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1.
Int J Mol Sci ; 25(2)2024 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-38255846

RESUMO

PC12 cells, which are derived from rat adrenal pheochromocytoma cells, are widely used for the study of neuronal differentiation. NGF induces neuronal differentiation in PC12 cells by activating intracellular pathways via the TrkA receptor, which results in elongated neurites and neuron-like characteristics. Moreover, the differentiation requires both the ERK1/2 and p38 MAPK pathways. In addition to NGF, BMPs can also induce neuronal differentiation in PC12 cells. BMPs are part of the TGF-ß cytokine superfamily and activate signaling pathways such as p38 MAPK and Smad. However, the brief lifespan of NGF and BMPs may limit their effectiveness in living organisms. Although PC12 cells are used to study the effects of various physical stimuli on neuronal differentiation, the development of new methods and an understanding of the molecular mechanisms are ongoing. In this comprehensive review, we discuss the induction of neuronal differentiation in PC12 cells without relying on NGF, which is already established for electrical, electromagnetic, and thermal stimulation but poses a challenge for mechanical, ultrasound, and light stimulation. Furthermore, the mechanisms underlying neuronal differentiation induced by physical stimuli remain largely unknown. Elucidating these mechanisms holds promise for developing new methods for neural regeneration and advancing neuroregenerative medical technologies using neural stem cells.


Assuntos
Neoplasias das Glândulas Suprarrenais , Animais , Ratos , Células PC12 , Diferenciação Celular , Estimulação Física , Proteínas Quinases p38 Ativadas por Mitógeno
2.
Int J Mol Sci ; 23(24)2022 Dec 09.
Artigo em Inglês | MEDLINE | ID: mdl-36555248

RESUMO

This study evaluated the mechanism of temperature-controlled repeated thermal stimulation (TRTS)-mediated neuronal differentiation. We assessed the effect of SP600125, a c-Jun N-terminal kinase (JNK) inhibitor, on neuronal differentiation of rat PC12-P1F1 cells, which can differentiate into neuron-like cells by exposure to TRTS or neurotrophic factors, including bone morphogenetic protein (BMP) 4. We evaluated neuritogenesis by incubating the cells under conditions of TRTS and/or SP600125. Cotreatment with SP600125 significantly enhanced TRTS-mediated neuritogenesis, whereas that with other selective mitogen-activated protein kinase (MAPK) inhibitors did not-e.g., extracellular signal-regulated kinase (ERK)1/2 inhibitor U0126, and p38 MAPK inhibitor SB203580. We tried to clarify the mechanism of SP600125 action by testing the effect of U0126 and the BMP receptor inhibitor LDN193189 on the SP600125-mediated enhancement of intracellular signaling. SP600125-enhanced TRTS-induced neuritogenesis was significantly inhibited by U0126 or LDN193189. Gene expression analysis revealed that TRTS significantly increased ß3-Tubulin, MKK3, and Smad7 gene expressions. Additionally, Smad6 and Smad7 gene expressions were substantially attenuated through SP600125 co-treatment during TRTS. Therefore, SP600125 may partly enhance TRTS-induced neuritogenesis by attenuating the negative feedback loop of BMP signaling. Further investigation of the mechanisms underlying the effect of SP600125 during TRTS-mediated neuritogenesis may contribute to the future development of regenerative neuromedicine.


Assuntos
Butadienos , Crescimento Neuronal , Animais , Ratos , Butadienos/farmacologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Células PC12 , Temperatura
3.
Int J Mol Sci ; 21(21)2020 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-33171774

RESUMO

Neuritogenesis is the process underling nervous system regeneration; however, optimal extracellular signals that can promote neuronal regenerative activities require further investigation. Previously, we developed a novel method for inducing neuronal differentiation in rat PC12 cells using temperature-controlled repeated thermal stimulation (TRTS) with a heating plate. Based on neurogenic sensitivity to TRTS, PC12 cells were classified as either hyper- or hyposensitive. In this study, we aimed to investigate the mechanism of hyposensitivity by establishing two PC12-derived subclones according to TRTS sensitivity during differentiation: PC12-P1F1, a hypersensitive subclone, and PC12-P1D10, a hyposensitive subclone. To characterize these subclones, cell size and neuritogenesis were evaluated in subclones treated with nerve growth factor (NGF), bone morphogenetic protein (BMP), or various TRTS. No significant differences in cell size were observed among the parental cells and subclones. BMP4- or TRTS-induced neuritogenesis was increased in PC12-P1F1 cells compared to that in the parental cells, while no neuritogenesis was observed in PC12-P1D10 cells. In contrast, NGF-induced neuritogenesis was observed in all three cell lines. Furthermore, a BMP inhibitor, LDN-193189, considerably inhibited TRTS-induced neuritogenesis. These results suggest that the BMP pathway might be required for TRTS-induced neuritogenesis, demonstrating the useful aspects of these novel subclones for TRTS research.


