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1.
PLoS One ; 18(4): e0283995, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37027451

RESUMEN

Xerostomia is a subjective condition of dryness of the oral cavity that may lead to several oral problems deteriorating oral health-related quality of life. This study aimed to (1) determine the prevalence of xerostomia, (2) compare the general health status, unstimulated salivary flow rate, and oral health-related quality of life in xerostomics and non-xerostomics, and (3) investigate the potential of salivary aquaporin-3 (AQP-3) as a screening biomarker for xerostomia in patients with periodontal disease. Demographics and systemic health data were collected from 109 healthy participants, 20 to 55 years old, with Community Periodontal Index (CPI) score ≥ 3. For subjective assessment of xerostomia, Shortened Xerostomia Inventory (SXI) was used. For objective assessment of xerostomia, unstimulated salivary flow rate was measured. Shortened Oral Health Impact Profile (S-OHIP) was utilized for oral health-related quality of life assessment. The collected saliva samples were processed and stored at -80°C. Quantification of salivary AQP-3 protein was done with enzyme-linked immunosorbent assay. Xerostomia was reported in 78% of the subjects based on SXI score. Median concentration of AQP-3 was significantly higher in xerostomics compared to non-xerostomics, p = 0.001. Moreover, oral health-related quality of life was significantly poor in xerostomics compared to non-xerostomics, p = 0.002. Furthermore, there were significant correlations between AQP-3 and SXI (r = 0.21, p = 0.025), AQP-3 and S-OHIP (r = 0.2, p = 0.042), S-OHIP and SXI (r = 0.37, p < 0.001), unstimulated salivary flow rate and random blood glucose level (r = 0.32, p = 0.001), and body mass index and mean arterial pressure (r = 0.44, p < 0.001). Regression analysis showed that body mass index, CPI score 3, and salivary AQP-3 were suitable predictors for presence of xerostomia. AQP-3 could be a potential screening biomarker for xerostomia in patients with periodontal disease for its early identification may help improve oral health-related quality of life of the individuals.


Asunto(s)
Acuaporina 3 , Acuaporinas , Enfermedades Periodontales , Xerostomía , Adulto , Humanos , Persona de Mediana Edad , Adulto Joven , Acuaporinas/metabolismo , Enfermedades Periodontales/epidemiología , Calidad de Vida , Saliva/química , Xerostomía/diagnóstico , Xerostomía/epidemiología , Acuaporina 3/análisis , Salud Bucal
2.
Biol Reprod ; 106(1): 173-184, 2022 01 13.
Artículo en Inglés | MEDLINE | ID: mdl-34664639

RESUMEN

Despite aquaporin water channels (AQPs) play a critical role in maintaining water homeostasis in female reproductive tract and prompt a gradual increase in water content in cervical edema as pregnancy progressed, their relationship with macrophage infiltration and collagen content in human cervical remodeling need to be further investigated. This is the first study to examine the expression and localization of AQP3, AQP4, AQP5, AQP8, and macrophages simultaneously in human cervical ripening. The immunoreactivity of these AQPs was 2.6 to 6-fold higher on gestational weeks 26 (GD26W) than that on GD6W and GD15W, but AQP4 expression on GD39W dropped a similar extent on GD15W, other AQPs continued to rise on GD39W. The AQP3, AQP4, and AQP5 intensity seemed more abundant in cervical stroma than in the perivascular area on GD26W; the distribution of AQP3, AQP5, and AQP8 in cervical stroma was equivalent to that in the perivascular area on GD39W. Macrophage numbers were 1.7-fold higher in subepithelium region and 3.0-fold higher in center area on GD26W than that on GD15W; such numbers remained elevated on GD39W. The electron micrographs showed that cervical extensibility increased significantly on GD26W and GD39W accompanied with increased macrophage infiltration, cervical water content, and much more space among collagen fibers. These findings suggest that the upregulation of AQPs expression in human cervix is closely related to enhanced macrophage infiltration during pregnancy; there may be a positive feedback mechanism between them to lead the increase of water content and the degradation of collagen.


Asunto(s)
Acuaporinas/análisis , Cuello del Útero/fisiología , Macrófagos/fisiología , Adolescente , Adulto , Acuaporina 3/análisis , Acuaporina 4/análisis , Acuaporina 5/análisis , Acuaporinas/fisiología , Recuento de Células , Maduración Cervical/fisiología , Cuello del Útero/química , Cuello del Útero/citología , Colágeno/análisis , Colágeno/metabolismo , Femenino , Edad Gestacional , Humanos , Macrófagos/ultraestructura , Microscopía Electrónica , Embarazo , Adulto Joven
3.
Anal Cell Pathol (Amst) ; 2021: 7866471, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34745849

