Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 99
Filter
1.
Braz. j. med. biol. res ; 54(6): e10474, 2021. tab, graf
Article in English | LILACS | ID: biblio-1285667

ABSTRACT

Osteosarcoma is a highly malignant tumor that occurs in the bone. Previous studies have shown that multiple microRNAs (miRNAs) regulate the development of osteosarcoma. This study aimed to explore the role of miR-629-5p and its target gene, caveolin 1 (CAV1), in osteosarcoma development. To analyze the expression of miR-629-5p and CAV1 mRNA in osteosarcoma tissues and cell lines, qRT-PCR analysis was performed. Dual-luciferase reporter experiments were subsequently performed to validate the relationship between CAV1 and miR-629-5p. CCK8 assay was used to measure osteosarcoma cell proliferation, and wound-healing assay was performed to study their migratory phenotype. Our findings revealed that miR-629-5p was overexpressed in osteosarcoma tissues and cells, and thereby enhanced cell proliferation and migration. Further, we validated that miR-629-5p targets CAV1 mRNA directly. CAV1 expression, which was negatively correlated with miR-629-5p expression, was found to be downregulated in osteosarcoma tissue samples. Moreover, our data showed that an increase in CAV1 level led to a decline in osteosarcoma cell proliferation and migration, which could be rescued by miR-629-5p upregulation. Overall, our study confirmed that miR-629-5p promoted osteosarcoma proliferation and migration by directly inhibiting CAV1.


Subject(s)
Humans , Bone Neoplasms/genetics , Osteosarcoma/genetics , MicroRNAs/genetics , Gene Expression Regulation, Neoplastic , Cell Movement/genetics , Cell Line, Tumor , Cell Proliferation/genetics , Caveolin 1/genetics
2.
Braz. j. med. biol. res ; 54(10): e10653, 2021. tab, graf
Article in English | LILACS | ID: biblio-1285657

ABSTRACT

Vasculogenic mimicry (VM) plays an important role in human glioma progression and resistance to antiangiogenic therapy as a compensatory neovascularization mechanism in malignant tumors. Caveolin-1 (Cav-1) has been found to contribute to VM formation. However, it remains largely unknown whether Cav-1 expression correlates with VM in glioma. In this study, we examined CAV-1 expression levels and VM in human glioma cell lines and in 94 human gliomas with different grades of malignancy, and present Cox proportional hazards regression. The molecular role of Cav-1 in glioma cells was investigated using quantitative polymerase chain reaction (qRT-PCR) assays, western blotting, CCK-8 assays, and tubule formation assays. Cav-1 expression and VM formation were positively correlated with each other and both were closely associated with glioma development and progression as evidenced by the presence of cystic tumor, shortened survival time, and advanced-stage glioma in glioma patients with Cav-1 overexpression/increased VM formation. Cav-1 promoted U251 glioma cell proliferation and VM formation in a Matrigel-based 3D culture model. VM-associated factors including hypoxia-inducible factor 1α (HIF-1α) and p-Akt was significantly elevated by Cav-1 overexpression but suppressed by siCav-1 in U251 cells. Collectively, our study identified Cav-1 as an important regulator of glioma cell proliferation and VM formation, contributing to glioma development and progression.


Subject(s)
Humans , Caveolin 1/genetics , Glioma , Cell Line, Tumor , Cell Proliferation , Neovascularization, Pathologic
3.
Article in Korean | WPRIM | ID: wpr-811256

ABSTRACT

PURPOSE: Diabetic nephropathy is one of the most important diabetic complications prompted by chronic hyperglycemia, characterized by glomerulosclerosis, tubular fibrosis, and it eventually causes kidney failure. Nobiletin is a polymethoxyflavone present in tangerine and other citrus peels, and has anti-cancer and anti-inflammatory effects. This study investigated the effects of nobiletin on glomerular fibrosis through inhibition of the transforming growth factor (TGF)-β1-Src-caveolin-1 pathway.METHODS: Human renal mesangial cells (HRMC) were incubated in media containing 33 mM glucose with or without 1–20 uM nobiletin for 3 day. The cellular expression levels of fibrogenic collagen IV, fibronectin, connective tissue growth factor (CTGF), TGF-β1, Src and caveolin-1 were all examined. In addition, TGF-β1, Src and caveolin-1 proteins were screened to reveal the relationship among TGF-β1-Src-caveolin-1 signaling in glomerular fibrosis.RESULTS: High glucose promoted the production of collagen IV, fibronectin and CTGF in HRMC, which was inhibited in a dose dependent manner by 1–20 uM nobiletin. The Western blot data showed that high glucose elevated the expression of TGF-β1, Src, caveolin-1 and Rho GTPase. When nobiletin was treated to the HRMC exposed to high glucose, the expression of TGF-β1-Src-caveolin-1 was dampened. Finally, TGF-β1-Src-caveolin-1 signaling pathway was activated in high glucose-exposed HRMC, and such activation was encumbered by nobiletin.CONCLUSION: These result demonstrated that nobiletin blunted high glucose-induced extracellular matrix accumulation via inhibition of the TGF-β1-Src-caveolin-1 related intracellular signaling pathway. Nobiletin may be a potent renoprotective agent to counteract diabetes-associated glomerular fibrosis that leads to kidney failure.