Assuntos
Regeneração Nervosa/fisiologia , Células PC12/metabolismo , Sensação Térmica/fisiologia , Animais , Diferenciação Celular/fisiologia , Neuritos/metabolismo , Neurogênese/fisiologia , Neurônios/metabolismo , Células PC12/fisiologia , Ratos , Temperatura
4.
Tohoku J Exp Med ; 248(3): 181-192, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31353327

RESUMO

Beta3-adrenergic receptor (ADRB3) is a mediator of catecholamine-stimulated lipolysis in humans. The Trp64Arg polymorphism with T/C transition in the ADRB3 gene has been considered to reduce lipolysis and metabolic expenditure. Here, we investigated the hitherto unknown role of the Trp64Arg substitution on food preference among healthy young adults (mean age, 24.3; n = 53, including 25 men). Preference toward four food types (bitter, sour, salty, or sweet) and greasy (high-fat) foods was examined using a self-reported questionnaire. There was no noticeable sex-difference in food preference. Incidentally, only among female subjects, the genotype frequencies of the Trp64Arg polymorphism were in accordance with the Hardy-Weinberg equilibrium. Consequently, female subjects were divided into two groups for further analyses: 18 subjects with TT genotype (Trp64Trp) (wild-type group) and 10 subjects with TC genotype (Trp64Arg) (heterozygous group). No significant difference was observed in preference for four food types between the groups. However, when sweet foods were divided into high-fat and low-fat subgroups, food preference for high-fat sweet foods in heterozygous group was significantly higher than that in wild-type group. Moreover, when subjects were divided into two classes based on preference for greasy foods (like, n = 16 or dislike, n = 12), the preference degree in heterozygous group who liked high-fat foods (n = 5) was significantly higher than that in wild-type group (n = 11), suggesting that the Trp64Arg substitution might genetically enhance high-fat food preference. Thus, understanding the relationship between ADRB3 Trp64Arg substitution and fat preference will be valuable for obesity prevention.


Assuntos
Gorduras na Dieta/administração & dosagem , Preferências Alimentares , Estudos de Associação Genética , Polimorfismo de Nucleotídeo Único/genética , Receptores Adrenérgicos beta 3/genética , Adulto , Índice de Massa Corporal , Feminino , Frequência do Gene , Humanos , Japão , Masculino , Inquéritos e Questionários , Adulto Jovem
5.
Respir Res ; 19(1): 53, 2018 04 02.
Artigo em Inglês | MEDLINE | ID: mdl-29606146

RESUMO

Background: Dopamine receptors comprise two subgroups, Gs protein-coupled "D1-like" receptors (D1, D5) and Gicoupled "D2-like" receptors (D2, D3, D4). In airways, both dopamine D1 and D2 receptors are expressed on airway smooth muscle and regulate airway smooth muscle force. However, functional expression of the dopamine D1 receptor has never been identified on airway epithelium. Activation of Gs-coupled receptors stimulate adenylyl cyclase leading to cyclic AMP (cAMP) production, which is known to induce mucus overproduction through the cAMP response element binding protein (CREB) in airway epithelial cells. We questioned whether the dopamine D1 receptor is expressed on airway epithelium, and whether it promotes CREB phosphorylation and MUC5AC expression. Methods: We evaluated the protein expression of the dopamine D1 receptor on native human airway epithelium and three sources of cultured human airway epithelial cells including primary cultured airway epithelial cells, the bronchial epithelial cell line (16HBE14o-), and the pulmonary mucoepidermoid carcinoma cell line (NCI-H292) using immunohistochemistry and immunoblotting. To characterize the stimulation of cAMP through the dopamine D1 receptor, 16HBE14o- cells and NCI-H292 cells were treated with dopamine or the dopamine D1 receptor agonists (SKF38393 or A68930) before cAMP measurements. The phosphorylation of CREB by A68930 in both 16HBE14o- and NCI-H292 cells was measured by immunoblot. The effect of dopamine or A68930 on the expression of MUC5AC mRNA and protein in NCI-H292 cells was evaluated by real-time PCR and immunofluorescence staining, respectively. Results: The dopamine D1 receptor protein was detected in native human airway epithelium and three sources of cultured human airway epithelial cells. Dopamine or the dopamine D1-like receptor agonists stimulated cAMP production in 16HBE14o- cells and NCI-H292 cells, which was reversed by the selective dopamine D1-like receptor antagonists (SCH23390 or SCH39166). A68930 significantly increased phosphorylation of CREB in both 16HBE14o- and NCI-H292 cells, which was attenuated by the inhibitors of PKA (H89) and MEK (U0126). Expression of MUC5AC mRNA and protein were also increased by either dopamine or A68930 in NCI-H292 cells. Conclusions: These results suggest that the activation of the dopamine D1 receptor on human airway epithelium could induce mucus overproduction, which could worsen airway obstructive symptoms.