RESUMEN

Aquaporin 3 (AQP3) is the membrane channel of water and involved in fluid homeostasis. The aim of this study was to reveal the expression and significance of AQP3 in cutaneous lesions. We analyzed AQP3 mRNA levels using RT-PCR in 311 cutaneous lesions and confirmed AQP3 expression in these lesions by immunohistochemistry. AQP3 mRNA was detected in normal epidermis, seborrheic keratosis, solar keratosis, Bowen's disease, squamous cell carcinoma, eccrine poroma, apocrine carcinoma, and sebaceoma; however, AQP3 mRNA was absent in basal cell carcinoma, nevocellular nevus, or malignant melanoma. By immunohistochemistry, diffuse AQP3 expression was seen in all keratotic lesions including seborrheic keratosis, verruca vulgaris, molluscum contagiosum, solar keratosis, Bowen's disease, and squamous cell carcinoma. Diffuse AQP3 expression was also present in all extramammary Paget's disease. No AQP3 staining was obtained in basal cell carcinoma. Positive AQP3 staining was seen in sweat gland tumors including hidradenoma, eccrine poroma, and apocrine carcinoma. Among sebaceous tumors, AQP3 expressed diffusely in all sebaceous hyperplasia and sebaceous adenoma, but not in sebaceous carcinomas. Only focal AQP3 staining was seen in nevocellular nevus and no AQP3 staining in melanoma. Our findings indicate the function of AQP3 maintained in most skin tumors. AQP3 may be used for differential diagnosis in skin tumors.


Asunto(s)
Acuaporina 3/biosíntesis , Biomarcadores de Tumor/análisis , Biomarcadores de Tumor/metabolismo , Neoplasias Cutáneas/diagnóstico , Acuaporina 3/análisis , Diagnóstico Diferencial , Humanos , Enfermedades de la Piel/diagnóstico , Enfermedades de la Piel/metabolismo , Neoplasias Cutáneas/metabolismo
4.
Int J Legal Med ; 135(3): 903-908, 2021 May.
Artículo en Inglés | MEDLINE | ID: mdl-33447890

RESUMEN

In sauna-associated deaths, the vitality of heat exposure is of great importance. Two case reports address this. First, we present the case of a 77-year-old man who was found dead in the sauna of his family home. When found, the sauna door was closed, and the sauna indicated a temperature of 78 °C. The body had already begun to decay and was partially mummified when it was found. In the other case, a 73-year-old woman was found dead in the sauna by her husband. In this case, the sauna door was also closed. The sauna was still in operation at a temperature of approximately 70 °C. Epidermal detachments were found. In both autopsies and their follow-up examinations, there were no indications of a cause of death competing with heat shock. The expression of heat shock proteins in kidneys and lungs and the expression of aquaporin 3 in skin were investigated to detect pre-mortal temperature influences.


Asunto(s)
Acuaporina 3/análisis , Proteínas de Choque Térmico/análisis , Respuesta al Choque Térmico , Riñón/patología , Pulmón/patología , Piel/patología , Baño de Vapor , Anciano , Resultado Fatal , Femenino , Humanos , Masculino
5.
Zhongguo Zhong Yao Za Zhi ; 45(3): 623-630, 2020 Feb.
Artículo en Chino | MEDLINE | ID: mdl-32237522

RESUMEN

The aim of this paper was to investigate the effect of Faeces Bombycis(FB) on the intestinal microflora in rats with syndrome of damp retention in middle-jiao, and to explore its mechanism in regulating intestinal microflora from the perspective of microorganisms contained in FB. The contents of antidiuretic hormone(ADH) and C-reactive protein(CRP) in serum and aquaporin 3(AQP3) in jejunum were determined by enzyme-linked immunosorbent assay(ELISA). Illumina Miseq platform was used for high-throughput sequencing of the rat feces and FB. The ELISA results showed that as compared with the normal control group, the contents of ADH and CRP in the model group were significantly increased(P<0.05), and the content of AQP3 was significantly decreased(P<0.05). After drug administration, the ADH, CRP and AQP3 contents were recovered. Sequencing of rat feces showed that the ACE, Chao1 and Shannon indexes of the intestinal microflora were the lowest in the model group. As compared with the normal control group, the levels from phylum to genus were all significantly changed in model group, and Proteobacteria, Acinetobacter, Anaerobacter, Pseudomonas, and Parabacteroides levels were significantly increased(P<0.05), while Marvinbryantia level was significantly decreased(P<0.05). As compared with the model group, Proteobacteria was significantly decreased in the FB low and high dose groups(P<0.05), and Acinetobacter, Anaerobacter, Pseudomonas, Parabacteroides levels were significantly decreased in the low, medium and high dose groups(P<0.05), while Lachnoanaerobaculum, Intestinimonas and Marvinbryantia were increased significantly in the high dose group(P<0.05). Sequencing analysis of FB showed that the relative abundance of Leclercia, Pantoea, Brachybacterium, Shimwellia, Hartmannibacter, Klebsiella, Serratia, Aurantimonas, Paenibacillus and Bacillus was high in the FB, but they were basically not present or little in the rat feces. In conclusion, FB may play a role in the treatment of "syndrome of damp retention in middle-jiao" by balancing the intestinal microflora, and this effect may be related to the metabolites of microorganisms in the FB.