Subject(s)
Blotting, Western , Caveolin 1 , Citrus , Collagen , Connective Tissue Growth Factor , Diabetes Complications , Diabetic Nephropathies , Extracellular Matrix , Fibronectins , Fibrosis , Glucose , GTP Phosphohydrolases , Humans , Hyperglycemia , Mesangial Cells , Renal Insufficiency , Transforming Growth Factors
4.
Article in Chinese | WPRIM | ID: wpr-828042

ABSTRACT

To explore whether paeonol can play an anti-atherosclerotic role by regulating the expression of aortic caveolin-1 and affecting NF-κB pathway, so as to inhibit the inflammatory response of vascular endothelium in atherosclerotic rats. The atherosclerotic model of rats was induced by high-fat diet and vitamin D_2. The primary culture of vascular endothelial cells(VECs) was carried out by tissue block pre-digestion and adherent method. The injury model of VECs was induced by lipopolysaccharide(LPS), and filipin, a small concave protein inhibitor, was added for control. HE staining was used to observe pathological changes of aorta. TNF-α, IL-6 and VCAM-1 were detected by ELISA. Western blot assay was used to detect the protein expression levels of caveolin-1 and p65 in aorta and VECs. The results showed that as compared with model group, paeonol significantly reduced aortic plaque area and lesion degree in rats, decreased the level of serum TNF-α, IL-6 and VCAM-1 in the rats and enhanced the relative expression level of caveolin-1, decreased p65 expression conversely(P<0.05 or P<0.01). In vitro, as compared to model group, paeonol obviously improved cell morphology, decreased the secretion of TNF-α, IL-6 and VCAM-1 in VECs, increased caveolin-1 expression, and decreased p65 protein expression(P<0.05 or P<0.01). Furthermore, filipin could reverse the effect of paeonol on expression of inflammatory factors and proteins(P<0.05 or P<0.01). According to the results, it was found that paeonol could play the role of anti-atherosclerosis by up-regulating the expression of caveolin-1 and inhibiting the activation of NF-κB pathway to reduce vascular inflammation in atherosclerotic rats.


Subject(s)
Acetophenones , Animals , Caveolin 1 , Endothelial Cells , Endothelium, Vascular , Inflammation , NF-kappa B , Rats , Signal Transduction , Tumor Necrosis Factor-alpha , Up-Regulation
5.
Article in English | WPRIM | ID: wpr-763711

ABSTRACT

BACKGROUND: Striatin and caveolin-1 (cav-1) are scaffolding/regulating proteins that are associated with salt-sensitive high blood pressure and promote renal sodium and water reabsorption, respectively. The mineralocorticoid receptor (MR) interacts with striatin and cav-1, while aldosterone increases striatin and cav-1 levels. However, no in vivo data have been reported for the levels of these proteins in the kidney. METHODS: Male Wistar rats were intraperitoneally injected with normal saline solution, aldosterone alone (Aldo: 150 µg/kg body weight), or aldosterone after pretreatment with eplerenone, an MR blocker, 30 minutes before the aldosterone injection (eplerenone [Ep.]+Aldo). Thirty minutes after the aldosterone injection, the amount and localization of striatin and cav-1 were determined by Western blot analysis and immunohistochemistry, respectively. RESULTS: Aldosterone increased striatin levels by 150% (P<0.05), and cav-1 levels by 200% (P<0.001). Eplerenone had no significant effect on striatin levels, but partially blocked the aldosterone-induced increase in cav-1 levels. Aldosterone stimulated striatin and cav-1 immunoreactivity in both the cortex and medulla. Eplerenone reduced cav-1 immunostaining in both areas; however, striatin intensity was reduced in the cortex, but increased in the medulla. CONCLUSION: This is the first in vivo study demonstrating that aldosterone rapidly enhances renal levels of striatin and cav-1. Aldosterone increases striatin levels via an MR-independent pathway, whereas cav-1 is partially regulated through MR.


Subject(s)
Aldosterone , Animals , Blotting, Western , Caveolin 1 , Humans , Hypertension , Immunohistochemistry , Kidney , Male , Rats , Rats, Wistar , Receptors, Mineralocorticoid , Sodium , Sodium Chloride , Water
6.
Araçatuba; s.n; 2019. 115 p. graf, ilus, tab.
Thesis in Portuguese | LILACS, BBO | ID: biblio-1051141

