RESUMEN
Pulmonary arterial hypertension (PAH) is a chronic disease characterized by a progressive increase in mean pulmonary arterial pressure. Mutations in the BMPR2 and AQP1 genes have been described in familial PAH. The bone morphogenetic proteins BMP9 and BMP10 bind with high affinity to BMPR2. Administration of BMP9 has been proposed as a potential therapeutic strategy against PAH, although recent conflicting evidence dispute the effect of such a practice. Considering the involvement of the above molecules in PAH onset, progression, and therapeutic value, we examined the effects of modulation of BMP9, BMPR2, and AQP1 on BMP9, BMP10, BMPR2, AQP1, and TGFB1 expression in human pulmonary microvascular endothelial cells in vitro. Our results demonstrated that silencing the BMPR2 gene resulted in increased expression of its two main ligands, namely BMP9 and BMP10. Exogenous administration of BMP9 caused the return of BMP10 to basal levels, while it restored the decreased AQP1 protein levels and the decreased TGFB1 mRNA and protein expression levels caused by BMPR2 silencing. Moreover, AQP1 gene silencing also resulted in increased expression of BMP9 and BMP10. Our results might possibly imply that the effect of exogenously administered BMP9 on molecules participating in the BMP signaling pathway could depend on the expression levels of BMPR2. Taken together, these results may provide insight into the highly complex interactions of the BMP signaling pathway.
Asunto(s)
Acuaporina 1 , Receptores de Proteínas Morfogenéticas Óseas de Tipo II , Células Endoteliales , Factor 2 de Diferenciación de Crecimiento , Transducción de Señal , Factor de Crecimiento Transformador beta1 , Humanos , Acuaporina 1/metabolismo , Acuaporina 1/genética , Factor 2 de Diferenciación de Crecimiento/metabolismo , Factor 2 de Diferenciación de Crecimiento/genética , Células Endoteliales/metabolismo , Receptores de Proteínas Morfogenéticas Óseas de Tipo II/metabolismo , Receptores de Proteínas Morfogenéticas Óseas de Tipo II/genética , Factor de Crecimiento Transformador beta1/metabolismo , Pulmón/metabolismo , Pulmón/irrigación sanguínea , Microvasos/metabolismo , Microvasos/citología , Células Cultivadas , Silenciador del Gen , Hipertensión Arterial Pulmonar/metabolismo , Hipertensión Arterial Pulmonar/genética , Proteínas Morfogenéticas ÓseasRESUMEN
Nephron loop-vessel countercurrent arrangement in the medulla provides the structural basis for the formation of concentrated urine. To date, the morphogenesis of it and relevant water and solutes transportation has not been fully elucidated. In this study, with immunohistochemistry for aquaporins (AQP) and Na-K-2Cl co-transporter (NKCC2), as well as 3D visualization, we noticed in embryonic day 14.5 kidneys that the countercurrent arrangement of two pairs of loop-vessel was established as soon as the loop and vessel both extended into the medulla. One pair happened between descending limb and ascending vasa recta, the other occurred between thick ascending limb and descending vasa recta. Meanwhile, the immunohistochemical results showed that the limb and vessel expressing AQP-1 such as descending thick and thin limb and descending vasa recta was always accompanied with AQP-1 negative ascending vasa recta or capillaries and thick ascending limb, respectively. Moreover, the thick ascending limb expressing NKCC2 closely contacted with descending vasa recta without expressing NKCC2. As kidney developed, an increasing number of loop-vessels in countercurrent arrangement extended into the interstitium of the medulla. In addition, we observed that the AQP-2 positive ureteric bud and their branches were separated from those pairs of tubule-vessels by a relatively large and thin-walled veins or capillaries. Thus, the present study reveals that the loop-vessel countercurrent arrangement is formed at the early stage of nephrogenesis, which facilitates the efficient transportation of water and electrolytes to maintain the medullary osmolality and to form a concentrated urine.
