocorticoids. However, aldosterone, the physiological mineralocorticoid in humans and other terrestrial vertebrates, is not found in cartilaginous or ray-finned fish. Although progesterone activates MRs in ray-finned fish, progesterone does not activate MRs in humans, amphibians, or alligator, suggesting that during the transition to terrestrial vertebrates, progesterone lost the ability to activate the MR. Both elephant shark MR and human MR are expressed in the brain, heart, ovary, testis, and other nonepithelial tissues, suggesting that MR expression in diverse tissues evolved in the common ancestor of jawed vertebrates. Our data suggest that 19-norprogesterone- and progesterone-activated MR may have unappreciated functions in reproductive physiology.
Asunto(s)
Corticoesteroides/farmacología , Proteínas de Peces/biosíntesis , Progesterona/farmacología , Receptores de Mineralocorticoides/biosíntesis , Tiburones/metabolismo , Espironolactona/farmacología , Activación Transcripcional/efectos de los fármacos , Animales , Proteínas de Peces/genética , Humanos , Especificidad de Órganos/efectos de los fármacos , Receptores de Mineralocorticoides/genética , Tiburones/genéticaRESUMEN
Phthalates are industrial plasticizers and stabilizers commonly found in polyvinyl chloride plastic and consumer products, including food packaging, cosmetics, medical devices, and children's toys. Di-(2-ethylhexyl) phthalate (DEHP), one of the most commonly used phthalates, exhibits endocrine-disrupting characteristics and direct exposure leads to reproductive deficits and abnormalities in anxiety-related behaviors. Importantly, increasing evidence indicates that the impacts of DEHP exposure on reproduction and social behavior persist across multiple generations. In this study, we tested the hypothesis that transgenerational DEHP exposure alters anxiety-like behavior and neural gene expression in both male and female mice. Pregnant CD-1 mice were orally dosed daily with either tocopherol-stripped corn oil or DEHP (20 or 200⯵g/kg/day; 500 or 750â¯mg/kg/day) from gestational day 10.5 until birth to produce the F1 generation. Females from each generation were bred with untreated, unrelated CD-1 males to produce subsequent generations. Behavior and gene expression assays were performed with adult, intact F3 males and females. Transgenerational DEHP exposure increased time spent in the open arm in the elevated plus maze for adult females (750â¯mg/kg/day lineage), but not males. In adult females, we observed a down-regulation of mRNA expression of estrogen receptor 1 in the 200⯵g/kg/day and 500â¯mg/kg/day treatment lineages, mineralocorticoid receptor in the 200⯵g/kg/day lineage, and dopamine receptor 2 in the 20⯵g/kg/day and 750â¯mg/kg/day lineages. In adult males, we found an up-regulation of estrogen receptor 2 in the 20 and 200⯵g/kg/day lineages, and dopamine receptor 1 in the 20⯵g/kg/day and 750â¯mg/kg/day lineages. No hippocampal gene expression modifications were observed in response to treatment. These results implicate dose-specific transgenerational effects on behavior and neural gene expression in adult male and female mice.
Asunto(s)
Amígdala del Cerebelo/efectos de los fármacos , Amígdala del Cerebelo/metabolismo , Ansiedad/genética , Ansiedad/psicología , Dietilhexil Ftalato/toxicidad , Disruptores Endocrinos/toxicidad , Regulación de la Expresión Génica/efectos de los fármacos , Animales , Conducta Animal , Relación Dosis-Respuesta a Droga , Regulación hacia Abajo/efectos de los fármacos , Receptor alfa de Estrógeno/biosíntesis , Receptor alfa de Estrógeno/genética , Femenino , Hipocampo/efectos de los fármacos , Masculino , Exposición Materna , Ratones , Embarazo , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Receptores de Dopamina D2/biosíntesis , Receptores de Dopamina D2/genética , Receptores de Mineralocorticoides/biosíntesis , Receptores de Mineralocorticoides/genética , Caracteres Sexuales , Conducta SocialRESUMEN
Emerging evidence indicates that aldosterone and mineralocorticoid receptors (MRs) are associated with the pathogenesis of erectile dysfunction. However, the molecular mechanisms remain largely unknown. In this study, freshly isolated penile corpus cavernosum tissue from rats was treated with aldosterone, with or without MRs inhibitors. Nuclear factor (NF)-kappa B (NF-κB) activity was evaluated by real-time quantitative PCR, luciferase assay, and immunoblot. The results demonstrated that mRNA levels of the NF-κB target genes, including inhibitor of NF-κB alpha (IκB-α), NF-κB1, tumor necrosis factor-alpha (TNF-α), and interleukin 6 (IL-6), were higher after aldosterone treatment. Accordingly, phosphorylation of p65/RelA, IκB-α, and inhibitor of NF-κB kinase-ß was markedly increased by aldosterone. Furthermore, knockdown of MRs prevented activation of the NF-κB canonical pathway by aldosterone. Consistent with this finding, ectopic overexpression of MRs enhanced the transcriptional activation of NF-κB by aldosterone. More importantly, the MRs antagonist, spironolactone blocked aldosterone-mediated activation of the canonical NF-κB pathway. In conclusion, aldosterone has an inflammatory effect in the corpus cavernosum penis, inducing NF-κB activation via an MRs-dependent pathway, which may be prevented by selective MRs antagonists. These data reveal the possible role of aldosterone in erectile dysfunction as well as its potential as a novel pharmacologic target for treatment.