Assuntos
Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Mucina-5AC/biossíntese , Receptores de Dopamina D1/biossíntese , Mucosa Respiratória/metabolismo , Linhagem Celular , Células Cultivadas , Agonistas de Dopamina/farmacologia , Expressão Gênica , Humanos , Mucina-5AC/genética , Fosforilação/efeitos dos fármacos , Fosforilação/fisiologia , Receptores de Dopamina D1/agonistas , Receptores de Dopamina D1/genética , Mucosa Respiratória/efeitos dos fármacos
6.
Tohoku J Exp Med ; 244(4): 249-261, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29576584

RESUMO

Oral fat sensitivity (OFS), the ability to taste fat, is associated with eating habits in humans, including daily fatty food consumption; namely, proper OFS is important for health. In addition, lifestyle factors, such as stress and sleep, may affect fat intake. However, correlations between OFS, stress management, and sleep are largely unknown. Thus, we investigated the associations of OFS with lifestyle factors regarding stress management and daytime sleepiness in healthy young adult men. We measured OFS, sensitivities for three prototypical tastes (sweet, salty, and sour), and BMI in 22 subjects (27.2 ± 6.0 years), who also completed a self-reported questionnaire on health, including lifestyle-related questions. The correlations of taste sensitivities with stress management and daytime sleepiness were analyzed. We thus found that the number of mentors for personal concerns (a question regarding stress management, social support) was positively associated with OFS (P = 0.041), but not the other taste sensitivities. In contrast, other lifestyle factors related to stress management (frequency of relaxation, frequency of eating when irritated or worried, and degree of satisfaction with one's sleep) and daytime sleepiness were not significantly associated with OFS, similar to that observed for other tested taste sensitivities. These results characterize the unique association of OFS with the degree of social support in healthy young men and provide novel insights into the nature of individual OFS differences. Thus, measuring the degree of social support may be helpful for the evaluation of OFS impairment, thereby contributing to the prevention of stress-induced overeating.


Assuntos
Adaptação Psicológica , Gorduras na Dieta , Apoio Social , Estresse Psicológico/terapia , Paladar , Administração Oral , Adulto , Índice de Massa Corporal , Comportamento Alimentar , Humanos , Estilo de Vida , Masculino , Sobrepeso/terapia , Sono , Sacarose/química , Inquéritos e Questionários , Adulto Jovem
7.
Tohoku J Exp Med ; 238(2): 93-103, 2016 02.
Artigo em Inglês | MEDLINE | ID: mdl-26797054

RESUMO

Oral fat sensitivity (OFS, the ability to detect fat) may be related to overeating-induced obesity. However, it is largely unknown whether OFS affects taste preference and eating habits. Therefore, we aimed to evaluate (1) the association between body mass index (BMI) and OFS and (2) the relationship of OFS with four types of taste preference (sweet, sour, salty, and bitter) and eating habits using serial concentrations of oleic acid (OA) homogenized in non-fat milk and a self-reported questionnaire. Participants were 25 healthy Japanese individuals (mean age: 27.0 ± 5.6 years), among whom the OA detection threshold was significantly associated with BMI. Participants were divided into two subgroups based on oral sensitivity to 2.8 mM OA: hypersensitive (able to detect 2.8 mM OA, n = 16) and hyposensitive (unable to detect 2.8 mM OA, n = 9). The degree of sweet taste preference of the hypersensitive group was significantly higher than that of the hyposensitive group. Furthermore, there was significantly higher degree of preference for high-fat sweet foods than low-fat sweet foods in the hypersensitive group. There was also a significant inverse correlation between the OA detection threshold and the degree of both spare eating and postprandial satiety. Thus, OFS is associated not only with BMI, but also with the preference for high-fat sweet foods and eating habits. The present study provides novel insights that measuring OFS may be useful for assessing the risk of obesity associated with overeating in countries, including Japan, where BMI is increasing in the population.