Asunto(s)
Acuaporina 1 , Inmunohistoquímica , Miembro 1 de la Familia de Transportadores de Soluto 12 , Animales , Ratones , Miembro 1 de la Familia de Transportadores de Soluto 12/metabolismo , Acuaporina 1/metabolismo , Imagenología Tridimensional/métodos , Riñón/metabolismo , Riñón/embriología , Túbulos Renales/metabolismo , Asa de la Nefrona/metabolismo , Asa de la Nefrona/embriología , Acuaporinas/metabolismo , Nefronas/metabolismo , Nefronas/embriología , FemeninoRESUMEN
Acute respiratory infections (ARIs) are associated with high mortality and morbidity. Acute lung injury (ALI) is caused by the activation of immune cells during ARIs caused by viruses such as SARS-CoV-2. Aquaporin 1 (AQP1) is distributed in a variety of immune cells and is related to the occurrence of ALI, but the mechanism is not clear. A reference map of human single cells was used to identify macrophages in COVID-19 patients at the single-cell level. "FindMarkers" was used to analyze differentially expressed genes (DEGs), and "clusterProfiler" was used to analyze the functions of the DEGs. An M1 macrophage polarization model was established with lipopolysaccharide (LPS) in vitro, and the relationships among AQP1, pyroptosis and M1 polarization were examined by using an AQP1 inhibitor. Transcriptome sequencing and RT-qPCR were used to examine the molecular mechanism by which AQP1 regulates macrophage polarization and pyroptosis. Antigen presentation, M1 polarization, migration and phagocytosis are abnormal in SARS-CoV-2-infected macrophages, which is related to the high expression of AQP1. An M1 polarization model of macrophages was constructed in vitro, and an AQP1 inhibitor was used to examine whether AQP1 could promote M1 polarization and pyroptosis in response to LPS. Transcriptome and cell experiments showed that this effect was related to a decrease in chemokines caused by AQP1 deficiency. AQP1 participates in M1 polarization and pyroptosis in macrophages by increasing the levels of chemokines induced by LPS, which provides new insights for the diagnosis and treatment of ALI.
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Acuaporina 1 , COVID-19 , Macrófagos , Piroptosis , Animales , Humanos , Ratones , Acuaporina 1/metabolismo , Acuaporina 1/genética , COVID-19/inmunología , Lipopolisacáridos , Activación de Macrófagos , Macrófagos/metabolismo , Macrófagos/inmunología , SARS-CoV-2RESUMEN
Glaucoma affects approximately 80 million individuals worldwide, a condition for which current treatment options are inadequate. The primary risk factor for glaucoma is elevated intraocular pressure. Intraocular pressure is determined by the balance between the secretion and outflow of aqueous humor. Here we show that using the RNA interference tool CasRx based on shH10 adenovirus-associated virus can reduce the expression of the aqueous humor circulation related genes Rock1 and Rock2, as well as aquaporin 1 and ß2 adrenergic receptor in female mice. This significantly reduced intraocular pressure in female mice and provided protection to the retina ganglion cells, ultimately delaying disease progression. In addition, we elucidated the mechanisms by which the knockdown of Rock1 and Rock2, or aquaporin 1 and ß2 adrenergic receptor in female mice, reduces the intraocular pressure and secures the retina ganglion cells by single-cell sequencing.
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Acuaporina 1 , Sistemas CRISPR-Cas , Glaucoma , Presión Intraocular , Células Ganglionares de la Retina , Quinasas Asociadas a rho , Animales , Células Ganglionares de la Retina/metabolismo , Células Ganglionares de la Retina/patología , Quinasas Asociadas a rho/metabolismo , Quinasas Asociadas a rho/genética , Femenino , Acuaporina 1/metabolismo , Acuaporina 1/genética , Ratones , Glaucoma/genética , Glaucoma/metabolismo , Humor Acuoso/metabolismo , Ratones Endogámicos C57BL , Modelos Animales de Enfermedad , Interferencia de ARN , HumanosRESUMEN
Genetically encoded reporters for magnetic resonance imaging (MRI) offer a valuable technology for making molecular-scale measurements of biological processes within living organisms with high anatomical resolution and whole-organ coverage without relying on ionizing radiation. However, most MRI reporters rely on synthetic contrast agents, typically paramagnetic metals and metal complexes, which often need to be supplemented exogenously to create optimal contrast. To eliminate the need for synthetic contrast agents, we previously introduced aquaporin-1, a mammalian water channel, as a new reporter gene for the fully autonomous detection of genetically labeled cells using diffusion-weighted MRI. In this study, we aimed to expand the toolbox of diffusion-based genetic reporters by modulating aquaporin membrane trafficking and harnessing the evolutionary diversity of water channels across species. We identified a number of new water channels that functioned as diffusion-weighted reporter genes. In addition, we show that loss-of-function variants of yeast and human aquaporins can be leveraged to design first-in-class diffusion-based sensors for detecting the activity of a model protease within living cells.
