RESUMEN
The aim of the study was to elucidate the involvement of cholinergic receptor nicotinic alpha 1 subunit (CHRNA1) in the pathogenesis of primary focal hyperhidrosis (PFH). The hyperhidrosis mouse model was constructed using pilocarpine injection. The expression levels of CHRNA1 in sweat gland tissues of PFH patients and hyperhidrosis mice were compared using Western blots and quantitative real-time PCR (qRT-PCR) analyses. Sweat secretion in hyperhidrosis mice treated with small-interfering RNA (siRNA) targeting CHRNA1 (si-CHRNA1) or non-specific siRNA were compared. Sweat secretory granules in the sweat gland cells of hyperhidrosis mice were examined using transmission electron microscopy. The serum level of acetylcholine was measured using enzyme-linked immunosorbent assay, while markers associated with PFH, including Aquaporin 5 (AQP5) and Calcium Voltage-Gated Channel Subunit Alpha1 C (CACNA1C), were assessed using immunohistochemical assay and Western blots. Brain-derived neurotrophic factor (BDNF) and Neuregulin 1 (NRG-1) in sympathetic ganglia axons of hyperhidrosis mice were quantified using Western blots. CHRNA1 up-regulation is a characteristic of the sweat glands of PFH patients and Hyperhidrosis mice. Silencing CHRNA1 decreased sweat secretion and the number of sweat secretory granules of hyperhidrosis mice. Serum acetylcholine, as well as AQP5 and CACNA1C expression in the sweat glands, was reduced by siCHRNA1. BDNF1 and NRG-1 levels in the sympathetic ganglia axons were also attenuated by siCHRNA1 treatment. CHRNA1 up-regulation is a potential biomarker of PFH and downregulating CHRNA1 could alleviate the symptoms of PFH through inactivating the sympathetic system.
Asunto(s)
Hiperhidrosis/metabolismo , Receptores Nicotínicos/metabolismo , Glándulas Sudoríparas/metabolismo , Acetilcolina/sangre , Animales , Acuaporina 5/genética , Acuaporina 5/metabolismo , Canales de Calcio Tipo L/genética , Canales de Calcio Tipo L/metabolismo , Humanos , Hiperhidrosis/genética , Ratones , Ratones Endogámicos BALB C , Receptores Nicotínicos/genéticaRESUMEN
The aim was to investigate the role of the α7nAChR-mediated cholinergic anti-inflammatory pathway in vagal nerve regulated atrial fibrillation (AF).18 beagles (standard dogs for testing) were used in this study, and the effective refractory period (ERP) of atrium and pulmonary veins and AF inducibility were measured hourly during rapid atrial pacing at 800 beats/minute for 6 hours in all beagles. After cessation of 3 hours of RAP, the low-level vagal nerve stimulation (LL-VNS) group (n = 6) was given LL-VNS and injection of salinne (0.5 mL/GP) into four GPs, the methyllycaconitine (MLA, the antagonist of α7nAChR) group (n = 6) was given LL-VNS and injection of MLA into four GPs, and the Control group (n = 6) was given saline into four GPs and the right cervical vagal nerve was exposed without stimulation. Then, the levels of the tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), acetylcholine (ACh), STAT3, and NF-κB proteins were measured. During the first 3 hours of RAP, the ERPs gradually decreased while the dispersion of ERPs (dERPs) and AF inducibility gradually increased in all three groups. During the last 3 hours of 6 hours' RAP in this study, the ERPs in the LL-VNS group were higher, while the dERPs and AF inducibility were significantly lower when compared with the Control and MLA groups at the same time points. The levels of ACh in the serum and atrium in the LL-VNS and MLA groups were higher than in the Control group, and the levels of TNF-α and IL-6 were higher in the Control and MLA groups than in the LL-VNS group. The concentrations of STAT3 in RA and LA tissues were higher in the LL-VNS group while those of NF-κB were lower.In conclusion, the cholinergic anti-inflammatory pathway mediated by α7nACh plays an important role in low-level vagal nerve-regulated AF.