Assuntos
Índice de Massa Corporal , Comportamento de Escolha/efeitos dos fármacos , Comportamento Alimentar/efeitos dos fármacos , Hábitos , Saúde , Boca/fisiologia , Ácido Oleico/farmacologia , Paladar/fisiologia , Administração Oral , Adulto , Feminino , Humanos , Japão , Masculino , Autorrelato , Limiar Sensorial/efeitos dos fármacos , Inquéritos e Questionários , Adulto Jovem
8.
Tohoku J Exp Med ; 240(2): 153-165, 2016 10.
Artigo em Inglês | MEDLINE | ID: mdl-27760896

RESUMO

The taste detection system for oral fatty acid may be related to obesity. In addition, sleep is intrinsically and closely related to food intake and metabolism. However, the association of gustatory salivation with body mass index (BMI), daytime sleepiness, or sleep habits is largely unknown. Therefore, we evaluated the relationship between gustatory salivation and BMI, Epworth sleepiness scale (ESS, a daytime sleepiness scale) or sleep habits among 26 healthy young individuals (20 males and 6 females; mean age: 26.0 ± 4.3 years). We also measured the saliva flow rate (SFR) that was induced by gum-chewing or each of three prototypical tastants (acetic acid, sucrose, and NaCl). Further, the SFR was induced by fatty acid, provided as oleic acid (OA) homogenized in non-fat milk. All participants showed normal rates of salivation during resting and gum-chewing states. The increase in the SFR induced by OA, but not by each of the three tastants, was associated with BMI. Moreover, both daytime sleepiness level and frequency of snoring were associated with the increase in the SFR induced by NaCl. These results suggest that BMI is associated with salivation induced by oral fatty acid exposure. Additionally, the regulatory mechanism for the NaCl-induced salivation reflex may have a relationship with impairments of the respiratory control system that are related to snoring during sleep and lead to daytime sleepiness because of insufficient sleep. Thus, measurement of gustatory salivation might contribute to the evaluation and prevention of obesity and sleep-related breathing disorders.


Assuntos
Índice de Massa Corporal , Saúde , Salivação , Sono/fisiologia , Ronco/fisiopatologia , Paladar/fisiologia , Adulto , Feminino , Humanos , Masculino , Mastigação/efeitos dos fármacos , Ácido Oleico/farmacologia , Descanso , Reologia/efeitos dos fármacos , Salivação/efeitos dos fármacos , Sono/efeitos dos fármacos , Cloreto de Sódio/farmacologia , Sacarose/farmacologia , Inquéritos e Questionários , Paladar/efeitos dos fármacos , Adulto Jovem
9.
Cell Death Discov ; 10(1): 74, 2024 Feb 12.
Artigo em Inglês | MEDLINE | ID: mdl-38346947

RESUMO

Overactivation of poly (ADP-ribose) polymerase-1 (PARP-1) triggers a noncanonical form of programmed cell death (PCD) called parthanatos, yet the mechanisms of its induction are not fully understood. We have recently demonstrated that the aggresome-like induced structures (ALIS) composed of the autophagy receptor SQSTM1/p62 and K48-linked polyubiquitinated proteins (p62-based ALIS) mediate parthanatos. In this study, we identified the D1 dopamine receptor agonist YM435 as a unique parthanatos inhibitor that acts as the disaggregating agent for the p62-based ALIS. We found that YM435 structurally reduces aggregability of the ALIS, and then increases its hydrophilicity and liquidity, which prevents parthanatos. Moreover, dopamine and L-DOPA, a dopamine precursor, also prevented parthanatos by reducing the aggregability of the ALIS. Together, these observations suggest that aggregability of the p62-based ALIS determines the sensitivity to parthanatos, and the pharmacological properties of YM435 that reduces the aggregability may be suitable for therapeutic drugs for parthanatos-related diseases such as neurodegenerative diseases.

10.
Cell Struct Funct ; 38(1): 15-20, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23318214

RESUMO

We examined the regulation of neuritogenesis by a pulsed electromagnetic field (PEMF) in rat PC12 pheochromocytoma cells, which can be induced to differentiate into neuron-like cells with elongated neurites by inducers such as nerve growth factor (NGF). Plated PC12 cells were exposed to a single PEMF (central magnetic flux density, 700 mT; frequency, 0.172 Hz) for up to 12 h per day and were then evaluated for extent of neuritogenesis or acetylcholine esterase (AChE) activity. To analyze the mechanism underlying the effect of the PEMF on the cells, its effects on intracellular signaling were examined using the ERK kinase (MEK) inhibitors PD098059 and U0126 (U0124 was used as a negative control for U0126). The number of neurite-bearing PC12 cells and AChE activity increased after PEMF exposure without the addition of other inducers of neuritogenesis. Additionally, PEMF exposure induced sustained activation of ERK1/2 in PC12 cells, but not in NR8383 rat alveolar macrophages. Furthermore, U0126 strongly inhibited PEMF-dependent ERK1/2 activation and neuritogenesis. The PEMF-dependent neuritogenesis was also suppressed by PD098059, but not U0124. These results suggest that PEMF stimulation independently induced neuritogenesis and that activation of MEK-ERK1/2 signaling was induced by a cell-type-dependent mechanism required for PEMF-dependent neuritogenesis in PC12 cells.