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Técnicas Biosensibles , Imagen de Difusión por Resonancia Magnética , Genes Reporteros , Imagen de Difusión por Resonancia Magnética/métodos , Humanos , Técnicas Biosensibles/métodos , Acuaporina 1/genética , Agua/química , Acuaporinas/genética , Acuaporinas/metabolismoRESUMEN
Di-2-ethylhexyl phthalate (DEHP) is frequently used as a plasticizer to enhance the plasticity and durability of agricultural products, which pose adverse effects to human health and the environment. Aquaporin 1 (AQP1) is a main water transport channel protein and is involved in the maintenance of intestinal integrity. However, the impact of DEHP exposure on gut health and its potential mechanisms remain elusive. Here, we determined that DEHP exposure induced a compromised duodenum structure, which was concomitant with mitochondrial structural injury of epithelial cells. Importantly, DEHP exposure caused duodenum inflammatory epithelial cell damage and strong inflammatory response accompanied by activating the TLR4/MyD88/NF-κB signaling pathway. Mechanistically, DEHP exposure directly inhibits the expression of AQP1 and thus leads to an inflammatory response, ultimately disrupting duodenum integrity and barrier function. Collectively, our findings uncover the role of AQP1 in phthalate-induced intestinal disorders, and AQP1 could be a promising therapeutic approach for treating patients with intestinal disorders or inflammatory diseases.
Asunto(s)
Acuaporina 1 , Mucosa Intestinal , Animales , Acuaporina 1/genética , Acuaporina 1/metabolismo , Ratones , Humanos , Mucosa Intestinal/metabolismo , Mucosa Intestinal/efectos de los fármacos , Ratones Endogámicos C57BL , Inflamación/metabolismo , Inflamación/genética , Inflamación/inducido químicamente , Masculino , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , FN-kappa B/metabolismo , FN-kappa B/genética , Dietilhexil Ftalato/toxicidad , Ácidos Ftálicos , Transducción de Señal/efectos de los fármacosRESUMEN
Despite continuous medical advancements, traumatic brain injury (TBI) remains a leading cause of death and disability worldwide. Consequently, there is a pursuit for biomarkers that allow non-invasive monitoring of patients after cranial trauma, potentially improving clinical management and reducing complications and mortality. Aquaporins (AQPs), which are crucial for transmembrane water transport, may be significant in this context. This study included 48 patients, with 27 having acute (aSDH) and 21 having chronic subdural hematoma (cSDH). Blood plasma samples were collected from the participants at three intervals: the first sample before surgery, the second at 15 h, and the third at 30 h post-surgery. Plasma concentrations of AQP1, AQP2, AQP4, and AQP9 were determined using the sandwich ELISA technique. CT scans were performed on all patients pre- and post-surgery. Correlations between variables were examined using Spearman's nonparametric rank correlation coefficient. A strong correlation was found between aquaporin 2 levels and the volume of chronic subdural hematoma and midline shift. However, no significant link was found between aquaporin levels (AQP1, AQP2, AQP4, and AQP9) before and after surgery for acute subdural hematoma, nor for AQP1, AQP4, and AQP9 after surgery for chronic subdural hematoma. In the chronic SDH group, AQP2 plasma concentration negatively correlated with the midline shift measured before surgery (Spearman's ρ -0.54; p = 0.017) and positively with hematoma volume change between baseline and 30 h post-surgery (Spearman's ρ 0.627; p = 0.007). No statistically significant correlation was found between aquaporin plasma levels and hematoma volume for AQP1, AQP2, AQP4, and AQP9 in patients with acute SDH. There is a correlation between chronic subdural hematoma volume, measured radiologically, and serum AQP2 concentration, highlighting aquaporins' potential as clinical biomarkers.