Asunto(s)
Aldosterona/farmacología , Citocinas/biosíntesis , FN-kappa B/agonistas , Pene/metabolismo , Transducción de Señal/efectos de los fármacos , Animales , Técnicas de Silenciamiento del Gen , Quinasa I-kappa B/antagonistas & inhibidores , Interleucina-6/biosíntesis , Interleucina-6/genética , Masculino , Antagonistas de Receptores de Mineralocorticoides/farmacología , FN-kappa B/genética , Pene/efectos de los fármacos , Proteínas Serina-Treonina Quinasas/antagonistas & inhibidores , ARN Mensajero/biosíntesis , Ratas , Ratas Endogámicas WKY , Receptores de Mineralocorticoides/biosíntesis , Receptores de Mineralocorticoides/genética , Espironolactona/farmacología , Activación Transcripcional , Factor de Necrosis Tumoral alfa/biosíntesis , Quinasa de Factor Nuclear kappa BRESUMEN
Activation of mineralocorticoid receptor (MR) is shown in resistant hypertension including diabetes mellitus. Although protein kinase C (PKC) signaling is involved in the pathogenesis of diabetic complications, an association between PKC and MR is not known. Activation of PKCα and PKCß by TPA (12-O-Tetradecanoylphorbol 13-acetate) increased MR proteins and its transcriptional activities in HEK293-MR cells. In contrast, a high glucose condition resulted in PKCß but not PKCα activation, which is associated with elevation of MR protein levels and MR transcriptional activities. Reduction of endogenous PKCß by siRNA decreased those levels. Interestingly, high glucose did not affect MR mRNA levels, but rather decreased ubiquitination of MR proteins. In db/db mice kidneys, levels of phosphorylated PKCß2, MR and Sgk-1 proteins were elevated, and the administration of PKC inhibitor reversed these changes compared to db/+ mice. These data suggest that high glucose stimulates PKCß signaling, which leads to MR stabilization and its transcriptional activities.
Asunto(s)
Diabetes Mellitus Experimental , Regulación de la Expresión Génica , Glucosa/administración & dosificación , Hipertensión/genética , Proteína Quinasa C beta/genética , ARN/genética , Receptores de Mineralocorticoides/efectos de los fármacos , Animales , Western Blotting , Células Cultivadas , Relación Dosis-Respuesta a Droga , Humanos , Hipertensión/tratamiento farmacológico , Hipertensión/metabolismo , Ratones , Ratones Transgénicos , Proteína Quinasa C beta/biosíntesis , Conejos , Reacción en Cadena en Tiempo Real de la Polimerasa , Receptores de Mineralocorticoides/biosíntesis , Receptores de Mineralocorticoides/genética , Transducción de Señal/efectos de los fármacosRESUMEN
Postsynaptic 5-HT1A receptors (5-HT1AR) play an important role in anxiety and stress, although their contribution is still controversial. Previous studies report that mice overexpressing postsynaptic 5-HT1ARs show no changes in basal anxiety, though the influence of stress conditions has not been addressed yet. In this study, we used this animal model to evaluate the role of 5-HT1ARs in anxiety response after pre-exposure to an acute stressor. Under basal conditions, 5-HT1AR overexpressing animals presented high corticosterone levels and a lower mineralocorticoid/glucocorticoid receptor ratio. After pre-exposure to a single stressor, they showed a high anxiety-like response, associated with a blunted increase in corticosterone levels and higher c-Fos activation in the prefrontal cortex. Moreover, these mice also presented a lack of downregulation of hippocampal long-term potentiation after stress exposure. Therefore, higher postsynaptic 5-HT1AR activation might predispose to a high anxious phenotype and an impaired stress coping behavior.