ABSTRACT

A redução da reatividade vascular à fenilefrina (PE) em aorta de ratas espontaneamente hipertensas (SHR) ao final da prenhez é dependente de maior produção e/ou maior biodisponibilidade de óxido nítrico (NO), consequente do aumento da fosforilação da enzima óxido nítrico sintase endotelial (eNOS) via PI3K/Akt. A glicosilação do tipo N-acetil-glucosamina (O-GlcNAc) é uma modificação pós-traducional que compete com a fosforilação pelos mesmos sítios de ligação nas proteínas. A O-GlcNAcilação da eNOS em serina1177 leva a redução da sua atividade enquanto a fosforilação leva a sua ativação. Além destes mecanismos, a interação da eNOS com outras proteínas é capaz de regular positiva ou negativamente a sua atividade. O objetivo deste trabalho foi analisar possíveis alterações nos mecanismos de modificação pós-traducional que controlam a ativação da eNOS os quais poderiam contribuir para maior ativação e maior biodisponibilidade de NO observada em artérias de ratas prenhes. Foram avaliados o conteúdo proteico O-GlcNAc e também expressão das enzimas que participam desta modificação, O-GlcNAc transferase (OGT) e O-GlcNAcase (OGA) por Western Blotting e a atividade da OGA por ensaio bioquímico em aorta e em artéria mesentérica (2º ou 3º ramo) de ratas não prenhes (NP) e prenhes (P), normotensas (Wistar) e SHR. Ensaios de Western Blotting foram realizados também para análise da expressão das seguintes proteínas: Cav-1, p-Cav-1, CaM e Hsp90. Realizamos a contagem do número de cavéolas endoteliais da aorta e da artéria mesentérica na presença ou ausência da metil-ß-ciclodextrina (dextrina, 10 mmol/L) por microscopia eletrônica. Em estudos funcionais, avaliamos a participação da enzima OGA, pela inibição com PugNAc (100 µmol/L) e das cavéolas, utilizando um desorganizador de cavéolas, a dextrina (10 ou 20 mmol/L), na menor reatividade vascular à PE observada em aortas de ratas P. Observamos que o conteúdo de proteínas O-GlcNAciladas estava diminuído em aorta e em leito mesentérico de ratas Wistar P e SHR P. Apesar da expressão da OGT e da OGA não estar alterada, a atividade da OGA foi aumentada em aorta e leito mesentérico de ratas Wistar P, mas, encontra-se diminuída em aorta e aumentada em leito mesentérico de SHP P. A incubação com PugNAc reverteu a reduzida reatividade à PE em aorta e artéria mesentérica de ratas Wistar P mas este efeito não foi observado em vasos SHR P, demonstrando que a OGA parece ter um papel importante na redução da O-GlcNAcilação de proteínas vasculares em Wistar P. Em vasos incubados com PugNAc, a remoção do endotélio ou a incubação com L-NAME, não alterou significativamente a reatividade à PE. Juntos estes resultados sugerem que a maior atividade da eNOS observada em vasos de Wistar P, fica prejudicada na presença do PugNAc, e depende da atividade da OGA. Como não houve alteração da resposta contrátil à PE em vasos de SHR P incubados com PugNAc, possivelmente um mecanismo diferente, envolvendo a menor atividade da OGT, ocorre nestas artérias para a redução da O-GlcNAcilação da eNOS. A desorganização das cavéolas por meio da dextrina causou aumento de contração à PE e redução de potência da ACh em aortas de Wistar NP e SHR NP, porém não houve alteração em aortas de ratas Wistar P e SHR P. A dextrina não alterou o número de cavéolas em artérias de Wistar P e SHR P quando comparado com ratas NP. SHR NP apresentam um reduzido número de cavéolas das aortas em relação a Wistar NP bem como expressão reduzida de Cav-1, p-Cav-1 e CaM. A prenhez não foi capaz de alterar a expressão da Cav-1, CaM e Hsp90 em aorta e leito mesentérico de ratas normotensas e hipertensas. Estes resultados sugerem que a prenhez não altera a expressão das proteínas Cav-1, CaM e Hsp90 e possivelmente a interação com a eNOS em aorta e artérias mesentéricas de ratas normotensas e hipertensas. Em conclusão, entre os mecanismos estudados de modificação pós-traducional da eNOS, a redução da O-GlcNAcilação da eNOS, por mecanismos que envolvem a atividade da OGA e possivelmente da OGT, favoreceria a fosforilação da eNOS e consequente maior biodisponibilidade de NO, contribuindo desta forma para modulação da resposta contrátil da PE nas artérias de ratas P(AU)


Reduction of vascular reactivity to phenylephrine (PE) in aorta of spontaneously hypertensive rats (SHR) at the end of pregnancy is dependent on higher production and/or higer bioavailability of nitric oxide (NO), as a consequence of increased endothelial nitric oxide synthase enzyme (eNOS) phosphorylation, by PI3K/Akt. Glycosylation with O-linked N-acetylglucosamine (O-GlcNAc) is a post-translational modification that competes with phosphorylation by the same binding sites in proteins. O-GlcNAcylation of eNOS on serine site leads to a reduction in its activity while eNOS phosphorylation leads to its activation. In addition to these mechanisms, the interaction of eNOS with other proteins is able to regulate positively or negatively its activity. The objective of this study was to analyze possible changes in the mechanisms of post-translational modification that control the eNOS activation, which could contribute to its the greater activation and greater bioavailability of NO observed in arteries of pregnant rats. The O-GlcNAc-protein content and also the enzymes expression that participate in this modification, O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA) was assessed by Western Blotting, and OGA activity were evaluated by biochemical assay in the aorta and in the artery mesenteric (2nd or 3rd branch) of non-pregnant (NP) and pregnant (P), normotensive rats (Wistar) and SHR. Western Blotting assays were also performed for expression analysis of the following proteins: Cav-1, p-Cav-1, CaM and Hsp90. We performed the counting of the number of endothelial caveolae in the aorta and the mesenteric artery in the presence or absence of methyl-ß-cyclodextrin (dextrin, 10 mmol/L) by electronic microscopy. In functional studies, we evaluated the participation of the OGA enzyme, by inhibition with PugNAc (100 µmol/L) and of the caveolae, using a caveolae disassembler, dextrin (10 or 20 mmol/L), in the reduced vascular reactivity observed in aortas or mesenteric arteries of P rats. We observed that the content of O-GlcNAcylated proteins was decreased in the aorta and in the mesenteric bed of Wistar P and SHR P rats. Although OGT and OGA expression is not altered, OGA activity was increased in the aorta and mesenteric bed of Wistar P rats but was decreased in the aorta and increased in the mesenteric bed of SHP P. Incubation with PugNAc reversed the reduced reactivity to PE in the aorta and mesenteric artery of Wistar P but this effect was not observed in SHR P arteries, demonstrating that OGA appears to play an important role in reducing O-GlcNAcylation of vascular proteins in Wistar P. In arteries incubated with PugNAc, endothelial removal or incubation with L-NAME did not significantly alter reactivity to PE. Together, these results suggest that the greater eNOS activity observed in Wistar P vessels was impaired in the presence of PugNAc, and it depends on OGA activity. As there was no change in the contractile response to PE in SHR P arteries incubated with PugNAc, possibly a different mechanism, involving the lower activity of OGT, occurs in these vessels for the reduction of O-GlcNAcylation of eNOS. Dextrin caused increased contraction of PE and decreased ACh potency in Wistar NP and SHR NP aortas, but there was no change in aortas of Wistar P and SHR P. Dextrin did not alter the number of cavelae in Wistar P and SHR P arteries compared to NP rats. SHR NP showed a lower number of caveolae than to NP Wistar as well reduced expression of Cav-1 and CaM. Pregnancy was not able to alter the expression of Cav-1, CaM and Hsp90 in the aorta and mesenteric bed of normotensive and hypertensive rats. These results suggest that pregnancy does not alter the expression of Cav-1, CaM and Hsp90 proteins and possibly interaction with eNOS in the aorta and mesenteric arteries of normotensive and hypertensive rats. In conclusion, among the studied mechanisms of post-translational modification of eNOS, the reduction of O-GlcNAcylation of eNOS, by mechanisms that involve OGA activity and possibly OGT, would favor eNOS phosphorylation and consequent greater NO bioavailability, contributing in this way for modulation of the contractile response to PE in the arteries of P rats(AU)