Asunto(s)
Bombyx/química , Heces/química , Microbioma Gastrointestinal , Animales , Acuaporina 3/análisis , Proteína C-Reactiva/análisis , Secuenciación de Nucleótidos de Alto Rendimiento , Medicina Tradicional China , Ratas , Vasopresinas/sangre
6.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-1008547

RESUMEN

The aim of this paper was to investigate the effect of Faeces Bombycis(FB) on the intestinal microflora in rats with syndrome of damp retention in middle-jiao, and to explore its mechanism in regulating intestinal microflora from the perspective of microorganisms contained in FB. The contents of antidiuretic hormone(ADH) and C-reactive protein(CRP) in serum and aquaporin 3(AQP3) in jejunum were determined by enzyme-linked immunosorbent assay(ELISA). Illumina Miseq platform was used for high-throughput sequencing of the rat feces and FB. The ELISA results showed that as compared with the normal control group, the contents of ADH and CRP in the model group were significantly increased(P<0.05), and the content of AQP3 was significantly decreased(P<0.05). After drug administration, the ADH, CRP and AQP3 contents were recovered. Sequencing of rat feces showed that the ACE, Chao1 and Shannon indexes of the intestinal microflora were the lowest in the model group. As compared with the normal control group, the levels from phylum to genus were all significantly changed in model group, and Proteobacteria, Acinetobacter, Anaerobacter, Pseudomonas, and Parabacteroides levels were significantly increased(P<0.05), while Marvinbryantia level was significantly decreased(P<0.05). As compared with the model group, Proteobacteria was significantly decreased in the FB low and high dose groups(P<0.05), and Acinetobacter, Anaerobacter, Pseudomonas, Parabacteroides levels were significantly decreased in the low, medium and high dose groups(P<0.05), while Lachnoanaerobaculum, Intestinimonas and Marvinbryantia were increased significantly in the high dose group(P<0.05). Sequencing analysis of FB showed that the relative abundance of Leclercia, Pantoea, Brachybacterium, Shimwellia, Hartmannibacter, Klebsiella, Serratia, Aurantimonas, Paenibacillus and Bacillus was high in the FB, but they were basically not present or little in the rat feces. In conclusion, FB may play a role in the treatment of "syndrome of damp retention in middle-jiao" by balancing the intestinal microflora, and this effect may be related to the metabolites of microorganisms in the FB.


Asunto(s)
Animales , Ratas , Acuaporina 3/análisis , Bombyx/química , Proteína C-Reactiva/análisis , Heces/química , Microbioma Gastrointestinal , Secuenciación de Nucleótidos de Alto Rendimiento , Medicina Tradicional China , Vasopresinas/sangre
7.
Sci Rep ; 9(1): 16639, 2019 11 12.
Artículo en Inglés | MEDLINE | ID: mdl-31719660

RESUMEN

Lepus yarkandensis is a desert hare of the Tarim Basin in western China, and it has strong adaptability to arid environments. Aquaporins (AQPs) are a family of water channel proteins that facilitate transmembrane water transport. Gastrointestinal tract AQPs are involved in fluid absorption in the small intestine and colon. This study aimed to determine the distribution of AQPs and sodium transporters in the gastrointestinal tract of L. yarkandensis and to compare the expression of these proteins with that in Oryctolagus cuniculus. Immunohistochemistry was performed to analyse the cellular distribution of these proteins, and the acquired images were analysed with IpWin32 software. Our results revealed that AQP1 was located in the colonic epithelium, central lacteal cells, fundic gland parietal cells, and capillary endothelial cells; AQP3 was located in the colonic epithelium, small intestinal villus epithelium, gastric pit and fundic gland; AQP4 was located in the fundic gland, small intestinal gland and colonic epithelium; and epithelial sodium channel (ENaC) and Na+-K+-ATPase were located in the epithelial cells, respectively. The higher expression levels of AQP1, AQP3, ENaC and Na+-K+-ATPase in the colon of L. yarkandensis compared to those in O. cuniculus suggested that L. yarkandensis has a higher capacity for faecal dehydration.


Asunto(s)
Acuaporinas/análisis , Canales Epiteliales de Sodio/análisis , Tracto Gastrointestinal/química , ATPasa Intercambiadora de Sodio-Potasio/análisis , Animales , Acuaporina 1/análisis , Acuaporina 3/análisis , Colon/química , Clima Desértico , Liebres , Mucosa Intestinal/química , Intestino Grueso/química , Intestino Delgado/química , Masculino , Estómago/química
8.
Int J Mol Sci ; 20(15)2019 Aug 02.
Artículo en Inglés | MEDLINE | ID: mdl-31382467

RESUMEN

Xeroderma is a frequent complication in diabetic patients. In this study, we investigated the mechanism underlying the onset of diabetic xeroderma, focusing on aquaporin-3 (AQP3), which plays an important role in water transport in the skin. Dermal water content in diabetic mice was significantly lower than that in control mice. The expression level of AQP3 in the skin was significantly lower in diabetic mice than in control mice. One week after streptozotocin (STZ) treatment, despite their increased blood glucose levels, mice showed no changes in the expression levels of AQP3, Bmal1, Clock, and D site-binding protein (Dbp) in the skin and 8-hydroxydeoxyguanosine (8-OHdG) in the urine. In contrast, two weeks after STZ treatment, mice showed increases in the blood glucose level, decreases in AQP3, Bmal1, Clock, and Dbp levels, and increases in the urinary levels of 8-OHdG. The results of this study suggest that skin AQP3 expression decreases in diabetes, which may limit water transport from the vessel side to the corneum side, causing dry skin. In addition, in diabetic mice, increased oxidative stress triggered decreases in the expression levels of Bmal1 and Clock in the skin, thereby inhibiting the transcription of Aqp3 by Dbp, which resulted in decreased AQP3 expression.