Asunto(s)
Hipocampo/fisiopatología , Sistema Hipotálamo-Hipofisario/fisiopatología , Potenciación a Largo Plazo/fisiología , Proteínas del Tejido Nervioso/biosíntesis , Receptor de Serotonina 5-HT1A/fisiología , Estrés Psicológico/fisiopatología , 8-Hidroxi-2-(di-n-propilamino)tetralin/farmacología , Adaptación Psicológica/efectos de los fármacos , Adaptación Psicológica/fisiología , Animales , Ansiedad/etiología , Ansiedad/fisiopatología , Corticosterona/metabolismo , Conducta Exploratoria/efectos de los fármacos , Conducta Exploratoria/fisiología , Conducta Alimentaria/efectos de los fármacos , Conducta Alimentaria/fisiología , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/fisiología , Guanosina 5'-O-(3-Tiotrifosfato)/metabolismo , Hipocampo/efectos de los fármacos , Sistema Hipotálamo-Hipofisario/efectos de los fármacos , Potenciación a Largo Plazo/efectos de los fármacos , Masculino , Ratones , Ratones Transgénicos , Proteínas del Tejido Nervioso/genética , Corteza Prefrontal/metabolismo , Proteínas Proto-Oncogénicas c-fos/biosíntesis , Proteínas Proto-Oncogénicas c-fos/genética , Receptor de Serotonina 5-HT1A/biosíntesis , Receptor de Serotonina 5-HT1A/genética , Receptores de Glucocorticoides/biosíntesis , Receptores de Glucocorticoides/genética , Receptores de Mineralocorticoides/biosíntesis , Receptores de Mineralocorticoides/genética , Proteínas Recombinantes/metabolismo , Agonistas del Receptor de Serotonina 5-HT1/farmacología , Estrés Psicológico/complicacionesRESUMEN
OBJECTIVES: Vascular mineralocorticoid receptors play a role in vascular tone and blood pressure regulation, might participate in the pathophysiology of circulatory failure during sepsis, and represent a potential therapeutic target in this disease. We aimed to study the effects of mineralocorticoids and the involvement of vascular mineralocorticoid receptors in murine endotoxic and human septic shock. DESIGN: Experimental study. SETTING: Translational investigation including animal research and in vitro experiments using human vascular cells and plasma from septic patients. SUBJECTS: Adult male C57Black 6 mice, adult patients with septic shock. INTERVENTIONS: Mice were injected with lipopolysaccharide and/or aldosterone. Human endothelial and smooth muscle cells were treated with pro-inflammatory cytokines with or without aldosterone, nuclear factor-κB inhibitor BAY 11-7082, or plasma from septic patients. MEASUREMENTS AND MAIN RESULTS: Aldosterone improved 5-day survival, invasive arterial pressure, and in vivo and ex vivo arterial response to phenylephrine at 18 hours after induction of murine endotoxic shock. Both α1-adrenoceptor and mineralocorticoid receptor expressions studied in mouse aortas were down-regulated at 6 and 18 hours in endotoxemic mice and restored in aldosterone-treated mice. Furthermore, tumor necrosis factor-α decreased both mineralocorticoid receptor and α1-adrenoceptor expressions within 5 hours in human vascular cells in a nuclear factor-κB pathway-dependent manner. Mineralocorticoid receptor expression was also blunted in human cells treated with plasma from septic patients. CONCLUSION: We found a beneficial effect of mineralocorticoids on survival, blood pressure, and vascular reactivity, associated with a restoration of α1-adrenoceptor expression in endotoxic shock. Furthermore, blunted vascular mineralocorticoid receptor expression might participate in hemodynamic failure during sepsis.