Assuntos
Diferenciação Celular , Fator de Crescimento Neural , Neuritos , Animais , Butadienos/farmacologia , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/genética , Diferenciação Celular/efeitos da radiação , Campos Eletromagnéticos , Flavonoides/farmacologia , Humanos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/efeitos da radiação , Macrófagos Alveolares/metabolismo , Macrófagos Alveolares/efeitos da radiação , Fator de Crescimento Neural/efeitos dos fármacos , Fator de Crescimento Neural/metabolismo , Fator de Crescimento Neural/efeitos da radiação , Neuritos/efeitos dos fármacos , Neuritos/fisiologia , Neuritos/efeitos da radiação , Nitrilas/farmacologia , Células PC12 , Ratos
11.
Genes Cells ; 16(11): 1121-32, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-21988724

RESUMO

In this study, we investigated the effect of dorsomorphin, a selective inhibitor of bone morphogenetic protein (BMP) signaling, on rat PC12 pheochromocytoma cell differentiation. PC12 cells can be induced to differentiate into neuron-like cells possessing elongated neurites by nerve growth factor, BMP2, and other inducers. Cells were incubated with BMP2 and/or dorsomorphin, and the extent of neurite outgrowth was evaluated. Unexpectedly, BMP2-mediated neuritogenesis was not inhibited by co-treatment with dorsomorphin. We also found that treatment with dorsomorphin alone, but not another BMP signaling inhibitor, LDN-193189, induced neurite outgrowth in PC12 cells. To further understand the mechanism of action of dorsomorphin, the effects of this drug on intracellular signaling were investigated using the following signaling inhibitors: the ERK kinase (MEK) inhibitor U0126; the tropomyosin-related kinase A inhibitor GW441756; and the protein kinase A (PKA) inhibitor H89. Dorsomorphin induced rapid and sustained ERK1/2 activation; however, dorsomorphin-mediated ERK1/2 activation and neuritogenesis were robustly inhibited in the presence of U0126 or H89, but not GW441756. These findings suggest that dorsomorphin has the potential to induce neuritogenesis in PC12 cells, a response that requires the activation of PKA-dependent MEK-ERK1/2 signaling.


Assuntos
Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Sistema de Sinalização das MAP Quinases , Neuritos/efeitos dos fármacos , Neuritos/fisiologia , Pirazóis/farmacologia , Pirimidinas/farmacologia , Animais , Proteínas Morfogenéticas Ósseas/antagonistas & inibidores , Butadienos/farmacologia , Proteínas Quinases Dependentes de AMP Cíclico/antagonistas & inibidores , Relação Dose-Resposta a Droga , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Isoquinolinas/farmacologia , MAP Quinase Quinase Quinases/antagonistas & inibidores , MAP Quinase Quinase Quinases/metabolismo , Neuritos/enzimologia , Nitrilas/farmacologia , Células PC12 , Proteínas Quinases/metabolismo , Ratos , Transdução de Sinais/efeitos dos fármacos , Sulfonamidas/farmacologia , Ativação Transcricional/efeitos dos fármacos
12.
J Mater Sci Mater Med ; 23(10): 2461-9, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22890516

RESUMO

We prepared iron nanoparticle-encapsulating silica (FeSi) microspheres and tested their suitability as thermal seeds for hyperthermia in cancer therapy. These microspheres were prepared by introducing a ferric ion (Fe(3+)) into microspheres of a SiO(2) gel matrix derived from the hydrolysis of tetramethoxysilane in a water-in-oil emulsion that was then heat-treated at 850 °C in an argon atmosphere. The particles obtained were 5-30 µm in size and had a saturation magnetization up to 21 emu g(-1) and a coercive force of 86-133 Oe. Heat generation in an alternating current magnetic field of 300 Oe at 100 kHz was estimated to be 7.7-28.9 W g(-1). The in vitro cell biocompatibility of the microspheres was assessed by culturing rat fibroblast Rat-1 cells in medium supplemented with microspheres containing 6.7 % of iron nanoparticles. At microsphere concentrations of <7.5 g L(-1) proliferation of Rat-1 cells was not significantly inhibited.