Subject(s)
Animals , Female , Pregnancy , Aorta , Nitric Oxide Synthase , Hypertension , Glycosylation , Calmodulin , Rats, Wistar , HSP90 Heat-Shock Proteins , Caveolin 1 , Mesenteric Arteries
7.
Acta Physiologica Sinica ; (6): 792-798, 2019.
Article in Chinese | WPRIM | ID: wpr-777131

ABSTRACT

Aberrant oxidative metabolism in cells is one of the hallmarks of cancer. Overproduction of reactive species promotes carcinogenesis by inducing genetic mutations and activating oncogenic pathways, and thus, antioxidant therapy is considered as an important strategy for cancer prevention and treatment. Caveolin-1 (Cav-1), a constituent protein of caveolae, is involved in not only the formation of the caveolae, vesicular transport, maintaining cholesterol homeostasis directly, but also many cellular physiological and pathological processes including growth, regulation of mitochondrial antioxidant level, apoptosis and carcinomas by interacting with a lot of signaling molecules through caveolin scaffolding domain. Cav-1 has also been shown to mediate tumor genesis and progression through oxidative stress modulation, while Cav-1-targeted treatment could scavenge the reactive species. Intracellular reactive species could modulate the expression, degradation, post-translational modifications and membrane trafficking of Cav-1. More importantly, emerging evidence has indicated that multiple antioxidants could exert antitumor activities in cancer cells by modulating the signaling of Cav-1. This paper reviewed the research progresses on the roles of Cav-1 and oxidative stress in tumorigenesis and development, and would provide new insights on designing strategies for cancer prevention or treatment.


Subject(s)
Antioxidants , Apoptosis , Carcinogenesis , Carcinoma , Pathology , Caveolin 1 , Humans , Mitochondria , Neoplasms , Pathology , Oxidative Stress , Signal Transduction
8.
Experimental Neurobiology ; : 377-386, 2018.
Article in English | WPRIM | ID: wpr-717414

ABSTRACT

Ubiquitin C-terminal hydrolase L1 (UCH-L1) is a deubiquitinating enzyme that is highly expressed in neurons, and gathering evidence indicates that UCH-L1 may play pathogenic roles in many neurodegenerative disorders such as Alzheimer's disease and Parkinson's disease (PD). Additionally, lipid rafts have attracted interest in neurodegeneration as playing a common role in many neurodegenerative diseases. In the present study, we demonstrated that UCH-L1 associates with lipid rafts as with other PD-associated gene products. In addition, UCH-L1 regulates lipid raft-dependent endocytosis and it is not dependent on the expression and degradation of caveolin-1 or flotillin-1. Finally, UCH-L1 regulates cell-to-cell transmission of α-synuclein. This study provides evidence that many PD-associated gene products share common signaling pathways to explain the pathogenesis of PD.


Subject(s)
alpha-Synuclein , Alzheimer Disease , Caveolin 1 , Endocytosis , Neurodegenerative Diseases , Neurons , Parkinson Disease , Prion Diseases , Ubiquitin Thiolesterase , Ubiquitin
9.
Odontoestomatol ; 19(30): 92-98, dic 2017.
Article in English, Spanish | LILACS, BNUY, BNUY-Odon | ID: biblio-876323

ABSTRACT

Las caveolas son invaginaciones de la membrana plasmática conformada por proteínas denominadas caveolinas. De las tres isoformas de caveolina la más estudiada en el transcurso de los años es caveolina-1 (cav-1), quien juega un papel importante en la señalización celular y su gen CAV-1 corresponde a la familia de genes supresores tumorales. Debido a su rol dependiente del contexto en el cual se encuentra, la función y participación de cav-1 a nivel tumoral es compleja y aún permanece controvertida. Cav-1 interactúa con una serie de moléculas-receptores que regulan inicialmente los pasos de la transformación celular a la malignidad, participando a su vez en el ciclo celular, la angiogénesis, la remodelación de la matriz extracelular, la proliferación celular, entre otras. El estudio de esta molécula adquiere importancia en función a su utilidad como biomarcador asociado a diagnóstico, pronóstico y posible blanco terapéutico en procesos patológicos.


Caveolae are plasma membrane invaginations formed by proteins called caveolins. Of the three caveolin isoforms, the most studied one through years has been caveolin-1 (cav-1), which has an important role in cell signaling, and its gene, CAV-1, is part of the family of tumor suppressor genes. As its role depends on the context, the participation and function of cav-1 in tumors is complex and remains controversial. Cav-1 interacts with a series of receptors and molecules that regulate the initial steps of cellular transformation to malignity. It also participates in the cell cycle, angiogenesis, extracellular matrix remodeling, cell proliferation, among other processes. The study of this molecule is important due to its function as a biomarker associated to the diagnosis, prognosis and therapeutic target in pathological processes.