Asunto(s)
Acuaporina 3/metabolismo , Diabetes Mellitus Experimental/complicaciones , Ictiosis/etiología , Agua/metabolismo , Animales , Acuaporina 3/análisis , Glucemia/análisis , Glucemia/metabolismo , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/patología , Modelos Animales de Enfermedad , Ingestión de Líquidos , Ictiosis/metabolismo , Ictiosis/patología , Masculino , Ratones , Ratones Pelados , Piel/metabolismo , Piel/patología , Estreptozocina , Agua/análisis
9.
Nano Lett ; 19(2): 699-707, 2019 02 13.
Artículo en Inglés | MEDLINE | ID: mdl-30584808

RESUMEN

Several aquaporin (AQP) water channels are short-term regulated by the messenger cyclic adenosine monophosphate (cAMP), including AQP3. Bulk measurements show that cAMP can change diffusive properties of AQP3; however, it remains unknown how elevated cAMP affects AQP3 organization at the nanoscale. Here we analyzed AQP3 nano-organization following cAMP stimulation using photoactivated localization microscopy (PALM) of fixed cells combined with pair correlation analysis. Moreover, in live cells, we combined PALM acquisitions of single fluorophores with single-particle tracking (spt-PALM). These analyses revealed that AQP3 tends to cluster and that the diffusive mobility is confined to nanodomains with radii of ∼150 nm. This domain size increases by ∼30% upon elevation of cAMP, which, however, is not accompanied by a significant increase in the confined diffusion coefficient. This regulation of AQP3 organization at the nanoscale may be important for understanding the mechanisms of water AQP3-mediated water transport across plasma membranes.


Asunto(s)
Acuaporina 3/metabolismo , Membrana Celular/metabolismo , AMP Cíclico/metabolismo , Células Epiteliales/metabolismo , Animales , Acuaporina 3/análisis , Membrana Celular/ultraestructura , Difusión , Perros , Células Epiteliales/ultraestructura , Células de Riñón Canino Madin Darby , Microscopía Fluorescente/métodos , Procesos Fotoquímicos
10.
Biol Cell ; 108(10): 294-305, 2016 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-27406921

RESUMEN

BACKGROUND INFORMATION: Anopheles gambiae is the major mosquito vector for Plasmodium falciparum malaria in sub-Saharan Africa, where it survives in stressful climates. Aquaporin water channels are expressed in all life forms, where they provide environmental adaptation by conferring rapid trans-cellular movement of water (classical aquaporins) or water plus glycerol (aquaglyceroporins). Here, we report an aquaglyceroporin homolog in A. gambiae, AgAQP3 (A. gambiae aquaglyceroporin 3). RESULTS: Despite atypical pore-lining amino acids, AgAQP3 is permeated by water, glycerol and urea, and is not significantly inhibited by 1 mM HgCl2 . AgAQP3 is expressed more heavily in male mosquitoes, yet adult female A. gambiae abundantly express AgAQP3 in Malpighian tubules and gut where large amounts of fluid exchange occur during blood meal digestion, water and nutrient absorption and waste secretion. Reducing expression of AgAQP3 by RNA interference reduces median mosquito survival at 39°C. After an infectious blood meal, mosquitoes with depleted AgAQP3 expression exhibit fewer P. falciparum oocysts in the midgut compared to control mosquitoes. CONCLUSIONS: Our studies reveal critical contributions of AgAQP3 to A. gambiae heat tolerance and P. falciparum development in vivo. SIGNIFICANCE: This study indicates that AgAQP3 may be a major factor explaining why A. gambiae is an important malaria vector mosquito in sub-Saharan Africa, and may be a potential target for novel malaria control strategies.


Asunto(s)
Anopheles/fisiología , Acuaporina 3/metabolismo , Proteínas de Insectos/metabolismo , Insectos Vectores/fisiología , Malaria Falciparum/transmisión , Plasmodium falciparum/parasitología , Adaptación Fisiológica , Animales , Anopheles/genética , Acuaporina 3/análisis , Acuaporina 3/genética , Femenino , Calor , Humanos , Proteínas de Insectos/análisis , Insectos Vectores/genética , Masculino , Interferencia de ARN , ARN Interferente Pequeño/genética , Estrés Fisiológico
11.
Ultrastruct Pathol ; 39(5): 306-17, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26107428