Asunto(s)
Aldosterona/farmacología , Nitrilos/farmacología , Receptores de Mineralocorticoides/biosíntesis , Choque Séptico/tratamiento farmacológico , Choque Séptico/fisiopatología , Sulfonas/farmacología , Animales , Presión Sanguínea , Citocinas/farmacología , Modelos Animales de Enfermedad , Regulación hacia Abajo , Endotoxinas , Humanos , Mediadores de Inflamación/farmacología , Lipopolisacáridos , Masculino , Ratones , Ratones Endogámicos C57BL , FN-kappa B/antagonistas & inhibidores , Choque Séptico/mortalidadRESUMEN
BACKGROUND: We have previously shown that dietary iron restriction prevents the development of renal damage in a rat model of chronic kidney disease (CKD). However, iron deficiency is associated with appetite loss. In addition, calorie restriction is reported to prevent the development of end-stage renal pathology in CKD rats. Thus, the beneficial effect of iron restriction on renal damage may depend on calorie restriction. Here, we investigate the effect of pair-feeding iron restriction on renal damage in a rat model of CKD. METHODS: First, to determine the amount of food intake, Sprague-Dawley (SD) rats were randomly given an ad libitum normal diet or an iron-restricted diet, and the food intake was measured. Second, CKD was induced by a 5/6 nephrectomy in SD rats, and CKD rats were given either a pair-feeding normal or iron-restricted diet. RESULTS: Food intake was reduced in the iron-restricted diet group compared to the normal diet group of SD rats for 16 weeks (mean food intake; normal diet group and iron-restricted diet group: 25 and 20 g/day, respectively). Based on the initial experiments, CKD rats received either a pair-feeding normal or iron-restricted diet (20 g/day) for 16 weeks. Importantly, pair-feeding iron restriction prevented the development of proteinuria, glomerulosclerosis, and tubulointerstitial damage in CKD rats. Interestingly, pair-feeding iron restriction attenuated renal expression of nuclear mineralocorticoid receptor in CKD rats. CONCLUSIONS: Pair-feeding iron restriction affected renal damage in a rat model of CKD.
Asunto(s)
Restricción Calórica , Ingestión de Alimentos , Deficiencias de Hierro , Riñón , Receptores de Mineralocorticoides/biosíntesis , Insuficiencia Renal Crónica , Animales , Modelos Animales de Enfermedad , Regulación de la Expresión Génica , Riñón/lesiones , Riñón/metabolismo , Riñón/fisiopatología , Masculino , Ratas , Ratas Sprague-Dawley , Insuficiencia Renal Crónica/metabolismo , Insuficiencia Renal Crónica/patología , Insuficiencia Renal Crónica/fisiopatologíaRESUMEN
Aldosterone is the most known mineralocorticoid hormone synthesized by the adrenal cortex. The genomic pathway displayed by aldosterone is attributed to the mineralocorticoid receptor (MR) signaling. Even though the rapid effects displayed by aldosterone are long known, our knowledge regarding the receptor responsible for such event is still poor. It is intense that the debate whether the MR or another receptor-the "unknown receptor"-is the receptor responsible for the rapid effects of aldosterone. Recently, G protein-coupled estrogen receptor-1 (GPER-1) was elegantly shown to mediate some aldosterone-induced rapid effects in several tissues, a fact that strongly places GPER-1 as the unknown receptor. It has also been suggested that angiotensin receptor type 1 (AT1) also participates in the aldosterone-induced rapid effects. Despite this open question, the relevance of the beneficial effects of aldosterone is clear in the kidneys, colon, and CNS as aldosterone controls the important water reabsorption process; on the other hand, detrimental effects displayed by aldosterone have been reported in the cardiovascular system and in the kidneys. In this line, the MR antagonists are well-known drugs that display beneficial effects in patients with heart failure and hypertension; it has been proposed that MR antagonists could also play an important role in vascular disease, obesity, obesity-related hypertension, and metabolic syndrome. Taken altogether, our goal here was to (1) bring a historical perspective of both genomic and rapid effects of aldosterone in several tissues, and the receptors and signaling pathways involved in such processes; and (2) critically address the controversial points within the literature as regarding which receptor participates in the rapid pathway display by aldosterone.