Assuntos
Materiais Biocompatíveis , Géis , Hipertermia Induzida , Ferro/química , Nanopartículas Metálicas , Microesferas , Neoplasias/terapia , Dióxido de Silício/química , Cristalografia por Raios X , Humanos , Técnicas In Vitro , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão
13.
Nutrients ; 14(7)2022 Mar 29.
Artigo em Inglês | MEDLINE | ID: mdl-35406043

RESUMO

The Gly16Arg polymorphism results in a G to C nucleotide mutation in the human beta 2-adrenergic receptor (ADRB2) gene and has a relationship with obesity; however, this substitution's effects on food preferences are unclear. Therefore, we determined this relationship among healthy young adults (mean age, 23.4; n = 52). To evaluate food preferences, four categories of food (sweet, salty, sour, and bitter) along with high-fat foods were evaluated using a self-reporting questionnaire. Male (n = 26) and female subjects (n = 26) were genotyped for the polymorphism and further divided into three groups (two homozygous groups, GG, CC; and a heterozygous group, GC). Preference for sour foods in the GG group was higher compared with that in the CC group in females (p < 0.05). When sweet foods were classified into low- and high-fat subgroups, preference for high-fat sweet foods in the GG group was higher than that for low-fat sweet foods in all subjects (p < 0.05). The degree of preference for high-fat foods in the GG group was higher than other groups for males (p < 0.05). These results suggest that ADRB2 polymorphism is associated with food preference. Understanding the relationship of ADRB2 substitution to food preference will be valuable for designing individualized anti-obesity strategies.


Assuntos
Preferências Alimentares , Receptores Adrenérgicos beta 2 , Paladar , Adulto , Feminino , Humanos , Japão , Masculino , Obesidade , Receptores Adrenérgicos beta 2/genética , Paladar/genética , Percepção Gustatória/genética , Adulto Jovem
14.
Tohoku J Exp Med ; 222(2): 121-9, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20919447

RESUMO

Bone morphogenetic proteins (BMPs), members of the transforming growth factor ß cytokine superfamily, elicit various biological effects in different tissues. BMP receptor type II (BMPRII) contains a unique carboxyl-terminal region that interacts with multiple signaling molecules. However, expression of endogenous BMPRII is low in various mammalian cell lines, which hampers the analysis of BMP signaling. Therefore, we established a human cell line expressing BMPRII tagged with a Flag epitope (BMPRII-Flag) using the tetracycline-controlled Flp-In T-REx gene expression system. The BMPRII-Flag gene was introduced into the Flp-In T-REx 293 (FT293) cell line, a derivative of human 293 embryonic kidney fibroblasts. Then we analyzed the expression of key BMP target genes, inhibitors of DNA binding (Id) family members (Id1, Id2, and Id3) and the inhibitory Smads Smad6 and Smad7, in parental FT293 cells and an established cell line, FT293-BMPRII, by quantitative real-time PCR. Tetracycline treatment significantly increased the expression of BMPRII-Flag mRNA and protein in FT293-BMPRII cells, but induced no significant changes in expression of Id1, Id2, Id3, Smad6, or Smad7 mRNA. In contrast, treatment with a BMPRII ligand BMP2 induced the expression of Id1, Id2, Id3, and Smad6 in parental FT293 cells and FT293-BMPRII cells. Tetracycline-induced BMPRII-Flag expression significantly enhanced the induction of Id1, Id3, and Smad6 mRNA expression in FT293-BMPRII cells treated with BMP2. These findings provide evidence that although BMPRII has no obvious effect on the expression of representative BMP target genes, it differentially modulates the responsiveness of target genes to BMP2.


Assuntos
Proteína Morfogenética Óssea 2/metabolismo , Receptores de Proteínas Morfogenéticas Ósseas Tipo II/metabolismo , Fibroblastos/metabolismo , Receptores de Proteínas Morfogenéticas Ósseas Tipo II/genética , Fibroblastos/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Células HEK293 , Humanos , Proteína 1 Inibidora de Diferenciação/genética , Proteína 1 Inibidora de Diferenciação/metabolismo , Proteínas Inibidoras de Diferenciação/genética , Proteínas Inibidoras de Diferenciação/metabolismo , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Oligopeptídeos , Peptídeos/metabolismo , Inibidores da Síntese de Proteínas/farmacologia , RNA Mensageiro/metabolismo , Proteínas Recombinantes , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais/efeitos dos fármacos , Proteína Smad6/genética , Proteína Smad6/metabolismo , Tetraciclina/farmacologia
15.
J Craniofac Surg ; 21(6): 1839-42, 2010 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-21119436

RESUMO

This study was undertaken to examine effects and biocompatibility of a new internalized distraction device made from newly developed Ti-Nb-Al shape memory alloy (SMA). Crania of Wistar rats were expanded using a U-shaped wire of this SMA set on each cranium in an experimental group. At 2 or 4 weeks after operation, the rats were killed; width measurements and three-dimensional observations of crania were conducted using soft x-ray and microfocus x-ray computed tomography photography. After photography, histologic sections were made and stained with hematoxylin and eosin. No pathologic change in the experimental duration was observed macroscopically or histologically. Significantly increased size was found for the rat crania in the experimental group compared with the control group. Results demonstrated the feasibility and biocompatibility of internalized distraction osteogenesis using Ni-free, Ti-based SMA in craniofacial plastic surgery for craniofacial deformities.