RESUMEN

Aquaporin-3 (AQP3), is an aquaglyceroporin, that plays a role in cell proliferation, tumorigenesis, and cell migration. This study aimed at evaluating the possible role of AQP3 in nonmelanoma skin cancer (NMSC) pathogenesis through its immunohistochemical expression in skin biopsies of these diseases. One-hundred and thirty cutaneous specimens were studied. These included 60 cases of NMSC and 40 normal skin and 30 psoriasis samples, from age- and gender-matched subjects, as a control group. AQP3 was expressed in 66.7% of basal cell carcinoma (BCC) cases and in 93.3% of squamous cell carcinoma (SCC) cases. Higher AQP3 expression (p = .01), expression percentage (p = .01), and H score (p = .04) were significantly associated with SCC compared to BCC. Normal skin and psoriasis showed significantly higher AQP3 expression (p = .001, p < .001, respectively), expression percentage (p < .001 for both), and H score values (p < .001, p = .001, respectively) compared to NMSC. Higher H score values in BCC were significantly associated with female gender (p = .02) and with nodular lesions (p > .001). Higher H score values in SCC were significantly associated with grade III tumors (p = .04) and AQP3 percentage of expression was significantly correlated with grade III tumors (r = .48, p = .009). In conclusion, AQP3 may play a role in NMSC pathogenesis. This probably occurs through aquaporin-mediated glycerol transport and ATP generation. Its downregulation, observed in the current work, is mostly a result of excessive proliferation. Further studies are needed to investigate the therapeutic effect of its inhibition in NMSC treatment.


Asunto(s)
Acuaporina 3/biosíntesis , Carcinoma de Células Escamosas/metabolismo , Neoplasias Basocelulares/metabolismo , Neoplasias Cutáneas/metabolismo , Adulto , Anciano , Acuaporina 3/análisis , Carcinoma de Células Escamosas/patología , Estudios de Casos y Controles , Femenino , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Neoplasias Basocelulares/patología , Neoplasias Cutáneas/patología
12.
Theriogenology ; 84(1): 1-10, 2015 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-25930733

RESUMEN

Aquaporins (AQPs) are a well-conserved family of small (approximately 30 kDa) membrane channel proteins that facilitate rapid movement of fluids and have a unique tissue-specific pattern of expression. These proteins have been found in the female reproductive systems of humans, rats, and mice. However, the expression and cellular localization of AQPs have not extensively been studied in the female reproductive system of sheep. Therefore, this study aimed to evaluate, by real-time polymerase chain reaction and immunohistochemistry respectively, the levels of messenger RNA and the immunolocalization of AQP3, AQP7, and AQP9 in large isolated ovine secondary follicles over a period of IVC. Our analysis revealed that AQP3 and AQP9 were present predominately in follicles that exhibited antrum formation, suggesting a crucial role of these AQPs in the formation of the antrum. Interestingly, AQP7 was only expressed in follicles that had not formed an antrum by Day 12 of culture. In conclusion, the presence of protein channels (AQP3 and AQP9) seems to be essential for the formation of the antrum in isolated ovine secondary follicles cultured in vitro and thus plays an important role during folliculogenesis in this species.


Asunto(s)
Acuaporina 3/metabolismo , Acuaporinas/metabolismo , Folículo Ovárico/metabolismo , ARN Mensajero/metabolismo , Ovinos/metabolismo , Animales , Acuaporina 3/análisis , Acuaporinas/análisis , Técnicas de Cultivo de Célula , Femenino , Inmunohistoquímica , Folículo Ovárico/citología , Folículo Ovárico/crecimiento & desarrollo
13.
Biomed Res Int ; 2014: 619465, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-25215286

RESUMEN

Tannins, a group of major active components of Chinese rhubarb and widely distributed in nature, have a significant antidiarrhoeal activity. Aquaporins (AQPs) 2 and 3 play important roles in regulating water transfer during diarrhoea. The present study aims to determine the effect of the total tannins extract of rhubarb on aquaporins (AQPs) 2 and 3 in diarrhoea mice and HT-29 cells both induced by magnesium sulphate (MgSO4). Our results showed that rhubarb tannins extract (RTE) significantly decreased the faecal water content in colon and evaluation index of defecation of diarrhoea mice. Interestingly, RTE could markedly reduce the mRNA and protein expression levels of AQPs 2 and 3 in apical and lateral mucosal epithelial cells in the colons of diarrhoea mice and HT-29 cells both induced by MgSO4 in a dose-dependent manner. Furthermore, RTE suppressed the production of cyclic monophosphate- (cAMP-) dependent protein kinase A catalytic subunits α (PKA C-α) and phosphorylated cAMP response element-binding protein (p-CREB, Ser133) in MgSO4-induced HT-29 cells. Our data showed for the first time that RTE inhibit AQPs 2 and 3 expression in vivo and in vitro via downregulating PKA/p-CREB signal pathway, which accounts for the antidiarrhoeal effect of RTE.