Asunto(s)
Aconitina/análogos & derivados , Fibrilación Atrial/fisiopatología , Neuroinmunomodulación/efectos de los fármacos , Nervio Vago/efectos de los fármacos , Receptor Nicotínico de Acetilcolina alfa 7/antagonistas & inhibidores , Acetilcolina/sangre , Aconitina/administración & dosificación , Aconitina/farmacología , Animales , Estimulación Cardíaca Artificial/efectos adversos , Estimulación Cardíaca Artificial/métodos , Estudios de Casos y Controles , Modelos Animales de Enfermedad , Perros , Atrios Cardíacos/inervación , Atrios Cardíacos/fisiopatología , Interleucina-6/sangre , FN-kappa B/sangre , Antagonistas Nicotínicos/administración & dosificación , Antagonistas Nicotínicos/farmacología , Venas Pulmonares/inervación , Venas Pulmonares/fisiopatología , Periodo Refractario Electrofisiológico/efectos de los fármacos , Factor de Transcripción STAT3/sangre , Factor de Necrosis Tumoral alfa/sangre , Estimulación del Nervio Vago/efectos adversos , Estimulación del Nervio Vago/métodosRESUMEN
BACKGROUND: Organ ischemia-reperfusion injury often induces local and systemic inflammatory responses, which in turn worsen organ injury. These inflammatory responses can be regulated by the central nervous system, particularly by the vagal nerve and nicotinic acetylcholine receptors, which are the key components of cholinergic anti-inflammatory pathway. Activation of the cholinergic anti-inflammatory pathway can suppress excessive inflammatory responses and be a potential strategy for prevention of ischemia-reperfusion injury of organs including the kidney. METHODS: Vagal nerve activity, plasma acetylcholine, catecholamine and inflammatory mediators, renal tissue injury, and cell death were measured in mice with bilateral renal ischemia/reperfusion with or without treatment with dexmedetomidine (Dex), an α2-adrenergic receptor agonist. RESULTS: Dex significantly increased the discharge frequency of the cervical vagal nerve by up to 142 Hz (mean) (P < .001), and preserved kidney gross morphology and structure and attenuated cell apoptosis after ischemia-reperfusion. Furthermore, Dex also significantly increased acetylcholine release to 135.8 pmol/L (median) when compared to that (84.7 pmol/L) in the sham group (P < .001) and reduced the levels of several inflammatory mediators induced by renal ischemia/reperfusion. All the effects were abolished by vagotomy, splenectomy, or combinative administration of atipamezole, an α2-adrenergic receptor antagonist. CONCLUSIONS: Our findings suggest that Dex provides renoprotection, at least in part, through anti-inflammatory effects of the parasympathetic nervous system activation in addition to its direct actions on α2-adrenergic receptors.
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Agonistas de Receptores Adrenérgicos alfa 2/farmacología , Agonistas de Receptores Adrenérgicos alfa 2/uso terapéutico , Antiinflamatorios/farmacología , Dexmedetomidina/farmacología , Dexmedetomidina/uso terapéutico , Enfermedades Renales/prevención & control , Sistema Nervioso Parasimpático/efectos de los fármacos , Daño por Reperfusión/prevención & control , Acetilcolina/sangre , Antagonistas de Receptores Adrenérgicos alfa 2/farmacología , Animales , Apoptosis/efectos de los fármacos , Catecolaminas/sangre , Imidazoles/farmacología , Mediadores de Inflamación/metabolismo , Riñón/patología , Enfermedades Renales/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Nervio Vago/fisiopatologíaRESUMEN
New reliable and robust potentiometric ion-selective electrodes were fabricated using poly(3,4-ethylenedioxythiophene)/poly(styrenesulfonate) (PEDOT/PSS) as the solid contact between the sensing membrane and electrical substrate for an acetylcholine (ACh) bioassay. A film of PEDOT/PSS was deposited on a solid carbon screen-printed platform made from ceramic substrate. The selective materials used in the ion-selective electrode (ISE) sensor membrane were acetylcholinium tetraphenylborate (ACh/TPB/PEDOT/PSS-ISE) (sensor I) and triacetyl-ß-cyclodextrin (ß-CD/PEDOT/PSS-ISE) (sensor II). The sensors revealed clear enhanced Nernstian response with a cationic slope 56.4 ± 0.6 and 55.3 ± 1.1 mV/decade toward (ACh+) ions over the dynamic linear range 1.0 × 10-6-1 × 10-3 and 2.0 × 10-6-1.0 × 10-3 M at pH 5 with limits of detection 2.0 × 10-7 and 3.2 × 10-7 M for sensors I and II, respectively. The selectivity behavior of both sensors was also tested and the sensors showed a significant high selectivity toward ACh+ over different common organic and inorganic cations. The stability of the potential response for the solid-contact (SC)/ISEs was evaluated using a chronopotentiometric method and compared with that of electrodes prepared without adding the solid-contact material (PEDOT/PSS). Enhanced accuracy, excellent repeatability, good reproducibility, potential stability, and high selectivity and sensitivity were introduced by these cost-effective sensors. The sensors were also used to measure the activity of acetylcholinesterase (AChE). A linear plot between the initial rate of the hydrolysis of ACh+ substrate and enzyme activity held 5.0 × 10-3-5.2 IUâL-1 of AChE enzyme. Application to acetylcholine determination in human serum was done and the results were compared with the standard colorimetric method.