Asunto(s)
Acuaporina 2 , Biomarcadores , Edema Encefálico , Humanos , Masculino , Femenino , Biomarcadores/sangre , Persona de Mediana Edad , Anciano , Pronóstico , Edema Encefálico/sangre , Edema Encefálico/etiología , Edema Encefálico/diagnóstico por imagen , Acuaporina 2/sangre , Acuaporina 2/metabolismo , Adulto , Traumatismos Craneocerebrales/sangre , Traumatismos Craneocerebrales/complicaciones , Hematoma Subdural Crónico/sangre , Hematoma Subdural Crónico/cirugía , Acuaporina 1/sangre , Acuaporina 1/metabolismo , Tomografía Computarizada por Rayos X , Lesiones Traumáticas del Encéfalo/sangre , Lesiones Traumáticas del Encéfalo/diagnóstico , Acuaporinas/sangre , Acuaporinas/metabolismoRESUMEN
Leptin is an obesity-related hormone that plays an important role in breast cancer progression. Vasculogenic mimicry (VM) refers to the formation of vascular channels lined by tumor cells. This study aimed to investigate the relationship between leptin and VM in human breast cancer cells. VM was measured by a 3D culture assay. Signal transducers and activators of transcription 3 (STAT3) signaling, aquaporin-1 (AQP1), and the expression of VM-related proteins, including vascular endothelial cadherin (VE-cadherin), twist, matrix metalloproteinase-2 (MMP-2), and laminin subunit 5 gamma-2 (LAMC2), were examined by Western blot. AQP1 mRNA was analyzed by a reverse transcriptase-polymerase chain reaction (RT-PCR). Leptin increased VM and upregulated phospho-STAT3, VE-cadherin, twist, MMP-2, and LAMC2. These effects were inhibited by the leptin receptor-blocking peptide, Ob-R BP, and the STAT3 inhibitor, AG490. A positive correlation between leptin and AQP1 mRNA was observed and was confirmed by RT-PCR. Leptin upregulated AQP1 expression, which was blocked by Ob-R BP and AG490. AQP1 overexpression increased VM and the expression of VM-related proteins. AQP1 silencing inhibited leptin-induced VM and the expression of VM-related proteins. Thus, these results showed that leptin facilitates VM in breast cancer cells via the Ob-R/STAT3 pathway and that AQP1 is a key mediator in leptin-induced VM.
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Neoplasias de la Mama , Leptina , Neovascularización Patológica , Femenino , Humanos , Antígenos CD , Acuaporina 1/metabolismo , Acuaporina 1/genética , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Cadherinas/metabolismo , Cadherinas/genética , Línea Celular Tumoral , Regulación Neoplásica de la Expresión Génica , Laminina/metabolismo , Leptina/metabolismo , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 2 de la Matriz/genética , Células MCF-7 , Neovascularización Patológica/metabolismo , Neovascularización Patológica/genética , Transducción de Señal , Factor de Transcripción STAT3/metabolismo , Factor de Transcripción STAT3/genéticaRESUMEN
Understanding how the amino acid sequence dictates protein structure and defines its stability is a fundamental problem in molecular biology. It is especially challenging for membrane proteins that reside in the complex environment of a lipid bilayer. Here, we obtain an atomic-level picture of the thermally induced unfolding of a membrane-embedded α-helical protein, human aquaporin 1, using solid-state nuclear magnetic resonance spectroscopy. Our data reveal the hierarchical two-step pathway that begins with unfolding of a structured extracellular loop and proceeds to an intermediate state with a native-like helical packing. In the second step, the transmembrane domain unravels as a single unit, resulting in a heterogeneous misfolded state with high helical content but with nonnative helical packing. Our results show the importance of loops for the kinetic stabilization of the whole membrane protein structure and support the three-stage membrane protein folding model.
Asunto(s)
Proteínas de la Membrana , Desplegamiento Proteico , Humanos , Proteínas de la Membrana/química , Proteínas de la Membrana/metabolismo , Acuaporina 1/química , Acuaporina 1/metabolismo , Resonancia Magnética Nuclear Biomolecular , Espectroscopía de Resonancia Magnética/métodos , Modelos Moleculares , Pliegue de Proteína , Cinética , TermodinámicaRESUMEN
AIMS: In patients with diabetic microvascular complications, decreased perfusion or vascular occlusion, caused by reduced vascular diameter, is a common characteristic that will lead to insufficient blood supply. Yet, the regulatory mechanism and effective treatment approach remain elusive. METHODS AND RESULTS: Our initial findings revealed a notable decrease in the expression of human AQP1 in both diabetic human retina samples (49 healthy vs. 54 diabetic samples) and high-glucose-treated human retinal microvascular endothelial cells. Subsequently, our investigations unveiled a reduction in vascular diameter and compromised perfusion within zebrafish embryos subjected to high glucose treatment. Further analysis indicated a significant down-regulation of two aquaporins, aqp1a.1 and aqp8a.1, which are highly enriched in ECs and are notably responsive to hyperglycaemic conditions. Intriguingly, the loss of function of aqp1a.1 and/or aqp8a.1 resulted in a reduction of intersegmental vessel diameters, effectively mirroring the phenotype observed in the hyperglycaemic zebrafish model. The overexpression of aqp1a.1/aqp8a.1 in zebrafish ECs led to notable enlargement of microvascular diameters. Moreover, the reduced vessel diameters resulting from high-glucose treatment were effectively rescued by the overexpression of these aquaporins. Additionally, both aqp1a.1 and apq8a.1 were localized in the intracellular vacuoles in cultured ECs as well as the ECs of sprouting ISVs, and the loss of Aqps caused the reduction of those vacuoles, which was required for lumenization. Notably, while the loss of AQP1 did not impact EC differentiation from human stem cells, it significantly inhibited vascular formation in differentiated ECs. CONCLUSION: EC-enriched aquaporins regulate the diameter of blood vessels through an intracellular vacuole-mediated process under hyperglycaemic conditions. These findings collectively suggest that aquaporins expressed in ECs hold significant promise as potential targets for gene therapy aimed at addressing vascular perfusion defects associated with diabetes.