Asunto(s)
Acuaporina 2/metabolismo , Acuaporina 3/metabolismo , Diarrea/metabolismo , Extractos Vegetales/farmacología , Rheum/química , Taninos/farmacología , Animales , Acuaporina 2/análisis , Acuaporina 2/genética , Acuaporina 3/análisis , Acuaporina 3/genética , Supervivencia Celular/efectos de los fármacos , Diarrea/inducido químicamente , Modelos Animales de Enfermedad , Expresión Génica/efectos de los fármacos , Células HT29 , Humanos , Sulfato de Magnesio/efectos adversos , Ratones , Ratones Endogámicos ICR , Extractos Vegetales/química , Taninos/química
14.
J Vis Exp ; (87)2014 May 10.
Artículo en Inglés | MEDLINE | ID: mdl-24893770

RESUMEN

Lateral diffusion and compartmentalization of plasma membrane proteins are tightly regulated in cells and thus, studying these processes will reveal new insights to plasma membrane protein function and regulation. Recently, k-Space Image Correlation Spectroscopy (kICS)(1) was developed to enable routine measurements of diffusion coefficients directly from images of fluorescently tagged plasma membrane proteins, that avoided systematic biases introduced by probe photophysics. Although the theoretical basis for the analysis is complex, the method can be implemented by nonexperts using a freely available code to measure diffusion coefficients of proteins. kICS calculates a time correlation function from a fluorescence microscopy image stack after Fourier transformation of each image to reciprocal (k-) space. Subsequently, circular averaging, natural logarithm transform and linear fits to the correlation function yields the diffusion coefficient. This paper provides a step-by-step guide to the image analysis and measurement of diffusion coefficients via kICS. First, a high frame rate image sequence of a fluorescently labeled plasma membrane protein is acquired using a fluorescence microscope. Then, a region of interest (ROI) avoiding intracellular organelles, moving vesicles or protruding membrane regions is selected. The ROI stack is imported into a freely available code and several defined parameters (see Method section) are set for kICS analysis. The program then generates a "slope of slopes" plot from the k-space time correlation functions, and the diffusion coefficient is calculated from the slope of the plot. Below is a step-by-step kICS procedure to measure the diffusion coefficient of a membrane protein using the renal water channel aquaporin-3 tagged with EGFP as a canonical example.


Asunto(s)
Proteínas Fluorescentes Verdes/química , Proteínas de la Membrana/química , Microscopía Fluorescente/métodos , Proteínas Recombinantes de Fusión/química , Animales , Acuaporina 3/análisis , Acuaporina 3/química , Perros , Proteínas Fluorescentes Verdes/análisis , Procesamiento de Imagen Asistido por Computador , Células de Riñón Canino Madin Darby , Proteínas Recombinantes de Fusión/análisis
15.
Bull Tokyo Dent Coll ; 55(1): 1-10, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24717924

RESUMEN

Aquaporins (AQPs) are a family of small integral membrane proteins made up of 6 hydrophobic, a-helical, membrane-spanning domains surrounding a highly selective aqueous pore. AQP3, AQP7, and AQP9, termed aqua-glyceroporins, are known to be involved in the transport of water, glycerol, and other small molecules. In this study, we investigated the expression and localization of aqua-glyceroporins in rat oral stratified squamous epithelia of the palate, the buccal mucosa, the inferior aspect of the tongue, and the oral floor by using RT-PCR, immunofluorescence, and immunogold electron microscopy. AQP3 and AQP9 mRNAs were expressed in whole oral epithelium. Immunostaining for AQP3 was recognized in each type of epithelium. The results suggest that AQP3 synthesis begins predominantly in the cytoplasm of the basal cells. During the process of epithelial cell differentiation, AQP3 protein appears to accumulate and be transported to the plasma membrane, from where it is incorporated into the cornified or surface layers. The intracellular localization of AQP3 appears to correlate with the differentiation of keratinocytes, suggesting that it acts as an enhancer of the physiological permeability barrier together with membrane coating granules. The distribution pattern of AQP9 was limited to the marginal areas of the basal and suprabasal layers, which was different from that of AQP3. This difference in distribution between AQP3 and AQP9 suggests that AQP9 in rat oral epithelia acts as a channel by facilitating glycerol uptake from the blood through the endothelial cells of the capillary vessels to the oral stratified squamous epithelium. AQP3 and AQP9 facilitate both transcellular osmotic water flow and glycerol transport as pore-like passive transporters in the keratinocytes of oral epithelia, and may play a key role in not only hydration and the permeability barrier, but also cell proliferation, differentiation, migration, development, and wound healing by generating ATP.


Asunto(s)
Acuaporina 3/análisis , Acuaporinas/análisis , Mucosa Bucal/química , Animales , Diferenciación Celular/fisiología , Membrana Celular/química , Permeabilidad de la Membrana Celular/fisiología , Mejilla , Citoplasma/química , Células Endoteliales/metabolismo , Células Epiteliales/química , Epitelio/química , Glicerol/sangre , Glicerol/metabolismo , Queratinocitos/química , Masculino , Suelo de la Boca/química , Ósmosis/fisiología , Hueso Paladar/química , Ratas , Lengua/química
16.
Acta Odontol Scand ; 72(7): 549-56, 2014 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-24471729