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Acetilcolina/sangre , Acetilcolinesterasa/sangre , Técnicas Biosensibles/métodos , Compuestos Bicíclicos Heterocíclicos con Puentes/química , Carbono/química , Neurotransmisores/sangre , Polímeros/química , Poliestirenos/química , Impresión , Análisis de Inyección de Flujo , Humanos , Hidrodinámica , Hidrólisis , Cinética , Potenciometría , Agua/químicaRESUMEN
BACKGROUND/AIMS: Vagus nerve stimulation (VNS) suppresses arrhythmic activity and minimizes cardiomyocyte injury. However, how VNS affects angiogenesis/arteriogenesis in infarcted hearts, is poorly understood. METHODS: Myocardial infarction (MI) was achieved by ligation of the left anterior descending coronary artery (LAD) in rats. 7 days after LAD, stainless-steel wires were looped around the left and right vagal nerve in the neck for vagus nerve stimulation (VNS). The vagal nerve was stimulated with regular pulses of 0.2ms duration at 20 Hz for 10 seconds every minute for 4 hours, and then ACh levels by ELISA in cardiac tissue and serum were evaluated for its release after VNS. Three and 14 days after VNS, Real-time PCR, immunostaining and western blot were respectively used to determine VEGF-A/B expressions and α-SMA- and CD31-postive vessels in VNS-hearts with pretreatment of α7-nAChR blocker mecamylamine (10 mg/kg, ip) or mACh-R blocker atropine (10 mg/kg, ip) for 1 hour. The coronary function and left ventricular performance were analyzed by Langendorff system and hemodynamic parameters in VNS-hearts with pretreatment of VEGF-A/B-knockdown or VEGFR blocker AMG706. Coronary arterial endothelial cells proliferation, migration and tube formation were evaluated for angiogenesis following the stimulation of VNS in coronary arterial smooth muscle cells (VSMCs). RESULTS: VNS has been shown to stimulate VEGF-A and VEGF-B expressions in coronary arterial smooth muscle cells (VSMCs) and endothelial cells (ECs) with an increase of α-SMA- and CD31-postive vessel number in infarcted hearts. The VNS-induced VEGF-A/B expressions and angiogenesis were abolished by m-AChR inhibitor atropine and α7-nAChR blocker mecamylamine in vivo. Interestingly, knockdown of VEGF-A by shRNA mainly reduced VNS-mediated formation of CD31+ microvessels. In contrast, knockdown of VEGF-B powerfully abrogated VNS-induced formation of α-SMA+ vessels. Consistently, VNS-induced VEGF-A showed a greater effect on EC tube formation as compared to VNS-induced VEGF-B. Moreover, VEGF-A promoted EC proliferation and VSMC migration while VEGF-B induced VSMC proliferation and EC migration in vitro. Mechanistically, vagal neurotransmitter acetylcholine stimulated VEGF-A/B expressions through m/nACh-R/PI3K/Akt/Sp1 pathway in EC. Functionally, VNS improved the coronary function and left ventricular performance. However, blockade of VEGF receptor by antagonist AMG706 or knockdown of VEGF-A or VEGF-B by shRNA significantly diminished the beneficial effects of VNS on ventricular performance. CONCLUSION: VNS promoted angiogenesis/arteriogenesis to repair the infracted heart through the synergistic effects of VEGF-A and VEGF-B.
Asunto(s)
Infarto del Miocardio/terapia , Estimulación del Nervio Vago , Factor A de Crecimiento Endotelial Vascular/metabolismo , Factor B de Crecimiento Endotelial Vascular/metabolismo , Acetilcolina/análisis , Acetilcolina/sangre , Animales , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Indoles/farmacología , Masculino , Microvasos/citología , Microvasos/efectos de los fármacos , Microvasos/metabolismo , Músculo Liso Vascular/citología , Músculo Liso Vascular/efectos de los fármacos , Músculo Liso Vascular/metabolismo , Infarto del Miocardio/patología , Miocardio/metabolismo , Niacinamida/administración & dosificación , Niacinamida/farmacología , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/metabolismo , Interferencia de ARN , ARN Interferente Pequeño/metabolismo , Ratas , Ratas Sprague-Dawley , Receptores Muscarínicos/química , Receptores Muscarínicos/metabolismo , Factor A de Crecimiento Endotelial Vascular/antagonistas & inhibidores , Factor A de Crecimiento Endotelial Vascular/genética , Factor B de Crecimiento Endotelial Vascular/antagonistas & inhibidores , Factor B de Crecimiento Endotelial Vascular/genética , Receptor Nicotínico de Acetilcolina alfa 7/antagonistas & inhibidores , Receptor Nicotínico de Acetilcolina alfa 7/metabolismoRESUMEN
This study was undertaken to assess the influence of an Anaplasma marginale infection on oxidative stress and antioxidant status, trace elements and cholinesterase as markers of the inflammatory process and biomarkers of oxidative imbalance. An infected group comprised of 35 crossbred Holstein cattle, about 2-3 years old, naturally infected with Anaplasma marginale, were divided into 4 subgroups according to their parasitemia rates (<1%, 1-10%, 10-20%, >20%) and also 10 healthy cattle as control were selected. Blood samples were taken and hematological parameters, activities of antioxidant enzymes including erythrocyte glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), catalase (CAT), glucose-6-phosphate dehydrogenase (G6PD), total antioxidant capacity (TAC), median corpuscularfragility (MCF) as well as acetylcholinesterase (AChE), and serum concentrations of antioxidant trace minerals (copper, iron, zinc, manganese, and selenium) and butyrylcholinesterase (BChE) were determined. In addition, as an index of lipid peroxidation, the level of malondialdehyde (MDA) was measured. The results revealed a significant decrease (Pâ¯<â¯0.05) in RBC count, packed cell volume (PCV) and Hb concentration as well as the activities of erythrocyte GSH-Px, SOD, CAT, G6PD, TAC, MCF and AChE and serum concentrations of Cu, Zn, Mn, Se and BchE in the infected cattle. In contrast, significantly increased (Pâ¯<â¯0.05) levels of MDA and erythrocyte osmotic fragility as well as serum concentration of iron were recorded in the infected animals. The significant decrease in antioxidant enzyme activities and substantial elevated levels of lipid peroxidation and erythrocyte osmotic fragility associated with the notable increase in parasitemia indicate increased exposure of RBCs to oxidative damage. Furthermore, decrease of cholinesterase in infection by A. marginale can and directly or indirectly lead to increase acetylcholine levels potent anti-inflammatory molecules, thereby inhibiting inflammation.