Assuntos
Ligas/química , Materiais Biocompatíveis/química , Ligas Dentárias/química , Osteogênese por Distração/instrumentação , Osso Parietal/cirurgia , Procedimentos de Cirurgia Plástica/instrumentação , Animais , Cefalometria/métodos , Corantes , Tecido Conjuntivo/diagnóstico por imagem , Tecido Conjuntivo/patologia , Suturas Cranianas/diagnóstico por imagem , Suturas Cranianas/patologia , Amarelo de Eosina-(YS) , Desenho de Equipamento , Estudos de Viabilidade , Corantes Fluorescentes , Hematoxilina , Imageamento Tridimensional/métodos , Fixadores Internos , Masculino , Osso Parietal/diagnóstico por imagem , Osso Parietal/patologia , Fotografação/métodos , Ratos , Ratos Wistar , Fatores de Tempo , Microtomografia por Raio-X/métodos
16.
J Biomed Mater Res B Appl Biomater ; 105(8): 2308-2314, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27495744

RESUMO

Magnetic microspheres measuring 15-35 µm in diameter are believed to be useful for intra-arterial hyperthermia. In this study, we attempted to prepare titanium dioxide (TiO2 ) microspheres containing magnetic nanoparticles (MNPs). MNP-containing TiO2 microspheres with diameters of approximately 30 µm were successfully obtained by sol-gel reaction of titanium tetraisopropoxide in a water-in-oil emulsion with added cosurfactant of 1-butanol and subsequent heat treatment at 200°C. The microspheres showed ferrimagnetism owing to high content of MNPs in approximately 60 wt % and had a low-crystalline TiO2 matrix. Furthermore, the agar phantom was heated to above 43°C after approximately 1 min under an alternating magnetic field of 100 kHz and 300 Oe and showed in vitro biocompatibility similar to that of MNP-free TiO2 microspheres. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 2308-2314, 2017.


Assuntos
Hipertermia Induzida/métodos , Campos Magnéticos , Nanopartículas de Magnetita/química , Titânio , Animais , Linhagem Celular , Ratos , Titânio/química , Titânio/farmacologia
17.
Mater Sci Eng C Mater Biol Appl ; 69: 1268-72, 2016 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-27612826

RESUMO

Initial cellular responses following implantation are important for inducing osteoconduction. We investigated cell adhesion, spreading, proliferation and differentiation of mouse MC3T3-E1 osteoblastic cells on untreated or fibronectin (Fn)-coated discs of hydroxyapatite (HAp) or alpha-type alumina (α-Al2O3). Fn coating significantly enhanced adhesion and spreading of MC3T3-E1 cells on HAp, but did not affect MC3T3-E1 cell proliferation and differentiation on HAp or α-Al2O3. Fn-coated HAp likely does not stimulate pre-osteoblast cells to initiate the process of osteoconduction; however, Fn adsorption might affect the response of inflammatory cells to the implanted material or, in conjunction with other serum proteins, stimulate pre-osteoblast cell proliferation and differentiation. Further studies on the effect of serum proteins in cell culture and the efficacy of Fn-coated HAp and α-Al2O3in vivo are warranted.


Assuntos
Óxido de Alumínio/farmacologia , Durapatita/farmacologia , Fibronectinas/farmacologia , Osteoblastos/citologia , Adsorção , Fosfatase Alcalina/metabolismo , Animais , Animais Recém-Nascidos , Bovinos , Adesão Celular/efeitos dos fármacos , Contagem de Células , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Camundongos Endogâmicos C57BL , Microscopia de Fluorescência , Osteoblastos/efeitos dos fármacos
18.
Biomed Mater ; 11(4): 045006, 2016 08 10.
Artigo em Inglês | MEDLINE | ID: mdl-27509476