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Aldosterona/fisiología , Sistema Cardiovascular/efectos de los fármacos , Regulación de la Expresión Génica , Genómica/métodos , Insuficiencia Cardíaca , Antagonistas de Receptores de Mineralocorticoides/uso terapéutico , Receptores de Mineralocorticoides/genética , Animales , Sistema Cardiovascular/metabolismo , Insuficiencia Cardíaca/tratamiento farmacológico , Insuficiencia Cardíaca/genética , Insuficiencia Cardíaca/metabolismo , Humanos , ARN/genética , Receptores de Estrógenos/genética , Receptores de Estrógenos/metabolismo , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/metabolismo , Receptores de Mineralocorticoides/biosíntesis , Transducción de SeñalRESUMEN
BACKGROUND: Nesfatin-1, a recently identified satiety molecule derived from nucleobindin 2 (NUCB2), is associated with visceral hypersensitivity in rats and is expressed in the amygdala. We tested the hypothesis that nesfatin-1 expression in the amygdala is involved in the pathogenesis of irritable bowel syndrome (IBS) visceral hypersensitivity. METHODS: An animal model of IBS-like visceral hypersensitivity was established using maternal separation (MS) during postnatal days 2-16. The role of nesfatin-1 in the amygdala on visceral sensitivity was evaluated. KEY RESULTS: Rats subjected to MS showed a significantly increased mean abdominal withdrawal reflex (AWR) score and electromyographic (EMG) activity at 40, 60, and 80 mmHg colorectal distension. Plasma concentrations of nesfatin-1 and corticosterone were significantly higher than in non-handled (NH) rats. mRNA and protein expression of nesfatin-1/NUCB2 in the amygdala were increased in MS rats, but not in NH rats. In MS rats, AWR scores and EMG activity were significantly decreased after anti-nesfatin-1/NUCB2 injection. In normal rats, mean AWR score, EMG activity, and corticosterone expression were significantly increased after nesfatin-1 injection into the amygdala. Nesfatin-1-induced visceral hypersensitivity was abolished following application of glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) antagonists. CONCLUSIONS & INFERENCES: Elevated expression of nesfatin-1/NUCB2 in the amygdala in MS rats suggests a potential role in the pathogenesis of visceral hypersensitivity, which could potentially take place via activation of GR and MR signaling pathways.
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Amígdala del Cerebelo/metabolismo , Proteínas de Unión al Calcio/biosíntesis , Proteínas de Unión al ADN/biosíntesis , Privación Materna , Proteínas del Tejido Nervioso/biosíntesis , Receptores de Glucocorticoides/biosíntesis , Receptores de Mineralocorticoides/biosíntesis , Dolor Visceral/metabolismo , Amígdala del Cerebelo/efectos de los fármacos , Animales , Proteínas de Unión al Calcio/administración & dosificación , Proteínas de Unión al ADN/administración & dosificación , Femenino , Inyecciones Intraventriculares , Síndrome del Colon Irritable/inducido químicamente , Síndrome del Colon Irritable/metabolismo , Síndrome del Colon Irritable/fisiopatología , Masculino , Proteínas del Tejido Nervioso/administración & dosificación , Nucleobindinas , Ratas , Ratas Sprague-Dawley , Dolor Visceral/inducido químicamente , Dolor Visceral/fisiopatologíaRESUMEN
Aldosterone acts on its target tissue through a classical mechanism or through the rapid pathway through a putative membrane-bound receptor. Our goal here was to better understand the molecular and biochemical rapid mechanisms responsible for aldosterone-induced cardiomyocyte hypertrophy. We have evaluated the hypertrophic process through the levels of ANP, which was confirmed by the analysis of the superficial area of cardiomyocytes. Aldosterone increased the levels of ANP and the cellular area of the cardiomyocytes; spironolactone reduced the aldosterone-increased ANP level and cellular area of cardiomyocytes. Aldosterone or spironolactone alone did not increase the level of cyclic 3',5'-adenosine monophosphate (cAMP), but aldosterone plus spironolactone led to increased cAMP level; the treatment with aldosterone + spironolactone + BAPTA-AM reduced the levels of cAMP. These data suggest that aldosterone-induced cAMP increase is independent of mineralocorticoid receptor (MR) and dependent on Ca(2+). Next, we have evaluated the role of A-kinase anchor proteins (AKAP) in the aldosterone-induced hypertrophic response. We have found that St-Ht31 (AKAP inhibitor) reduced the increased level of ANP which was induced by aldosterone; in addition, we have found an increase on protein kinase C (PKC) and extracellular signal-regulated kinase 5 (ERK5) activity when cells were treated with aldosterone alone, spironolactone alone and with a combination of both. Our data suggest that PKC could be responsible for ERK5 aldosterone-induced phosphorylation. Our study suggests that the aldosterone through its rapid effects promotes a hypertrophic response in cardiomyocytes that is controlled by an AKAP, being dependent on ERK5 and PKC, but not on cAMP/cAMP-dependent protein kinase signaling pathways. Lastly, we provide evidence that the targeting of AKAPs could be relevant in patients with aldosterone-induced cardiac hypertrophy and heart failure.