Asunto(s)
Anaplasma marginale/patogenicidad , Anaplasmosis/sangre , Antioxidantes , Biomarcadores , Enfermedades de los Bovinos/sangre , Colinesterasas/metabolismo , Estrés Oxidativo , Oligoelementos/sangre , Acetilcolina/sangre , Acetilcolinesterasa/sangre , Anaplasmosis/epidemiología , Anemia , Animales , Butirilcolinesterasa/sangre , Catalasa/sangre , Bovinos , Enfermedades de los Bovinos/diagnóstico , Enfermedades de los Bovinos/epidemiología , Recuento de Eritrocitos , Eritrocitos/metabolismo , Glucosafosfato Deshidrogenasa/sangre , Glutatión Peroxidasa/sangre , Irán/epidemiología , Peroxidación de Lípido , Malondialdehído/sangre , Fragilidad Osmótica , Parasitemia/sangre , Suero/química , Suero/enzimología , Superóxido Dismutasa/sangreRESUMEN
Chagas disease is an acute or chronic illness that causes severe inflammatory response, and consequently, it may activate the inflammatory cholinergic pathway, which is regulated by cholinesterases, including the acetylcholinesterase. This enzyme is responsible for the regulation of acetylcholine levels, an anti-inflammatory molecule linked to the inflammatory response during parasitic diseases. Thus, the aim of this study was to investigate whether Trypanosoma cruzi infection can alter the activity of acetylcholinesterase and acetylcholine levels in mice, and whether these alterations are linked to the inflammatory cholinergic signaling pathway. Twenty-four mice were divided into two groups: uninfected (control group, n = 12) and infected by T. cruzi, Y strain (n = 12). The animals developed acute disease with a peak of parasitemia on day 7 post-infection (PI). Blood, lymphocytes, and brain were analyzed on days 6 and 12 post-infection. In the brain, acetylcholine and nitric oxide levels, myeloperoxidase activity, and histopathology were analyzed. In total blood and brain, acetylcholinesterase activity decreased at both times. On the other hand, acetylcholinesterase activity in lymphocytes increased on day 6 PI compared with the control group. Infection by T. cruzi increased acetylcholine and nitric oxide levels and histopathological damage in the brain of mice associated to increased myeloperoxidase activity. Therefore, an intense inflammatory response in mice with acute Chagas disease in the central nervous system caused an anti-inflammatory response by the activation of the cholinergic inflammatory pathway.
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Acetilcolina/sangre , Acetilcolinesterasa/sangre , Encéfalo/metabolismo , Cardiomiopatía Chagásica/sangre , Linfocitos/metabolismo , Trypanosoma cruzi , Animales , Encéfalo/patología , Cardiomiopatía Chagásica/patología , Linfocitos/patología , Ratones , Óxido Nítrico/sangre , Peroxidasa/sangreRESUMEN
Aims: Studies have shown that stellate ganglion nerve activity has association with atrial electrical remodelling and atrial fibrillation (AF) inducibility, while median nerve stimulation (MNS) decreases cardiac sympathetic drive. In this study, we tested the hypothesis that MNS suppresses atrial electrical remodelling and AF vulnerability. Methods and results: The atrial effective refractory period (AERP) and AF inducibility at baseline and after 3 h of rapid atrial pacing were determined in dogs undergoing MNS (n = 7), MNS+ application of methyllycaconitine (n = 7) or sham procedure (n = 6). Then, the levels of tumour necrosis factor-alpha (TNF-a), interleukin-6 (IL-6), and acetylcholine (Ach) in the plasma and atrial tissues were measured. The control dogs (n = 4) were assigned to measure atrial inflammation cytokines. Short-term rapid atrial pacing induced shortening of the AERP, an increase in AERP dispersion, and an increase AF vulnerability in the sham dogs, which were all suppressed by MNS. Levels of TNF-a and IL-6 were higher, and Ach levels were lower in the left and the right atrium in the sham dogs than in the MNS dogs. Methyllycaconitine blunted the effects of MNS on the AERP, AERP dispersion, the AF vulnerability, and TNF-a and IL-6 levels in the atrium, but had no impact on the levels of Ach. Conclusions: The effects of MNS on atrial electrical remodelling and AF inducibility might be associated with the cholinergic anti-inflammatory pathway.