Subject(s)
Caveolin 1 , Genes, Tumor Suppressor
10.
Acta cir. bras ; 32(5): 359-368, May 2017. tab, graf
Article in English | LILACS | ID: biblio-837709

ABSTRACT

Abstract Purpose: To evaluate the changes of caveolin-1 in lung fibroblasts in newborn Wistar rats when exposed to hyperoxic conditions, as well as lung fibroblasts cell cycle. Methods: One hundred newborn Wistar rats were randomly divided (50 rats/group) into experimental and control groups, exposed to hyperoxic conditions or normal air, respectively. The fraction of inspired oxygen (FiO2) in the experimental group was 90%, whereas this value was 21% in the control group. Lung fibroblasts were collected on days 3, 7, and 14 of the experiment. Caveolin-1 expression dynamics in lung fibroblasts was assayed in each group by immunofluorescence and Western blot analyses. Flow cytometry (FCM) was used to assess the proportions of lung fibroblasts at different stages of the cell cycle. Results: On day 3, no significant difference in caveolin-1 expression was observed between the hyperoxic and control groups; however, on days 7 and 14, caveolin-1 expression was significantly lower in the hyperoxic group than in the control (P<0.05). No apparent differences were observed in caveolin-1 expression in the control group at the different time points. Using FCM analysis, we showed that the proportion of lung fibroblasts in G0/G1 phase in the hyperoxic group decreased compared to that of the control group on day 7, while the proportion of S-phase cells increased (P<0.05). These differences were more significant when the groups were compared on day 14 (P<0.01). Conclusion: After seven days the exposure to hyperoxic conditions, lung fibroblasts proliferated and caveolin-1 expression decreased.


Subject(s)
Animals , Female , Cell Proliferation , Caveolin 1/metabolism , Fibroblasts/metabolism , Lung/metabolism , Lung Diseases/metabolism , Oxygen/pharmacology , Random Allocation , Cell Cycle , Cells, Cultured , Chronic Disease , Rats, Wistar , Hyperoxia , Models, Animal , Caveolin 1/pharmacology , Fibroblasts/cytology , Fibroblasts/drug effects , Lung/cytology , Lung/drug effects , Lung Diseases/classification , Lung Diseases/chemically induced , Animals, Newborn
11.
Acta Physiologica Sinica ; (6): 252-260, 2017.
Article in Chinese | WPRIM | ID: wpr-348277

ABSTRACT

To study the correlation between the spatial cognitive impairment and different subtypes of estrogen receptor α (ERα) of hippocampus in diabetic mice, we used alloxan (intraperitoneal injection) to induce type 1 diabetes in male Kunming mice and compared the spatial cognitive ability of the model mice with that of control mice through Morris water maze test. Meanwhile, using Western blot, we detected the protein expressions of ER-α36, ER-α66, caveolin-1, PKCα, cAMP-response element binding protein 2 (CREB2), and synaptophysin (Syn) in the hippocampus of the mice. The results showed that on the 3rd and 5th days of training, the ability of spatial learning and memory in the diabetic mice was significantly inferior to that of the control mice (P < 0.05). In the diabetic mice, the protein expressions of caveolin-1 and PKCα were decreased (P < 0.05), but ER-α66 expression was unaffected, while ER-α36 and CREB2 expressions were significantly increased (P < 0.05) compared with those of the control mice. The results suggest that abnormal expression of ER-α36 and related signal molecules may be important factors for diabetes-induced spatial cognitive impairment.


Subject(s)
Animals , Caveolin 1 , Metabolism , Cognitive Dysfunction , Cyclic AMP Response Element-Binding Protein , Metabolism , Diabetes Mellitus, Experimental , Estrogen Receptor alpha , Metabolism , Hippocampus , Metabolism , Male , Maze Learning , Memory , Mice , Protein Kinase C-alpha , Metabolism , Synaptophysin , Metabolism
12.
Chonnam Medical Journal ; : 28-36, 2017.
Article in English | WPRIM | ID: wpr-788363

ABSTRACT

Caveolin-1 (Cav-1) is a trans-membrane protein that is a major component of the caveolae structure on the plasma membrane. Cav-1 is involved in the regulation of various cellular processes, including cell growth, differentiation, endocytosis, and in particular it has been implied in cellular senescence. Here we review current knowledge about Cav-1 in cellular signaling and discuss the role of Cav-1 in aging-related diseases.


Subject(s)
Caveolae , Caveolin 1 , Cellular Senescence , Cell Membrane , Endocytosis
13.
Article in English | WPRIM | ID: wpr-151261

ABSTRACT

BACKGROUND/AIMS:: We demonstrated the role of caveolin-1 involved in high glucose (HG)-induced glomerular mesangial cells (GMCs) senescence. METHODS:: HG was used to stimulate GMCs. The telomere lengths were analyzed by Southern blot. β-Galactosidase staining was determined. The expressions of caveolin-1 and P53 proteins were determined by Western blot. RESULTS:: Treatment with high concentrations of glucose induced GMC senescence accompanied by shortened telomere length and increase of β-galactosidase staining as well as P53 protein, which was abrogated after application of caveolin-1-siRNA. CONCLUSIONS:: This study proved that HG induced cell senescence in GMCs. The caveolin-1 is involved in HG-induced mesangial cell senescence, and blocking caveolin-1 significantly reduced cell senescence. The effect of caveolin-1 is mediated by P53 pathway.


Subject(s)
Aging , Blotting, Southern , Blotting, Western , Caveolin 1 , Cellular Senescence , Glucose , Humans , Mesangial Cells , Telomere
14.
National Journal of Andrology ; (12): 11-20, 2017.
Article in Chinese | WPRIM | ID: wpr-812817