RESUMEN

OBJECTIVE: Sjögren's Syndrome (SS) is a chronic autoimmune disease, leading to deficient secretion from salivary and lacrimal glands. Saliva production is normally increased by cholinergic innervation, giving rise to intracellular calcium signaling and water transport through water channels (aquaporins, AQPs). The aim of this study was to investigate possible pathophysiological changes in cell volume regulation, AQP expression and localization, and intracellular calcium signaling in glandular cells from SS patients compared to controls. MATERIALS AND METHODS: A total of 35 SS patients and 41 non-SS controls were included. Real time qPCR was combined with immunohistochemistry to analyze the mRNA expression and cellular distribution of AQP1, 3 and 5. Cell volume regulation and intracellular calcium signaling were examined in fresh acinar cells. RESULTS: We show for the first time a reduced mRNA expression of AQP1 and 5 in SS compared to controls, accompanied by a decrease in staining intensity of AQP1, 3 and 5 in areas adjacent to local lymphocytic infiltration. Furthermore, we observed that the SS cells' capacity for volume regulation was abnormal. Similarly, the calcium response after parasympathetic agonist (carbachol) stimulation was markedly decreased in SS cells. CONCLUSIONS: It is concluded that mRNA expression of AQP1 and 5, protein distribution of AQP1, 3 and 5, glandular cell volume regulation and intracellular calcium signaling are all altered in SS, pointing to possible pathophysiological mechanisms in SS.


Asunto(s)
Señalización del Calcio/fisiología , Glándulas Salivales/patología , Síndrome de Sjögren/fisiopatología , Adulto , Anciano , Anciano de 80 o más Años , Acuaporina 1/análisis , Acuaporina 3/análisis , Acuaporina 5/análisis , Acuaporinas/análisis , Señalización del Calcio/efectos de los fármacos , Carbacol/farmacología , Tamaño de la Célula , Agonistas Colinérgicos/farmacología , Estudios de Cohortes , Femenino , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Saliva/química , Saliva/metabolismo , Glándulas Salivales/efectos de los fármacos , Glándulas Salivales/metabolismo , Método Simple Ciego , Síndrome de Sjögren/patología
17.
Yakugaku Zasshi ; 133(9): 955-61, 2013.
Artículo en Japonés | MEDLINE | ID: mdl-23995803

RESUMEN

Aquaporins (AQPs) are membrane channels that transport water within the human body and are therefore important for the regulation of water homeostasis. However, little is known regarding the details of the physiological role of AQP3, which is predominantly expressed in the colon. Thus, we investigated the role of AQP3 in the colon using laxative agents (magnesium sulfate and bisacodyl). The results suggest that the laxative effect produced by magnesium sulfate, which is classified as an osmotic laxative, is not simply a result of the changes in osmotic pressure but is also associated with the increased expression of AQP3 in the mucosal epithelial cells of the colon. In addition, magnesium sulfate increased colonic AQP3 expression through adenylate cyclase activation, which is caused by an increase in the intracellular Mg(2+) concentration. This effect may trigger CREB phosphorylation through PKA activation and promote AQP3 gene transcription. Meanwhile, bisacodyl, which is classified as a stimulant laxative, decreases the expression level of AQP3 in the mucosal epithelial cells of the colon, resulting in the inhibition of water transfer from the intestinal tract to the vascular side of the epithelium, eventually leading to the development of diarrhea. It was also observed that the direct activation of colon macrophages by bisacodyl increases the secretion of PGE2, which acts as a paracrine factor and decreases AQP3 expression in colon mucosal epithelial cells. Future studies of the enteric AQP3 expression level and water transport may aid in the development of new laxative and antidiarrheal agents that target AQP3.


Asunto(s)
Acuaporina 3/fisiología , Colon/química , Animales , Acuaporina 3/análisis , Bisacodilo/administración & dosificación , Bisacodilo/farmacología , Humanos , Mucosa Intestinal/química , Laxativos/administración & dosificación , Laxativos/farmacología , Sulfato de Magnesio/administración & dosificación , Sulfato de Magnesio/farmacología , Ratas
18.
J Cosmet Dermatol ; 12(2): 86-95, 2013 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-23725301

RESUMEN

BACKGROUND: As oxygen is essential for wound healing and there is limited diffusion across the stratum corneum into the epidermis, we wanted to evaluate whether the topical delivery of a total dissolved oxygen in dressing form on intact human subject skin would improve clinical and histologic skin functioning. AIMS: Fifty normal, healthy subjects completed a pilot clinical evaluation to assess the efficacy and tolerability of a dissolved oxygen dressing (OxygeneSys™-Continuous) to improve the health and appearance of intact skin. METHODS: Clinical analysis was performed on 50 subjects; histological and gene expression analysis was performed on 12 of the 50 subjects to assess the effect of the dissolved oxygen dressing. RESULTS: Clinical data demonstrate that the dressing is well tolerated, and several measures of skin health and integrity showed improvements compared with a control dressing site. Skin hydration measurements showed a statistically significant increase in skin hydration at 0-4, 4-8, and 0-8 weeks (P < 0.05 at each time point). The blinded clinical investigator's grading of desquamation, roughness, and skin texture show significant decreases from baseline to the 8-week time point (P < 0.05). The dressings were removed prior to the blinded clinical investigator's grading. These data were supported by the histological and gene expression studies, which showed a general reduction in inflammatory response markers and transcription products (IL-6, IL-8, TNF-alpha, MMP-1, and MMP-12), while facilitating a general increase in structural skin proteins (collagen I, elastin, and filaggrin). Additionally, p53 signals from biopsy samples support the clinical investigator's observations of no safety concerns. CONCLUSION: The data from this study demonstrate that the dressing has no deleterious effects and stimulates beneficial effects on intact, nonwounded skin.