Asunto(s)
Fibrilación Atrial/terapia , Remodelación Atrial , Sistema Nervioso Autónomo/fisiopatología , Estimulación Cardíaca Artificial , Terapia por Estimulación Eléctrica/métodos , Atrios Cardíacos/inervación , Frecuencia Cardíaca , Mediadores de Inflamación/sangre , Nervio Mediano , Acetilcolina/sangre , Potenciales de Acción , Animales , Fibrilación Atrial/sangre , Fibrilación Atrial/etiología , Fibrilación Atrial/fisiopatología , Sistema Nervioso Autónomo/metabolismo , Modelos Animales de Enfermedad , Perros , Interleucina-6/sangre , Periodo Refractario Electrofisiológico , Factor de Necrosis Tumoral alfa/sangreRESUMEN
PURPOSE: Individuals predisposed to vasovagal syncope may have different autonomic nervous system control mechanisms from those without predisposition to vasovagal events. To test this hypothesis, we investigated different sympathetic responses in a canine model of vasovagal syncope. METHODS: Left thoracotomy was performed on 20 mongrel dogs. The heart was exposed and a bolus of veratridine (15 µg/kg), a neurotoxin which prevents the inactivation of sodium ion channels, was injected into the left atrium to induce a Bezold-Jarisch reflex-mediated vasovagal event, characterized by bradycardia, decreased inotropism, and hypotension. Electrocardiogram and blood pressure were continuously monitored. Neural activity was recorded from the left stellate ganglion. Plasma norepinephrine and acetylcholine levels were measured 30 s before and 30 s after veratridine injection. RESULTS: Veratridine resulted in rapid decreases in heart rate and blood pressure in all dogs, accompanied by increases in both norepinephrine and acetylcholine. Two types of neural activity (high-amplitude spike discharge activity and low-amplitude burst discharge activity) were recorded from the left stellate ganglion. Veratridine induced high-frequency spike discharge activity in some dogs (Group A), whereas spike discharge activity was scarce and relatively unresponsive to veratridine in the remaining dogs (Group B). Dogs in Group A had higher plasma norepinephrine levels (111.63 ± 15.1 vs. 48.11 ± 33.81 ng/l, p = 0.002) and less intense drops in heart rate (- 37 ± 24 vs. - 84 ± 28 bpm, p = 0.001) and blood pressure (systolic blood pressure, - 18 ± 15 vs. - 37 ± 13 mmHg, p = 0.009; diastolic blood pressure, - 26 ± 13 vs. - 45 ± 13 mmHg, p = 0.005) compared to dogs in Group B. Similarly, heart rate post-veratridine was higher (102 ± 23 vs. 69 ± 22 bpm, p = 0.004), the veratridine-induced longest RR interval was shorter (0.7 [0.5-0.8] vs. 1.2 [1.1-3.5] s, p < 0.001) and the diastolic and mean arterial pressures post-veratridine were higher (all p < 0.05) in dogs in Group A compared to those in Group B. CONCLUSIONS: Distinct sympathetic activation as represented by left stellate ganglion high-frequency spike discharge activity protected against bradycardia and hypotension in a canine model of vasovagal syncope. Our findings may have therapeutic implications.
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Sistema Nervioso Simpático/fisiopatología , Síncope Vasovagal/fisiopatología , Acetilcolina/sangre , Animales , Presión Arterial , Presión Sanguínea , Perros , Electrocardiografía , Frecuencia Cardíaca , Hipotensión/fisiopatología , Masculino , Contracción Miocárdica , Norepinefrina/sangre , Bloqueadores de los Canales de Sodio , Ganglio Estrellado/fisiopatología , Síncope Vasovagal/inducido químicamente , VeratridinaRESUMEN
This study evaluated the effects of temperature on the activities of antioxidant enzymes and hormone levels in Schizothorax prenanti. Schizothorax prenanti were acutely suffered from increasing temperature at a rate of 1⯰Câ¯h-1. The temperature started from 11⯰C and the critical thermal maximum (CTMax) was measured to evaluated thermal tolerance of Schizothorax prenanti. Antioxidant parameters including superoxide dismutase (SOD), catalase (CAT), Na+/K+-ATPase and malondialdehyde (MDA) in gills, liver and muscle were measured at five temperature groups (11⯰C, 16⯰C, 21⯰C, 26⯰C and 31⯰C). The plasma hormone including acetylcholine (ACh) and cortisol were also measured at five temperature groups. Our results showed that the CTMax of Schizothorax prenanti under acute heat stress was 31⯰C.The activities of SOD and CAT in liver and muscle at 11⯰C, 16⯰C, 26⯰C and 31⯰C groups were significantly higher than 21⯰C group, as well as Na+/K+-ATPase in gills and MDA concentrations in gills and liver. However, plasma ACh and cortisol levels were significantly increased with increasing temperature. The results indicate that oxidative stress parameters could respons to increase of temperatur, altogether reflect that Schizothorax prenanti has higher susceptibility of temperature. Thus, the effect of long-term high temperature on Schizothorax prenanti should be studied further.
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Antioxidantes/metabolismo , Cyprinidae/metabolismo , Proteínas de Peces/metabolismo , Respuesta al Choque Térmico , Acetilcolina/sangre , Adaptación Fisiológica , Animales , Catalasa/metabolismo , Calor , Hidrocortisona/sangre , Malondialdehído/metabolismo , Estrés Oxidativo , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , Superóxido Dismutasa/metabolismoRESUMEN
BACKGROUND: Non-neuronal acetylcholine (ACh) restricts autoimmune responses and attenuates inflammation by cholinergic anti-inflammation pathway. To date, the implication of ACh in myasthenia gravis (MG) remained unexplored. This study aimed to investigate the possible relationship between ACh levels, anti-muscle-specific tyrosine kinase (MuSK) antibody titers, main clinical features and outcomes of MG patients. METHODS: We successfully measured ACh levels in human peripheral blood mononuclear cells (PBMCs) from 125 MG patients and 50 matched healthy controls by using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). We assessed the quantitative MG (QMG) scores for each patient and titered anti-MuSK antibody. RESULTS: We found that PBMC-derived ACh level was significantly higher in MG patients, especially in patients of class III, IV-V, compared with that in controls (0.142 ± 0.108 vs. 0.075 ± 0.014 ng/million cells, p = 0.0003) according to the Myasthenia Gravis Foundation of America clinical classification. Importantly, we also found that ACh levels were positively correlated with QMG scores (r = 0.83, p < 0.0001) and anti-MuSK Ab levels (r = 0.85, p < 0.0001). CONCLUSIONS: Our demonstration of elevated ACh levels in PBMCs of MG patients foreshadows potential new avenues for MG research and treatment.