ABSTRACT

Objective@#To investigate whether androgens can regulate the expression of eNOS in rat corpus cavernosum through AKT3, PIK3CA, CALM, and CAV1 and influence erectile function.@*METHODS@#Thirty-six 8-week-old male SD rats were randomly divided into groups A (4-week control), B (6-week control), C (4-week castration), D (6-week castration), E (4-week castration + testosterone replacement), and F (6-week castration + testosterone replacement). Both the testis and epididymis were removed from the rats in groups C, D, E and F, and on the second day after surgery, the animals of groups E and F were subcutaneously injected with testosterone propionate at 3 mg per kg of the body weight qd alt while all the others with isodose oil instead. At 4 weeks (for groups A, C and E) and 6 weeks (for groups B, D and F) after treatment, we detected the maximum intracavernous pressure (ICPmax), the mean carotid arterial pressure (MAP) and their ratio (ICPmax/MAP), measured the level of serum testosterone (T), and determined the expressions of eNOS, P-eNOS, AKT3, PIK3CA, CALM and CAV1 in the corpus cavernosum by Western blot and immunohistochemistry.@*RESULTS@#No statistically significant differences were observed in the body weight and MAP among different groups. The serum T level and ICPmax/MAP were remarkably lower in groups C and D than in the other four groups (P<0.01) as well as in groups E and F than in A and B (P<0.05) but exhibited no significant differences either between E and F or between A and B. Immunohistochemistry showed that eNOS and P-eNOS were mainly expressed in the vascular endothelial cell membrane and cavernous vascular lumen, while AKT3, PIK3CA, CALM and CAV1 chiefly in the vascular endothelial cell cytoplasm and membrane, with a few in the smooth muscle cells. Western blot analysis manifested that the expressions of eNOS, P-eNOS, AKT3, PIK3CA, CALM and CAV1 were markedly lower in groups C and D than in A, B, E and F (P<0.01) as well as in D than in C (P<0.05) but those in groups E and F did not showed any significant difference from those in A and B, nor E from F or A from B.@*CONCLUSIONS@#Androgens can improve erectile function by upregulating the expressions of AKT3, PIK3CA, CALM and CAV1 protein molecules and activating eNOS after its phosphorylation, though the exact molecular mechanisms are yet to be further studied.


Subject(s)
Animals , Blood Pressure , Blotting, Western , Caveolin 1 , Metabolism , Class I Phosphatidylinositol 3-Kinases , Metabolism , Erectile Dysfunction , Hormone Replacement Therapy , Male , Monomeric Clathrin Assembly Proteins , Metabolism , Myocytes, Smooth Muscle , Nitric Oxide Synthase Type III , Metabolism , Orchiectomy , Penile Erection , Physiology , Penis , Metabolism , Proto-Oncogene Proteins c-akt , Metabolism , Random Allocation , Rats , Rats, Sprague-Dawley , Testosterone Propionate
15.
Chonnam Medical Journal ; : 28-36, 2017.
Article in English | WPRIM | ID: wpr-67657

ABSTRACT

Caveolin-1 (Cav-1) is a trans-membrane protein that is a major component of the caveolae structure on the plasma membrane. Cav-1 is involved in the regulation of various cellular processes, including cell growth, differentiation, endocytosis, and in particular it has been implied in cellular senescence. Here we review current knowledge about Cav-1 in cellular signaling and discuss the role of Cav-1 in aging-related diseases.


Subject(s)
Caveolae , Caveolin 1 , Cellular Senescence , Cell Membrane , Endocytosis
16.
Araçatuba; s.n; 2016. 107 p. ilus, graf, tab.
Thesis in Portuguese | LILACS, BBO | ID: biblio-881587

ABSTRACT

O TERPY promove efeito hipotensor de maior magnitude em ratos hipertensos (SHR e 2R-1C) do que em ratos normotensos (Wistar e 2R). Foi demonstrado anteriormente que o endotélio prejudica o efeito vasodilatador do TERPY em aorta de Wistar. No entanto, observamos que o endotélio melhora os efeitos vasodilatadores do TERPY em aorta de SHR. Vasos sanguíneos de menor calibre, tais como as artérias de resistência, estão associadas ao controle da resitência vascular periférica e da pressão arterial. Nossa hipótese é que o TERPY induz relaxamento nas artérias mesentéricas de resistência em SHR e que as células endoteliais modulam positivamente o efeito do TERPY nestes vasos sanguíneos. Portanto, o nosso objetivo foi avaliar o efeito vasodilatador do TERPY em anéis com e sem endotélio de artéria mesentérica de ratos SHR, o seu mecanismo de relaxamento e a participação da NOS sobre esse efeito. e a Nossos resultados mostraram que o TERPY induziu um efeito vasodilatador dependente da concentração em anéis de artérias mesentéricas (2º e 3º ramos) de SHR e de ratos Wistar. A potência do TERPY foi maior em anéis intactos do que em anéis sem endotélio em artérias mesentéricas de SHR, mas em Wistar o endotélio prejudicou o efeito do TERPY. Nas artérias mesentéricas sem endotélio de SHR, o efeito do TERPY é dependente da atividade da guanilato ciclase solúvel e de canais para potássio. Nas artérias mesentéricas intactas de SHR, o efeito de TERPY depende da atividade de eNOS, mas não é dependente das atividades de nNOS, iNOS ou da via da ciclooxigenase. O TERPY promove a fosforilação da eNOS nos resíduos de serina1177 e aumenta a concentração de óxido nítrico em células endoteliais isoladas de artérias mesentéricas de SHR. Nossos resultados mostraram que a guanilato ciclase solúvel, os canais para potássio e a eNOS estão envolvidos no efeito vasodilatador estimulado pelo TERPY nas artérias de resistência mesentérica de SHR. Numa segunda parte deste estudo, avaliamos o mecanismo de ação de TERPY e seu efeito sobre a atividade da eNOS em células endoteliais. As células HUVEC, WT-HEK e HEK-eNOS foram tratadas com TERPY em diferentes tempos (0 a 60 minutos). Foram analisados por Western blotting o efeito do TERPY sobre a fosforilação de eNOS, monômero e dímero da eNOS e sobre monômero e oligômero de caveolina-1. Também foi avaliado o efeito do TERPY na interação eNOS/Cav-1 através de co-imunoprecipitação. As alterações induzidas pelo TERPY sobre as concentrações de espécies reativas de oxigênio e peroxinitrito em células endoteliais foram medidas usando sonda DHE e biossensor 7-CBA, respectivamente. A concentração de óxido nítrico (NO) foi avaliada por sonda DAF-FM e sensor Cooper. O TERPY promoveu desacoplamento e disfunção da eNOS, dependente de BH4. A desestabilização dos oligômeros da caveolina-1 foi induzida pelo TERPY. Consequentemente, o TERPY reduziu a interação eNOS/Cav-1 e promoveu ativação da eNOS. Nossos resultados mostraram que a atividade da eNOS pode ser regulada de duas maneiras diferentes pelo TERPY. O TERPY promove desacoplamento e fosforilação da eNOS, promovendo uma estratégia diferente para a regulação da atividade desta enzima. As moléculas químicas ou biológicas como o TERPY que regulam a atividade da eNOS e aumentam a produção e a biodisponibilidade de NO teriam ações terapêuticas importantes para o tratamento de doenças vasculares associadas a hipertensão(AU)