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Proteínas de Anclaje a la Quinasa A/metabolismo , Aldosterona/administración & dosificación , Insuficiencia Cardíaca/tratamiento farmacológico , Hipertrofia/tratamiento farmacológico , Receptores de Mineralocorticoides/biosíntesis , Proteínas de Anclaje a la Quinasa A/genética , Animales , Factor Natriurético Atrial/biosíntesis , Factor Natriurético Atrial/metabolismo , AMP Cíclico/metabolismo , Ácido Egtácico/administración & dosificación , Ácido Egtácico/análogos & derivados , Insuficiencia Cardíaca/metabolismo , Humanos , Hipertrofia/metabolismo , Proteína Quinasa 7 Activada por Mitógenos/biosíntesis , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Fosforilación , Cultivo Primario de Células , Proteína Quinasa C/biosíntesis , Ratas , Receptores de Mineralocorticoides/genética , Transducción de Señal/efectos de los fármacos , Espironolactona/administración & dosificaciónRESUMEN
BACKGROUND: Ethanol (EtOH) affects the immune system. Binge drinking of hard liquor initiates a stress response. This form of drinking is popular during adolescence, which involves maturation of the immune system. The spleen is a key immune organ, and spleen atrophy is associated with immunosuppression. While the hypothalamic-pituitary-adrenal (HPA) axis plays a key role in the initial stress response, the hippocampus may be involved in stress beyond the HPA axis. METHODS: Blood ethanol concentration (BEC), blood endotoxin levels, and plasma corticosterone levels were measured following binge EtOH treatment. Absolute and relative spleen sizes were analyzed, and stress-related gene expression was compared in the hypothalamus and hippocampus. Polymerase chain reaction array was performed to analyze the expression profile of EtOH metabolism and immune regulation-related genes in the spleen. Relationships among variables were analyzed using the Pearson correlation. RESULTS: At 24 hours following a 3-day EtOH treatment, no significant difference in BEC was detected between EtOH-treated and control rats. Average plasma endotoxin levels in EtOH-treated animals were significantly higher than in controls, and spleen size was significantly lower. Spleen size did not correlate with plasma endotoxin levels; however, it did significantly negatively correlate with plasma corticosterone levels. Spleen size significantly negatively correlated with hippocampal CRH expression and significantly positively correlated with hippocampal MR expression. No correlation was observed in the hypothalamus. Significantly higher hippocampal CRH and significantly lower MR expression was seen in low spleen/body weight (sp-wt) ratio rats. No gene was found to decrease expression ≥1.5-fold (p < 0.05) in the spleen of high sp-wt group, whereas expression of several genes, including Gabra1, Gabra5, Ifnb1, Irf9, Il12b, and Cx3cr1, decreased significantly in the low sp-wt group. CONCLUSIONS: Our findings suggest that binge EtOH exposure causes lower spleen size in adolescents and that the hippocampus and stress may be associated with alterations in spleen structure and gene expression.
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Atrofia/patología , Consumo Excesivo de Bebidas Alcohólicas/patología , Etanol/efectos adversos , Hipocampo/metabolismo , Bazo/patología , Animales , Consumo Excesivo de Bebidas Alcohólicas/sangre , Corticosterona/sangre , Hormona Liberadora de Corticotropina/biosíntesis , Endotoxinas/sangre , Etanol/sangre , Expresión Génica/efectos de los fármacos , Hipotálamo/metabolismo , Masculino , Ratas , Receptores de Mineralocorticoides/biosíntesisRESUMEN
Emotional stress plays a significant role in the processes of the development of arterial hypertension, especially in the presence of genetic predisposition. The origin and maintenance of hypertensive status during stress development can be activated by the sympathetic nervous system. An increase in sympathetic stimulation can, in turn, result in a change in the functions of kidneys, which provide fluid and electrolyte balance of the organism. A comparative study of the mRNA expression level of catechol-o-methyltransferase (Comt), mineralocorticoid receptor (Mlr), and ß-subunit of epithelial sodium channel (ß-ENaC) genes was conducted on the kidneys of hypertensive ISIAH rats and normotensive WAG rats at rest and after the effect of emotional stress. The discovered changes in the expression level of the selected genes confirm their involvement in increased sympathetic stimulation of the kidney, along with changes in the function of kidney regulation of fluid and electrolyte balance, which is an important factor of the development of sustained hypertension in the ISIAH rats strain.