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Acetilcolina/sangre , Leucocitos Mononucleares/metabolismo , Miastenia Gravis/sangre , Adulto , Autoanticuerpos/sangre , Femenino , Humanos , Masculino , Persona de Mediana Edad , Proteínas Tirosina Quinasas Receptoras/sangre , Receptores Colinérgicos/sangre , Espectrometría de Masas en TándemRESUMEN
Prevalence of dermatological disorder in patients with end-stage kidney disease is estimated as 50% to 100% in various studies. Some of the skin lesions are specific for the diseases causing chronic kidney disease (CKD), some are associated with CKD, and still others are the dermatological manifestation of uremia. Microangiopathy was also found in both arterioles and venule in the skin biopsy of "normal looking" skin in patients with end-stage kidney disease. In a cross-sectional study in patients on dialysis, we measured skin blood flow (SBF) using laser Doppler device in a standardized way at various areas of lower extremities at 2 different local skin temperatures: 35°C and 44°C. Local heating increases skin perfusion by mechanisms dependent on nitric oxide (NO). SBF was impaired in CKD patients Stage 5 on HD, particularly in those with diabetes mellitus as a cause of CKD. The reduced response in the SBF to the heat in our patients may be due to decreased generation of NO in uremia. Endothelium-dependent vasodilatation in patients on dialysis and the response of the skin microcirculation to acetylcholine was diminished in hypertensive patients on dialysis. Similarly, patients with diabetes mellitus had decreased SBF during intradermal microdialysis with a NO synthase inhibitor. Multiple uremic toxins have been studied in vitro and show to cause various degree of endothelial cell dysfunction. Unfortunately, no clear benefit has been described in CKD patients to different intervention aimed to reduce uremic toxin effect on endothelium. There are no long-term data on the factors which can modify endothelium function in uremia, but non pharmacologic interventions, diet, and several pharmacologic approaches could be beneficial. Measurement of SBF can be useful in evaluation of vasculopathy in CKD population and can potentially be used for assessment of vascular response during specific clinical intervention.
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Endotelio Vascular/fisiología , Enfermedades de la Piel/sangre , Piel/irrigación sanguínea , Uremia/sangre , Acetilcolina/sangre , Estudios de Casos y Controles , Humanos , Fallo Renal Crónico/sangre , Fallo Renal Crónico/complicaciones , Fallo Renal Crónico/terapia , Óxido Nítrico/sangre , Flujo Sanguíneo Regional , Diálisis Renal , Enfermedades de la Piel/complicaciones , Enfermedades de la Piel/terapia , Uremia/complicaciones , Uremia/terapiaRESUMEN
OBJECTIVES: In this work, we hypothesized whether galantamine could interact with the cholinergic anti-inflammatory pathway and modulate immunity this way. BACKGROUND: Galantamine is a drug used for the therapy of Alzheimer disease. The drug inhibits enzyme acetylcholinesterase in the central nervous system, which causes better availability of neurotransmitter acetylcholine. METHODS: In the experiment, we immunized BALB/c laboratory mice by keyhole limpet hemocyanin (KLH) in combination with galantamine in a dose 0.02-0.5 mg/kg. The animals were sacrificed from 1 to 7 days after the substances applications and plasma was collected in order to examine immunochemical markers by enzyme-linked immunosorbent assay. RESULTS: We found significant drop in production of immunoglobulins and interleukin (IL) 4 level while IL2, IL4 and tumour necrosis factor α remained unaltered for the whole experiment. We infer that galantamine causes better availability of acetylcholine also in blood system, where the neurotransmitter interacts with nicotinic acetylcholine receptors on macrophages and initiates cholinergic anti-inflammatory pathway. CONCLUSIONS: In a conclusion, galantamine can cause lower efficacy of vaccination or immunity response to an infectious disease and the phenomenon should be taken into consideration in the current therapy (Tab. 1, Fig. 2, Ref. 24).