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Acuaporina 1 , Acuaporinas , Células Endoteliales , Hiperglucemia , Microvasos , Vasos Retinianos , Vacuolas , Proteínas de Pez Cebra , Pez Cebra , Pez Cebra/metabolismo , Animales , Humanos , Acuaporina 1/metabolismo , Acuaporina 1/genética , Vacuolas/metabolismo , Vacuolas/patología , Células Endoteliales/metabolismo , Células Endoteliales/patología , Acuaporinas/metabolismo , Acuaporinas/genética , Hiperglucemia/metabolismo , Hiperglucemia/genética , Hiperglucemia/fisiopatología , Microvasos/metabolismo , Microvasos/patología , Vasos Retinianos/metabolismo , Vasos Retinianos/patología , Vasos Retinianos/fisiopatología , Proteínas de Pez Cebra/metabolismo , Proteínas de Pez Cebra/genética , Células Cultivadas , Retinopatía Diabética/metabolismo , Retinopatía Diabética/patología , Retinopatía Diabética/genética , Retinopatía Diabética/fisiopatología , Estudios de Casos y Controles , Animales Modificados Genéticamente , Glucemia/metabolismoRESUMEN
BACKGROUND: Preventing joint edema is crucial in halting osteoarthritis (OA) progression. Growing clinical evidence indicate that Jianpi-Tongluo Formula (JTF) may have a promising anti-edema effect. However, the therapeutic properties of JTF and the underlying mechanisms remains unclear. MATERIALS AND METHODS: An OA rat model was established and employed to evaluate pharmacological effects of JTF in vivo based on dynamic histopathologic assessments and micro-CT observations. Then, OA-related genes and potential targets of JTF were identified through clinical transcriptomic data analysis and "disease gene-drug target" network analysis, which were verified by a series of in vivo experiments. RESULTS: JTF administration effectively reduced pain and joint edema, inhibited matrix degradation, chondrocyte apoptosis, and aquaporin expression in OA rats. Notably, JTF dose-dependently reversed damage-associated molecular patterns and inflammatory factor upregulation. Mechanically, our "disease gene-drug target" network analysis indicated that the NCOA4-HMGB1-GSK3B-AQPs axis, implicated in ferroptosis and aquaporin dysregulation, may be potentially served as a target of JTF against OA. Accordingly, JTF mitigated NCOA4, HMGB1, and GSK3B expression, oxidative stress, and iron metabolism aberrations in OA rats. Furthermore, JTF treatment significantly attenuated the aberrant upregulation of AQP1, AQP3, and AQP4 proteins observed in cartilage tissues of OA rats. CONCLUSION: Our data reveal for the first time that JTF may exert cartilage protective and anti-edema effects in osteoarthritis therapy by inhibiting NCOA4-HMGB1-driven ferroptosis and aquaporin dysregulation.
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Ferroptosis , Proteína HMGB1 , Osteoartritis , Ratas Sprague-Dawley , Animales , Osteoartritis/tratamiento farmacológico , Osteoartritis/metabolismo , Ferroptosis/efectos de los fármacos , Ratas , Masculino , Proteína HMGB1/metabolismo , Medicamentos Herbarios Chinos/farmacología , Edema/tratamiento farmacológico , Acuaporinas/metabolismo , Coactivadores de Receptor Nuclear/metabolismo , Modelos Animales de Enfermedad , Acuaporina 3/metabolismo , Acuaporina 1/metabolismoRESUMEN
The development of genetic reporters for magnetic resonance imaging (MRI) is essential for investigating biological functions inâ vivo. However, current MRI reporters have low sensitivity, making it challenging to create significant contrast against the tissue background, especially when only a small fraction of cells express the reporter. To overcome this limitation, we developed an approach for amplifying the sensitivity of molecular MRI by combining a chemogenetic contrast mechanism with a biophysical approach to increase water diffusion through the co-expression of a dual-gene construct comprising an organic anion transporting polypeptide, Oatp1b3, and a water channel, Aqp1. We first show that the expression of Aqp1 amplifies MRI contrast in cultured cells engineered to express Oatp1b3. We demonstrate that the contrast amplification is caused by Aqp1-driven increase in water exchange, which provides the gadolinium ions internalized by Oatp1b3-expressing cells with access to a larger water pool compared with exchange-limited conditions. We further show that our methodology allows cells to be detected using approximately 10-fold lower concentrations of gadolinium than that in the Aqp1-free scenario. Finally, we show that our approach enables the imaging of mixed-cell cultures containing a low fraction of Oatp1b3-labeled cells that are undetectable on the basis of Oatp1b3 expression alone.