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Catecol O-Metiltransferasa/biosíntesis , Canales Epiteliales de Sodio/biosíntesis , Hipertensión/genética , Receptores de Mineralocorticoides/biosíntesis , Animales , Catecol O-Metiltransferasa/genética , Canales Epiteliales de Sodio/genética , Regulación de la Expresión Génica , Predisposición Genética a la Enfermedad , Humanos , Hipertensión/patología , Riñón/metabolismo , Riñón/patología , ARN Mensajero/biosíntesis , Ratas , Receptores de Mineralocorticoides/genética , Estrés Psicológico/genéticaRESUMEN
CONTEXT: Hypertension is one of the most important risk factors in cardiovascular disease and is the most common chronic disease. Mineralocorticoid receptor (MR) antagonists have been successfully used in clinic for the treatment of hypertension. OBJECTIVE: Our study aims to investigate whether Arctigenin can antagonize MR and inhibit the transcription of Na/K-ATPase. MATERIALS AND METHODS: The yeast two-hybrid assay was used to screen natural products and Arctigenin was identified as an MR antagonist. The direct binding of Arctigenin to MR was determined using assays based on surface plasmon resonance, differential scanning calorimetry and fluorescence quenching. Furthermore, results from mammalian one-hybrid and transcriptional activation experiments also confirmed that Arctigenin can potently antagonize MR in cells. We demonstrated that Arctigenin can decrease the level of Na/K-ATPase mRNA by antagonizing MR in HK-2 cells. RESULTS: Our findings show that Arctigenin can effectively decrease Na/K-ATPase transcription; thus highlight its potential as an anti-hypertensive drug lead compound. DISCUSSION AND CONCLUSION: Our current findings demonstrate that Arctigenin is an antagonist of MR and effectively decreases the Na/K-ATPase 1 gene expression. Our work provides a hint for the drug discovery against cardiovascular disease.
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Furanos/administración & dosificación , Hipertensión/tratamiento farmacológico , Lignanos/administración & dosificación , Antagonistas de Receptores de Mineralocorticoides/administración & dosificación , Receptores de Mineralocorticoides/biosíntesis , ATPasa Intercambiadora de Sodio-Potasio/genética , Animales , Regulación de la Expresión Génica/efectos de los fármacos , Células HEK293 , Humanos , Hipertensión/genética , Hipertensión/patología , ARN Mensajero/biosíntesis , ATPasa Intercambiadora de Sodio-Potasio/biosíntesis , Resonancia por Plasmón de Superficie , Activación Transcripcional/efectos de los fármacosRESUMEN
OBJECTIVE: To explore the role of glucocorticoid new mechanism to observe the expression of glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) with lipopolysaccharide (LPS) and dexamethasone (Dex) in human umbilical vein endothelial cells (HUVEC). MATERIALS AND METHODS: LPS "injured" endothelial cells with Dex for "treatment", and then detected the expression of glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) in the endothelial cells by RT-PCR and immunohistochemistry. RESULTS: With high dose (10-6 mol/L) of Dex to stimulate cell 3h, GRmRNA no significant changes in the expression, 6h began to decrease, 12h peak, 24h recovered nearly the level before stimulation. Using different concentrations of Dex and 100 ng/ml LPS stimulation, HUVEC MRmRNA expression was decreased, and high dose (10-6 mol/L) of Dex to stimulate cell 3h, MRmRNA no significant changes in expression, and GRmRNA The difference is that the expression began to increase 6h, 12h, peaked, 24h rebound near the level before stimulation. Immunohistochemistry results consistent with the RT-PCR. CONCLUSIONS: Large dose of DEX (10-6 mol/l) up-regulated the expression of MR and GR in the reduction of the contrast exactly. GC induced the expression of GR and MR in different changes of stress injury of the body may be a regulatory mechanism, and indicate one new mechanism of glucocorticoid exist.