Asunto(s)
Expresión Génica/efectos de los fármacos , Oxígeno/farmacología , Fenómenos Fisiológicos de la Piel/efectos de los fármacos , Piel/efectos de los fármacos , 8-Hidroxi-2'-Desoxicoguanosina , Administración Cutánea , Anciano , Acuaporina 3/análisis , Colágeno Tipo I/genética , Desoxiguanosina/análogos & derivados , Desoxiguanosina/análisis , Elastina/genética , Femenino , Proteínas Filagrina , Humanos , Inflamación/genética , Interleucina-1/genética , Interleucina-6/genética , Proteínas de Filamentos Intermediarios/genética , Masculino , Metaloproteinasa 1 de la Matriz/genética , Metaloproteinasa 12 de la Matriz/genética , Persona de Mediana Edad , Oxígeno/administración & dosificación , Método Simple Ciego , Piel/anatomía & histología , Piel/química , Factor de Necrosis Tumoral alfa/genética
19.
Acta Histochem ; 115(3): 257-63, 2013 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-22889702

RESUMEN

Sea urchins are echinoderms, marine invertebrates found at the base of the deutorostome lineage, which show separate sexes and are external spawners. In the sea urchin, efficient regulation of water homeostasis is essential for many biological processes such as cellular respiration, normal fertilization and correct embryo growth. In order to clarify some of these processes, the present study reports on the identification and function of aquaporin proteins in the sea urchin. Our results show, by immunoblot, immunoelectron microscopy and immunofluorescence analysis, the presence of aquaporin1- and aquaporin3-like proteins in virgin eggs and in early embryogenesis of Paracentrotus lividus and, by using known inhibitors of aquaporin functions, the functional and relevant role of aquaporin-3 in the fertilization process. AQP3 in particular seems to play a crucial role in high velocity water flux formations involved in the detachment of the vitelline layer during the slow block of polyspermy, while the presence of AQP1 and the increase of AQP3 in the first phase of the P. lividus developmental cycle, suggest their involvement in the appropriate homeostasis for embryo development.


Asunto(s)
Acuaporina 1/análisis , Acuaporina 3/análisis , Oocitos/metabolismo , Paracentrotus/embriología , Paracentrotus/metabolismo , Animales , Acuaporina 1/metabolismo , Acuaporina 3/metabolismo , Oocitos/crecimiento & desarrollo
20.
J Sex Med ; 10(2): 342-9, 2013 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-23110393

RESUMEN

INTRODUCTION: Aquaporins (AQPs) are membrane proteins that facilitate water movement across biological membranes. Vaginal lubrication may be mediated by blood flow and other potential mechanisms related to transudation of fluid. The most common female sexual dysfunction in diabetes is inadequate vaginal lubrication. AIM: To investigate the expression of AQP1-3 in vaginal tissue of diabetes mellitus rats. METHODS: Female Sprague-Dawley rats (N = 20) were randomly divided into group A (12-week-old nondiabetic control, N = 5), group B (16-week-old nondiabetes control, N = 5), group C (12-week-old diabetes mellitus rats, N = 5), and group D (16-week-old diabetes mellitus rats, N = 5). Vaginal fluid was measured by fluid weight absorbed by cotton swabs after pelvic nerve electrostimulation and anterior vaginal tissue was dissected for determining the expression of AQP1-3 by immunohistochemical study and Western blot. MAIN OUTCOME MEASURES: The expression of AQP1-3 was determined in the vagina of diabetes mellitus rats by Western blot. RESULTS: There are no significant differences in serum estradiol concentrations of rats among these groups (P > 0.05). Vaginal fluid was significantly lower in group C (2.7 ± 0.67 mg) and group D (2.5 ± 1.03 mg) than in group A (5.74 ± 1.23 mg) and group B (5.5 ± 1.08 mg) (P < 0.05), respectively. The protein expressions of AQP1-3 were significantly lower in group C (43.40 ± 4.83, 60.60 ± 12.80, and 59.60 ± 6.95) and group D (20.81 ± 2.86, 47.80 ± 11.43, and 54.20 ± 5.26) than in group A (116.62 ± 3.21, 110.81 ± 8.044, and 108.80 ± 4.97) and group B (122.12 ± 14.54, 111.21 ± 15.07, and 106.40 ± 4.16) (P < 0.05), respectively. CONCLUSIONS: Decreased vaginal fluid in diabetes mellitus rats after electrostimulation may be partly due to estrogen-independent decreases of AQP1-3 in vaginal tissue.


Asunto(s)
Acuaporina 1/análisis , Acuaporina 2/análisis , Acuaporina 3/análisis , Diabetes Mellitus Experimental/patología , Vagina/patología , Animales , Western Blotting , Estimulación Eléctrica , Femenino , Técnicas para Inmunoenzimas , Ratas , Ratas Sprague-Dawley , Vagina/metabolismo
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