RESUMO

The osteoconductivity mechanism of hydroxyapatite (HAp) has not been elucidated. It is hypothesized that specific proteins adsorb on HAp, promoting its osteoconductivity. To verify this hypothesis, we compared the adsorption behavior of fibronectin (Fn) on HAp powder and on α-alumina (α-Al2O3) powder, a material with no osteoconductivity. More Fn adsorbed on α-Al2O3 than on HAp, irrespective of the Fn concentration, and there was no significant difference in the secondary structure of Fn adsorbed on HAp and α-Al2O3. Further, it is possible that Fn did not adsorb on HAp and α-Al2O3 through the Arg-Gry-Asp motif of Fn. The amount of Fn adsorbed on HAp oriented to the a(b)-axis with very little decrease in carbonate and the adsorbed Fn had a smaller α-helix structure content. The results suggest that the secondary and/or higher-order structure rather than the amount of adsorbed Fn might affect the osteoconductivity of HAp, which might be electrostatically controlled by the crystal face orientation and/or carbonate content of HAp, although this should be confirmed by a cell culture test in the future.


Assuntos
Óxido de Alumínio/química , Regeneração Óssea , Durapatita/química , Fibronectinas/química , Adsorção , Substitutos Ósseos , Adesão Celular , Humanos , Microscopia Eletrônica de Varredura , Oligopeptídeos/química , Pós , Domínios Proteicos , Estrutura Secundária de Proteína , Espectroscopia de Infravermelho com Transformada de Fourier , Eletricidade Estática , Propriedades de Superfície , Difração de Raios X
19.
ACS Biomater Sci Eng ; 2(7): 1162-1168, 2016 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-33465874

RESUMO

Artificial hydroxyapatite (HAp) is osteoconductive, but the mechanism is still unclear. It is likely that some serum proteins are adsorbed onto HAp and influence its osteoconductivity. We investigated the adsorption behavior of laminin (LN), which was isolated from murine Engelbreth-Holm-Swarm sarcoma, onto HAp and compared it with nonosteoconductive alpha-type alumina (α-Al2O3). Cell adhesion, spreading, and proliferation on native and LN-adsorbed discs of HAp or α-Al2O3 were examined using murine MC3T3-E1 osteoblastic cells. A larger amount of LN adsorbed onto HAp than α-Al2O3 despite the electrostatic repulsion between LN and HAp, suggesting the specific adsorption of LN onto HAp. The LN adsorbed onto HAp remarkably enhanced initial attachment and spreading of MC3T3-E1 cells, but subsequent proliferation of MC3T3-E1 cells was influenced by the type of material rather than LN adsorption. These fundamental findings imply that LN adsorbed on HAp could trigger osteoconductivity in vivo, aiding in the development of novel biomaterials that specifically adsorb LN and effectively enhance cell attachment and spreading.

20.
J Biomater Appl ; 30(1): 119-30, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25659946

RESUMO

Shape memory alloys (SMAs) including superelastic alloys have unique properties such as shape memory and superelasticity, thus they are recognized as very useful biomaterials. These properties are very advantageous for medical use, and actually the SMA wires have been widely used in medical field. However, biocompatibility of nickel-titanium (Ni-Ti) alloy, which is the only practical SMA at present, has been questioned because of its high nickel content. The aim of this study was to evaluate the biocompatibility of a newly developed Ni-free Ti-based SMA for medical use. The newly developed SMA made of Ti-Mo-Sn-Zr system was processed into a disk of 15.1 mm in diameter. Pure titanium of the same shape was prepared as control. All the disk surfaces were polished using emery papers, #120, #400, and #600. Scanning electron microscopy and a 3D optics profiler were used to evaluate the surface of the materials. In vitro evaluations included colony examination for evaluation of the cell cytotoxicity, DNA quantification for the cell proliferation, Alamar blue assay for metabolic activity, FDA staining for the live cell imaging, and cell cycle analysis, using Chinese hamster fibroblastic V-79 cells and mouse osteoblastic MC3T3-E1 cells. In colony examination and DNA quantification, there was no significant difference between the Ti-Mo-Sn-Zr and the pure titanium. In FDA staining, cultured cells on the Ti-Mo-Sn-Zr alloy showed the same biocompatibility as those on the pure titanium. The present results suggest that the newly developed Ti-Mo-Sn-Zr alloy showed the high biocompatibility comparable to pure titanium and can be used as efficient biomaterial for medical use.


Assuntos
Ligas/química , Materiais Biocompatíveis/química , Molibdênio/química , Estanho/química , Titânio/química , Zircônio/química , Ligas/toxicidade , Animais , Materiais Biocompatíveis/toxicidade , Ciclo Celular/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Cricetinae , DNA/análise , Elasticidade , Camundongos , Molibdênio/toxicidade , Estresse Mecânico , Estanho/toxicidade , Titânio/toxicidade , Zircônio/toxicidade
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