Asunto(s)
Acetilcolinesterasa/sangre , Enfermedad de Alzheimer/inmunología , Antiinflamatorios/farmacología , Inhibidores de la Colinesterasa/sangre , Galantamina/farmacología , Hemocianinas/farmacología , Acetilcolina/sangre , Animales , Interacciones Farmacológicas , Ensayo de Inmunoadsorción Enzimática , Femenino , Inmunoglobulinas/sangre , Mediadores de Inflamación/sangre , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Ratones , Ratones Endogámicos BALB C , Receptores Nicotínicos/efectos de los fármacosRESUMEN
Atrial fibrillation (AF) is the most prevalent heart rhythm disorder, and autonomic nervous system (ANS) is important to AF. This study aims to identify whether changes in transmitters released by ANS could reflect their activities. The right atrium (RA) groups (1-40V) included RA500 and RA1000. While ANS groups received high-frequency electrical stimulation (1-8V, 20 Hz, 2 ms), including left stellate ganglion stimulation (LSGS) andleft cervical vagus trunk stimulation (LVTS). The induced rate of AF, duration and atrial effective refractory period (AERP) were measured. The blood was drawn for evaluation of norepinephrine (NE) and acetylcholine (Ach) concentrations. At 12-hours, RA tissue was dissected and compared against un-stimulated controls. While AF was induced by all groups, duration and AERP were significantly different between RA pacing groups and ANS-stimulated groups, respectively (P<0.05). Specific changes in profile of NE and Ach were associated with modality of stimulation. RA1000 tended to display most significant changes (P<0.05) compared to other groups while variables concentration levels were observed in other groups. In conclusion, electrically-induced AF initiated by various modalities of stimulation showed different changes in serum and RA tissues. Fast frequency pacing caused significant atrial electrical remodeling, including ANS activity change.
Asunto(s)
Acetilcolina/sangre , Fibrilación Atrial/sangre , Fibrilación Atrial/fisiopatología , Fenómenos Electrofisiológicos , Norepinefrina/sangre , Animales , Modelos Animales de Enfermedad , Perros , Atrios Cardíacos/patología , Atrios Cardíacos/fisiopatología , Periodo Refractario ElectrofisiológicoRESUMEN
Multiple sclerosis (MS) is characterized by pro-inflammatory cytokine production. Acetylcholine (ACh) contributes to the modulation of central and peripheral inflammation. We studied the homeostasis of the cholinergic system in relation to cytokine levels in immune cells and sera of relapsing remitting-MS (RR-MS) patients. We demonstrated that lower ACh levels in serum of RR-MS patients were inversely correlated with the increased activity of the hydrolyzing enzymes acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE). Interestingly, the expression of the ACh biosynthetic enzyme and the protein carriers involved in non-vesicular ACh release were found overexpressed in peripheral blood mononuclear cells of MS patients. The inflammatory state of the MS patients was confirmed by increased levels of TNFα, IL-12/IL-23p40, IL-18. The lower circulating ACh levels in sera of MS patients are dependent on the higher activity of cholinergic hydrolyzing enzymes. The smaller ratio of ACh to TNFα, IL-12/IL-23p40 and IL-18 in MS patients, with respect to healthy donors (HD), is indicative of an inflammatory environment probably related to the alteration of cholinergic system homeostasis.
Asunto(s)
Acetilcolina/sangre , Esclerosis Múltiple Recurrente-Remitente/sangre , Acetilcolinesterasa/metabolismo , Adulto , Anciano , Butirilcolinesterasa/metabolismo , Citocinas/sangre , Femenino , Humanos , Interleucina-12/sangre , Interleucina-18/sangre , Masculino , Persona de Mediana Edad , Esclerosis Múltiple Recurrente-Remitente/enzimología , Factor de Necrosis Tumoral alfa/sangre , Adulto JovenRESUMEN
Hericium erinaceus, an edible and medicinal mushroom, displays various pharmacological activities in the prevention of dementia in conditions such as Parkinson's and Alzheimer's disease. The present study explored the neuroprotective effects of H. erinaceus mycelium polysaccharide-enriched aqueous extract (HE) on an l-glutamic acid (l-Glu)-induced differentiated PC12 (DPC12) cellular apoptosis model and an AlCl3 combined with d-galactose-induced Alzheimer's disease mouse model. The data revealed that HE successfully induced PC12 cell differentiation. A 3 h HE incubation at doses of 50 and 100 µg/mL before 25 mM of l-Glu effectively reversed the reduction of cell viability and the enhancement of the nuclear apoptosis rate in DPC12 cells. Compared with l-Glu-damaged cells, in PC12 cells, HE suppressed intracellular reactive oxygen species accumulation, blocked Ca2+ overload and prevented mitochondrial membrane potential (MMP) depolarization. In the Alzheimer's disease mouse model, HE administration enhanced the horizontal and vertical movements in the autonomic activity test, improved the endurance time in the rotarod test, and decreased the escape latency time in the water maze test. It also improved the central cholinergic system function in the Alzheimer's mice, demonstrated by the fact that it dose-dependently enhanced the acetylcholine (Ach) and choline acetyltransferase (ChAT) concentrations in both the serum and the hypothalamus. Our findings provide experimental evidence that HE may provide neuroprotective candidates for treating or preventing neurodegenerative diseases.