TERPY promotes a hypotensive effect with greater magnitude in hypertensive rats (SHR and 2K-1C) than in normotensive rats (Wistar and 2K). Previously, it was demonstrated that endothelium impairs vasodilatory effect of TERPY in Wistar aorta. However, we observed that endothelium improves the vasodilatory effect of TERPY in SHR aorta. Smaller blood vessels, such as mesenteric arteries, are associated with the control of peripheral vascular resistance and blood pressure. We hypothesized that TERPY induces relaxation on mesenteric resistance arteries in SHR and endothelial cells modulate positively the TERPY's effect on these blood vessels. Therefore, our goal was to evaluate the vasodilator effect of TERPY in rings with and without endothelium of mesenteric arteries in SHR, the mechanism of relaxation and the participation of NOS on this effect. Our results show TERPY induced a concentration-dependent vasodilator effect in mesenteric arteries (2nd and 3 rd branches) rings from SHR and Wistar. The potency of TERPY was higher in intact than in denuded rings from SHR, but in Wistar, endothelium impair the TERPY's effect. In denuded mesenteric arteries from SHR, the relaxation effect induced by TERPY is dependent of soluble guanylate cyclase and activation of potassium channel. However, in intact mesenteric arteries from SHR, TERPY´s effect is modulated by eNOS activity, but it is not dependent of nNOS, iNOS or cyclooxygenase pathway activities. TERPY promotes eNOS3 Ser1177 phosphorylation and increases nitric oxide concentration in isolated endothelial cells of mesenteric arteries from SHR. Together, our results showed that soluble guanylate cyclase, potassium channels, and eNOS are involved in the vasodilator effect of TERPY in mesenteric resistance arteries from SHR. In a second part of this study, we aimed to evaluate the mechanism of action of TERPY and its effect on eNOS activity in endothelial cells. HUVEC, WT-HEK and HEK-eNOS cells were treated with TERPY at different times (0 to 60 minutes). Were analyzed by western blotting the TERPY`s effect on eNOS monomer, dimer and phosphorylation and on monomer and oligomer of caveolin-1. It was also evaluated the effect of TERPY in the interaction between eNOS/Cav-1 through co-immunoprecipitation. Alterations induced by TERPY on reactive oxygen species and peroxynitrite concentrations in endothelial cells were measured by using DHE probe and biosensor 7-CBA, respectively. Nitric oxide fluorescence was assessed by DAF-FM probe and Cooper sensor. TERPY promoted eNOS uncoupling and eNOS dysfunction, which is BH4-dependent. Caveolin-1 oligomers destabilization was induced by TERPY. Consequently, TERPY reduced eNOS/Cav-1 interaction and promoted eNOS activation. Our results show that eNOS activity can be regulated in two different ways by TERPY, leading to eNOS uncoupling and leading to eNOS phosphorylation, promoting a strategy for eNOS regulation. Chemical or biological molecules as TERPY that regulates eNOS activity and increase NO production and bioavailability are potential therapeutic drugs for the treatment of vascular diseases associated with hypertension(AU)


Subject(s)
Animals , Rats , Nitric Oxide , Nitric Oxide Synthase Type III , Rats, Inbred SHR , Vasodilation , Caveolin 1 , Endothelium, Vascular , Hypertension , Mesenteric Arteries , Vascular Diseases
17.
Journal of Stroke ; : 12-20, 2016.
Article in English | WPRIM | ID: wpr-135899

ABSTRACT

Moyamoya disease (MMD) is a unique cerebrovascular disease characterized by the progressive stenosis of large intracranial arteries and a hazy network of basal collaterals called moyamoya vessels. Because the etiology of MMD is unknown, its diagnosis is based on characteristic angiographic findings. Re-vascularization techniques (e.g., bypass surgery) are used to restore perfusion, and are the primary treatment for MMD. There is no specific treatment to prevent MMD progression. This review summarizes the recent advances in MMD pathophysiology, including the genetic and circulating factors related to disease development. Genetic and environmental factors may play important roles in the development of the vascular stenosis and aberrant angiogenesis in complex ways. These factors include the related changes in circulating endothelial/smooth muscle progenitor cells, cytokines related to vascular remodeling and angiogenesis, and endothelium, such as caveolin which is a plasma membrane protein. With a better understanding of MMD pathophysiology, nonsurgical approaches targeting MMD pathogenesis may be available to stop or slow the progression of this disease. The possible strategies include targeting growth factors, retinoic acid, caveolin-1, and stem cells.