Asunto(s)
Acuaporina 1 , Genes Reporteros , Imagen por Resonancia Magnética , Miembro 1B3 de la Familia de los Transportadores de Solutos de Aniones Orgánicos , Agua , Agua/química , Humanos , Imagen por Resonancia Magnética/métodos , Acuaporina 1/metabolismo , Acuaporina 1/genética , Miembro 1B3 de la Familia de los Transportadores de Solutos de Aniones Orgánicos/metabolismo , Miembro 1B3 de la Familia de los Transportadores de Solutos de Aniones Orgánicos/genética , Gadolinio/química , Medios de Contraste/química , Medios de Contraste/metabolismo , Células HEK293 , AnimalesRESUMEN
Pulmonary hypertension (PH) is a condition in which remodeling of the pulmonary vasculature leads to hypertrophy of the muscular vascular wall and extension of muscle into nonmuscular arteries. These pathological changes are predominantly due to the abnormal proliferation and migration of pulmonary arterial smooth muscle cells (PASMCs), enhanced cellular functions that have been linked to increases in the cell membrane protein aquaporin 1 (AQP1). However, the mechanisms underlying the increased AQP1 abundance have not been fully elucidated. Here we present data that establishes a novel interaction between AQP1 and the proteolytic enzyme caspase-3. In silico analysis of the AQP1 protein reveals two caspase-3 cleavage sites on its C-terminal tail, proximal to known ubiquitin sites. Using biotin proximity ligase techniques, we establish that AQP1 and caspase-3 interact in both human embryonic kidney (HEK) 293A cells and rat PASMCs. Furthermore, we demonstrate that AQP1 levels increase and decrease with enhanced caspase-3 activity and inhibition, respectively. Ultimately, further work characterizing this interaction could provide the foundation for novel PH therapeutics.NEW & NOTEWORTHY Pulmonary arterial smooth muscle cells (PASMCs) are integral to pulmonary vascular remodeling, a characteristic of pulmonary arterial hypertension (PAH). PASMCs isolated from robust animal models of disease demonstrate enhanced proliferation and migration, pathological functions associated with increased abundance of the membrane protein aquaporin 1 (AQP1). We present evidence of a novel interaction between the proteolytic enzyme caspase-3 and AQP1, which may control AQP1 abundance. These data suggest a potential new target for novel PAH therapies.
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Acuaporina 1 , Caspasa 3 , Músculo Liso Vascular , Miocitos del Músculo Liso , Arteria Pulmonar , Animales , Humanos , Masculino , Ratas , Acuaporina 1/metabolismo , Acuaporina 1/genética , Caspasa 3/metabolismo , Proliferación Celular , Células HEK293 , Hipertensión Pulmonar/metabolismo , Hipertensión Pulmonar/patología , Músculo Liso Vascular/metabolismo , Músculo Liso Vascular/patología , Miocitos del Músculo Liso/metabolismo , Miocitos del Músculo Liso/patología , Arteria Pulmonar/metabolismo , Arteria Pulmonar/patología , Ratas Sprague-DawleyRESUMEN
BACKGROUND: Nephroblastoma, also known as Wilms' tumor (WT), is an embryonic malignant tumor and one of the most common malignant tumors in the abdominal region of children. The exact role and underlying mechanisms of aquaporin-1 (AQP1) in the occurrence and development of nephroblastoma remain unclear. METHODS: After overexpression of AQP1, cell proliferation was assessed using the CCK-8 proliferation assay and EdU staining. Flow cytometry was employed to assess cell apoptosis, and Western blotting (WB) analysis was conducted to validate the expression of relevant protein markers. mRNA sequencing (mRNA-Seq) was performed on WT cells overexpressing AQP1 to predict and characterize the associated mechanisms. Transmission electron microscopy was utilized to observe changes in the ultrastructure of WT cells undergoing apoptosis and pyroptosis following AQP1 overexpression. Functional in vivo validation was conducted through animal experiments. RESULTS: We validated that overexpression of AQP1 inhibited cell proliferation and promoted cell apoptosis and pyroptosis both in vitro and in vivo. mRNA-Seq analysis of WT cells with AQP1 overexpression suggested that these effects might be mediated through the inhibition of the JAK-STAT signaling pathway. Additionally, we discovered that overexpression of AQP1 activated the classical pyroptosis signaling pathway dependent on caspase-1, thereby promoting pyroptosis in WT. CONCLUSION: These findings highlight the important functional role of AQP1 in the pathobiology of nephroblastoma, providing novel insights into the development of this disease. Moreover, these results offer new perspectives on the potential therapeutic targeting of AQP1 as a treatment strategy for nephroblastoma.