Asunto(s)
Enfermedad de Alzheimer/fisiopatología , Basidiomycota/química , Diferenciación Celular/efectos de los fármacos , Ácido Glutámico/farmacología , Fármacos Neuroprotectores/farmacología , Acetilcolina/sangre , Acetilcolina/metabolismo , Cloruro de Aluminio , Compuestos de Aluminio , Enfermedad de Alzheimer/sangre , Enfermedad de Alzheimer/inducido químicamente , Animales , Apoptosis/efectos de los fármacos , Western Blotting , Calcio/metabolismo , Supervivencia Celular/efectos de los fármacos , Cloruros , Colina O-Acetiltransferasa/sangre , Colina O-Acetiltransferasa/metabolismo , Femenino , Galactosa , Hipotálamo/efectos de los fármacos , Hipotálamo/metabolismo , Aprendizaje por Laberinto/efectos de los fármacos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Ratones Endogámicos BALB C , Actividad Motora/efectos de los fármacos , Micelio/química , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Fármacos Neuroprotectores/aislamiento & purificación , Células PC12 , Ratas , Especies Reactivas de Oxígeno/metabolismoRESUMEN
INTRODUCTION: There are a lot of pathogenic factors involved in development of polyetiologic diseases. Acetylcholine is known as first-order mediator and plays important role in development and maintenance of pathological processes. In this article we provide data on concentration of acetylcholine (Ah) in blood serum of patients with stomach ulcer (SU), duodenal ulcer (DU), alcoholic chronic pancreatitis (ACP) and control group.as well as activity of cholinesterase (Che). The aim of this study was to identify a role played by Ah in pathological process during a disease, that may complicate a course of the disease as poor prognostic factor. RESULTS AND DISCUSSION: We found that there are significant differences in Ah concentration and Che activity between SU, DU, ACP and control. In control group we divide concentration of Ah into three groups: low - 0.46 to 1.0 mlMol/l (60% of individuals), moderate - mlMol 02-1,5/I (30%) and high - more than 1.5 Ah mlMol/I (10%). CONCLUSION: We suppose that Che activity and Ah concentration depend on localization of pathological process. It is possible that there are hidden differences in Ah concentration and Che activity between GU and DU.
Asunto(s)
Acetilcolina/sangre , Acetilcolinesterasa/sangre , Úlcera Duodenal/sangre , Pancreatitis Alcohólica/sangre , Úlcera Gástrica/sangre , Adulto , Enfermedad Crónica , Femenino , Proteínas Ligadas a GPI/sangre , Humanos , MasculinoRESUMEN
Determining the concentrations of acetylcholine (ACh) and choline (Ch) is clinically important. ACh is a neurotransmitter that acts as a key link in the communication between neurons in the spinal cord and in nerve skeletal junctions in vertebrates, and plays an important role in transmitting signals in the brain. A bienzymatic sensor for the detection of ACh was prepared by co-immobilizing choline oxidase (ChO) and acetylcholinesterase (AChE) on graphene matrix/platinum nanoparticles, and then electrodepositing them on an ITO-coated glass plate. Graphene nanoparticles were decorated with platinum nanoparticles and were electrodeposited on a modified ITO-coated glass plate to form a modified electrode. The modified electrode was characterized by scanning electron microscopy (SEM), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) studies. The optimum response of the enzyme electrode was obtained at pH 7.0 and 35 °C. The response time of this ACh-sensing system was shown to be 4 s. The linear range of responses to ACh was 0.005-700 µM. This biosensor exhibits excellent anti-interferential abilities and good stability, retaining 50% of its original current even after 4 months. It has been applied for the detection of ACh levels in human serum samples.
Asunto(s)
Acetilcolina/sangre , Técnicas Biosensibles/métodos , Grafito/química , Nanopartículas/química , Platino (Metal)/química , Acetilcolina/metabolismo , Acetilcolinesterasa/metabolismo , Alcaligenes/enzimología , Oxidorreductasas de Alcohol/metabolismo , Enfermedad de Alzheimer/sangre , Animales , Espectroscopía Dieléctrica/métodos , Técnicas Electroquímicas/métodos , Electrodos , Electrophorus , Enzimas Inmovilizadas/metabolismo , Humanos , Límite de Detección , Modelos MolecularesRESUMEN
BACKGROUND: Activation of efferent vagus fibers exerts an anti-inflammatory role, a mechanism known as the cholinergic anti-inï¬ammatory pathway. We hypothesised that stimulation of atrial ganglionated plexi (GP) may also be anti-inflammatory. METHODS: Six-hour low-intensity GP stimulation was performed in eight dogs and the serum levels of acetylcholine (Ach), C reactive protein (CRP), interleukin-6 (IL-6), high-mobility group box 1 (HMGB1) were determined with enzyme-linked immunosorbent assay kits. RESULTS: The serum level of acetylcholine was significantly increased (P<0.05) while the serum levels of inflammatory factors of C reactive protein, interleukin-6 and high-mobility group box 1 were markedly decreased after six-hour GP stimulation (all P<0.05). CONCLUSIONS: These results suggest that GP stimulation exerts an anti-inflammatory role and might be a therapeutic option for inflammatory heart diseases.
Asunto(s)
Acetilcolina/sangre , Terapia por Estimulación Eléctrica , Proteína HMGB1/sangre , Sistema de Conducción Cardíaco , Mediadores de Inflamación/sangre , Interleucina-6/sangre , Miocarditis , Animales , Perros , Femenino , Masculino , Miocarditis/sangre , Miocarditis/terapiaRESUMEN
We studied the responses of the organism to administration of autonomic transmitters in different doses: epinephrine (from 0.0000001 to 300 µg per 100 g body weight), serotonin (25, 50, 250, and 500 µg), and their combinations. The concentrations of biogenic amines in the blood and hypothalamus and parameters of oxidative and antioxidant systems, immune status, and blood coagulation and anticoagulation systems were analyzed. Cyclic dose-dependent variations in the levels of transmitters were revealed that dynamically changed their properties of syntoxins and catatoxins.