Subject(s)
Arteries , Caveolin 1 , Cell Membrane , Constriction, Pathologic , Cytokines , Diagnosis , Endothelium , Intercellular Signaling Peptides and Proteins , Moyamoya Disease , Perfusion , Stem Cells , Tretinoin
18.
Journal of Stroke ; : 12-20, 2016.
Article in English | WPRIM | ID: wpr-135894

ABSTRACT

Moyamoya disease (MMD) is a unique cerebrovascular disease characterized by the progressive stenosis of large intracranial arteries and a hazy network of basal collaterals called moyamoya vessels. Because the etiology of MMD is unknown, its diagnosis is based on characteristic angiographic findings. Re-vascularization techniques (e.g., bypass surgery) are used to restore perfusion, and are the primary treatment for MMD. There is no specific treatment to prevent MMD progression. This review summarizes the recent advances in MMD pathophysiology, including the genetic and circulating factors related to disease development. Genetic and environmental factors may play important roles in the development of the vascular stenosis and aberrant angiogenesis in complex ways. These factors include the related changes in circulating endothelial/smooth muscle progenitor cells, cytokines related to vascular remodeling and angiogenesis, and endothelium, such as caveolin which is a plasma membrane protein. With a better understanding of MMD pathophysiology, nonsurgical approaches targeting MMD pathogenesis may be available to stop or slow the progression of this disease. The possible strategies include targeting growth factors, retinoic acid, caveolin-1, and stem cells.


Subject(s)
Arteries , Caveolin 1 , Cell Membrane , Constriction, Pathologic , Cytokines , Diagnosis , Endothelium , Intercellular Signaling Peptides and Proteins , Moyamoya Disease , Perfusion , Stem Cells , Tretinoin
19.
Journal of Breast Cancer ; : 372-384, 2016.
Article in English | WPRIM | ID: wpr-28542

ABSTRACT

PURPOSE: Lipid rafts are cholesterol enriched microdomains that colocalize signaling pathways involved in cell proliferation, metastasis, and angiogenesis. We examined the effect of methyl-β-cyclodextrin (MβCD)-mediated cholesterol extraction on the proliferation, adhesion, invasion, and angiogenesis of triple negative breast cancer (TNBC) cells. METHODS: We measured cholesterol and estimated cell toxicity. Detergent resistant membrane (DRM) and non-DRM fractions were separated using the OptiPrep gradient method. Cell cycles stages were analyzed by flow cytometry, apoptosis was assessed using the TdT-mediated dUTP nick end-labeling assay, and metastasis was determined using a Matrigel invasion assay. Neo-vessel pattern and levels of angiogenic modulators were determined using an in vitro angiogenesis assay and an angiogenesis array, respectively. RESULTS: The present study found that the cholesterol-depleting agent MβCD, efficiently depleted membrane cholesterol and caused concentration dependent (0.1–0.5 mM) cytotoxicity compared to nystatin and filipin III in TNBC cell lines, MDA-MB 231 and MDA-MB 468. A reduced proportion of caveolin-1 found in DRM fractions indicated a cholesterol extraction-induced disruption of lipid raft integrity. MβCD inhibited 52% of MDA-MB 231 cell adhesion on fibronectin and 56% of MDA-MB 468 cell adhesion on vitronectin, while invasiveness of these cells was decreased by 48% and 52% respectively, following MβCD treatment (48 hours). MβCD also caused cell cycle arrest at the G2M phase and apoptosis in MDA-MB 231 cells (25% and 58% cells, respectively) and in MDA-MB 468 cells (30% and 38% cells, respectively). We found that MβCD treated cells caused a 52% and 58% depletion of neovessel formation in both MDA-MB 231 and MDA-MB 468 cell lines, respectively. This study also demonstrated that MβCD treatment caused a respective 2.6- and 2.5-fold depletion of tyrosine protein kinase receptor (TEK) receptor tyrosine kinase levels in both TNBC cell lines. CONCLUSION: MβCD-induced cholesterol removal enhances alterations in lipid raft integrity, which reduces TNBC cell survival.


Subject(s)
Apoptosis , Caveolin 1 , Cell Adhesion , Cell Cycle , Cell Cycle Checkpoints , Cell Line , Cell Proliferation , Cell Survival , Cholesterol , Detergents , Fibronectins , Filipin , Flow Cytometry , In Vitro Techniques , Membrane Microdomains , Membranes , Methods , Neoplasm Metastasis , Nystatin , Protein-Tyrosine Kinases , Triple Negative Breast Neoplasms , Vitronectin
20.
Article in English | WPRIM | ID: wpr-26789

ABSTRACT

PURPOSE: Caveolin-1 (CAV-1) expression is more associated with basal-like cancers than estrogen receptor- or ErbB-2-expressing breast cancers. However, the biological relevance of different levels of CAV-1 expression according to subtype in the epithelial compartment of breast cancer remains unclear. MATERIALS AND METHODS: We investigated whether CAV-1 functions as a tumor suppressor and/or modulator of the cytotoxic activity of docetaxel (DTX) in subtypes of breast cancer using in vitro and xenograft models. RESULTS: The levels of CAV-1 expression were closely associated with DTX sensitivity in triple-negative breast cancer cells. In addition, CAV-1 significantly inhibited cell proliferation and modulated DTX-induced apoptosis through cell cycle arrest in the G2/M phase. The mechanisms underlying DTX-induced apoptosis differed in breast cancers according to the levels of CAV-1 expression. DTX robustly enhanced Bcl-2 inactivation by CAV-1 in MDA-MB-231 cells, while p53-mediated cell cycle arrest by DTX was more pronounced in CAV-1-low but p53-functional MCF-7 cells. In parallel with the data from breast cancer cell lines, CAV-1-transfected MCF-7 cells showed higher efficacy of DTX treatment in a xenograft model. CONCLUSION: We clearly demonstrated cooperative effects between CAV-1 and DTX in mediating apoptosis, suggesting that the levels of CAV-1 expression might be an important indicator for DTX use in breast cancer.


Subject(s)
Apoptosis , Breast Neoplasms , Breast , Caveolin 1 , Cell Cycle Checkpoints , Cell Death , Cell Line , Cell Proliferation , Estrogens , Heterografts , MCF-7 Cells , Negotiating , Triple Negative Breast Neoplasms
SELECTION OF CITATIONS
SEARCH DETAIL