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Acuaporina 1 , Neoplasias Renales , Tumor de Wilms , Animales , Humanos , Apoptosis , Línea Celular Tumoral , Proliferación Celular , Neoplasias Renales/genética , Neoplasias Renales/patología , Piroptosis/genética , ARN Mensajero/genética , Tumor de Wilms/genética , Tumor de Wilms/patología , Acuaporina 1/genéticaRESUMEN
BACKGROUND/AIM: Aquaporins (AQPs) were initially discovered as water channel proteins that facilitate transcellular water movements. Recent studies have shown that AQPs are expressed and play an oncogenic role in various cancers. However, the expression and role of Aquaporin 4 (AQP4) in colon cancer have not been investigated. This study aimed to examine the clinical and pathophysiologic significance of AQP4 in colon cancer. PATIENTS AND METHODS: Immunohistochemistry (IHC) of AQP4 for 145 primary tumor samples obtained from patients with stage II or III colon cancer was performed, and the relationship between AQP4 expression and patients' prognoses was analyzed. Knockdown experiments with AQP4 small interfering RNA using human colon cancer cells were conducted to analyze the effects on cell invasiveness. RESULTS: IHC revealed that AQP4 was scarcely expressed in the noncancerous colonic mucosa. Of the 145 patients who enrolled in this study, 109 (75.2%) and 36 (24.8%) patients were classified as negative and positive for AQP4 expression, respectively. A high level of AQP4 expression is significantly associated with deeper tumors with lymph node metastasis and venous invasion. A 5-year progression-free survival rate of AQP4-positive patients was significantly worse than that of AQP-4 negative patients (70.7% vs. 87.0%, p=0.049). Furthermore, AQP4 knockdown significantly inhibited cell migration and invasion in HCT116 cells. CONCLUSION: AQP4 may be a novel biomarker and therapeutic target for colon cancer.
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Acuaporina 4 , Neoplasias del Colon , Humanos , Acuaporina 4/genética , Acuaporina 4/metabolismo , ARN Interferente Pequeño/genética , Inmunohistoquímica , Neoplasias del Colon/genética , Acuaporina 1/genética , Acuaporina 1/metabolismoRESUMEN
Cancer invasion and metastasis are known to be potentiated by the expression of aquaporins (AQPs). Likewise, the expression levels of AQPs have been shown to be prognostic for survival in patients and have a role in tumor growth, edema, angiogenesis, and tumor cell migration. Thus, AQPs are key players in cancer biology and potential targets for drug development. Here, we present the single-particle cryo-EM structure of human AQP7 at 3.2-Å resolution in complex with the specific inhibitor compound Z433927330. The structure in combination with MD simulations shows that the inhibitor binds to the endofacial side of AQP7. In addition, cancer cells treated with Z433927330 show reduced proliferation. The data presented here serve as a framework for the development of AQP inhibitors.
Asunto(s)
Acuaporinas , Neoplasias , Humanos , Acuaporinas/metabolismo , Acuaporina 1/metabolismoRESUMEN
The goals of this study were to investigate whether Wnt/ß-catenin signaling plays a role in hypo-osmolality-related degeneration of nucleus pulposus (NP) cells, and if so, to define the mechanism underlying AQP1 in this effect. Human NP cells were cultured under hypo-osmotic (300/350/400 mOsm) and iso-osmotic (450 mOsm) conditions. The cell viability, AQP1, the expression of Wnt/ß-catenin signaling, collagen II/I, and MMP3/9 were evaluated. To determine the effects of the Wnt/ß-catenin signaling, we used the inhibitor and the activator of Wnt during the hypo-osmotic culture of NP cells. We also examined whether the silencing and overexpressing of the AQP1 gene would affect the Wnt/ß-catenin expression in NP cells. Hypo-osmolality caused NP cell degeneration and activated the Wnt/ß-catenin signaling but suppressed the AQP1 level. Inhibiting the Wnt/ß-catenin signaling alleviated the hypo-osmolality-induced NP cell degeneration. On the contrary, activating Wnt/ß-catenin aggravated the NP cell degeneration under hypo-osmotic conditions, which did not affect AQP1 expression. AQP1-overexpressed NP cells exhibited decreased Wnt/ß-catenin signaling and alleviated cell degeneration under the hypo-osmotic condition. Besides, AQP1 silencing accelerated NP cell degeneration and activated Wnt/ß-catenin expression compared with untreated control. Hypo-osmolality promotes NP cell degeneration via activating Wnt/ß-catenin signaling, which is suppressed by AQP1 expression. The upregulation of AQP1 suppressed the Wnt/ß-catenin signaling and alleviated the hypo-osmolality induced by the NP cell degeneration.
Asunto(s)
Degeneración del Disco Intervertebral , Núcleo Pulposo , Humanos , Núcleo Pulposo/metabolismo , Degeneración del Disco Intervertebral/genética , Degeneración del Disco Intervertebral/metabolismo , beta Catenina/genética , beta Catenina/metabolismo , Células Cultivadas , Vía de Señalización Wnt/fisiología , Acuaporina 1/genética , Acuaporina 1/metabolismoRESUMEN
Aquaporins (AQPs) are integral membrane proteins that act as water channels for which a total of 13 orthologs of AQP genes in birds have been reported. Tissue expression and cellular or subcellular localization of AQPs have been poorly investigated in the male reproductive system of birds. We aimed to determine the distribution and localization of AQP5 and AQP7 proteins by immunocytochemistry in testicular tissues obtained from developing chicks (14, 21, 28, 35 and 42 days old). Totally 175 male chicks (Ross 308) were used in the study from which testicular tissue was removed, fixed in 10% formaldehyde solution, then embedded in paraffin blocks. Five µm sections were cut, mounted on poly-L-lysine slides, dried in an oven, then dehydrated using standard immunohistochemistry staining protocol. The sections were imaged with a Nikon Eclipse 50i trinocular light microscope. Immunohistochemical evaluation of the immune reactivity of AQP5 revealed a positive immune reaction in spermatocytes and interstitial areas of the testes in 14-day-old chicks. Testicular tissue AQP5 immune reactivity was observed in the tubule and the interstitial regions of 21-, 28-, 35- and 42-day-old chicks. AQP7 immune reactions were determined in the tubule and interstitial areas testes of developing chicks' testis tissue, with increasing positivity corresponding to older age. The expression of AQP5 and AQP7 appears to be species-specific due to differences in localization and expression in male chicks compared with studies of other mammals, which is likely to play an important role in regulating fluid and sperm volume. This research can serve as a base for future studies that will contribute to the understanding of the male genital system of AQPs.
Asunto(s)
Acuaporina 5 , Testículo , Masculino , Animales , Testículo/metabolismo , Acuaporina 5/metabolismo , Semen , Espermatozoides , Pollos , Acuaporina 1/metabolismo , MamíferosAsunto(s)
Acuaporina 1 , Neoplasias , Humanos , Acuaporina 1/genética , Neoplasias/genética , Genes Supresores de Tumor , OncogenesRESUMEN
Rheumatoid arthritis (RA) is an autoimmune inflammatory disease affecting approximately 1% of the global population, with a higher prevalence in women than in men. Chronic inflammation and oxidative stress play pivotal roles in the pathogenesis of RA. Anethole, a prominent compound derived from fennel (Foeniculum vulgare), possesses a spectrum of therapeutic properties, including anti-arthritic, anti-inflammatory, antioxidant, and tumor-suppressive effects. However, its specific impact on RA remains underexplored. This study sought to uncover the potential therapeutic value of anethole in treating RA by employing an H2 O2 -induced inflammation model with HIG-82 synovial cells. Our results demonstrated that exposure to H2 O2 induced the inflammation and apoptosis in these cells. Remarkably, anethole treatment effectively countered these inflammatory and apoptotic processes triggered by H2 O2 . Moreover, we identified the aquaporin 1 (AQP1) and protein kinase A (PKA) pathway as critical regulators of inflammation and apoptosis. H2 O2 stimulation led to an increase in the AQP1 expression and a decrease in p-PKA-C, contributing to cartilage degradation. Conversely, anethole not only downregulated the AQP1 expression but also activated the PKA pathway, effectively suppressing cell inflammation and apoptosis. Furthermore, anethole also inhibited the enzymes responsible for cartilage degradation. In summary, our findings highlight the potential of anethole as a therapeutic agent for mitigating H2 O2 -induced inflammation and apoptosis in synovial cells, offering promising prospects for future RA treatments.