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1.
Int J Biol Macromol ; 185: 1022-1035, 2021 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-34197859

RESUMO

Biochemical modification can endow the surface of implants with superior biological activity. Herein, silk fibroin (SF) protein and its anionic derivative peptides (Cs) were covalently immobilized onto a titanium implant surface via a polydopamine layer. The successful conjugation of SF and Cs was revealed by X-ray photoelectron spectroscopy (XPS), field-emission scanning electron microscopy (FE-SEM), atomic force microscopy (AFM), and contact angle measurements. The addition of Cs prevented the conformational transition of silk fibroin to silk II. The deposition of apatite on its surface was significantly accelerated, and the bioactive composite coating was observed to enhance protein adsorption and cell proliferation. More importantly, it also promoted the osteogenic differentiation of bone marrow stem cells (BMSCs) for the quantitative and qualitative detection of alkaline phosphatase (ALP) and alizarin red (ARS). Overall, the stable performance and enhanced osteogenic property of the composite coating promote an extensive application for clinical titanium-based implants.


Assuntos
Fibroínas/farmacologia , Indóis/química , Células-Tronco Mesenquimais/citologia , Osteogênese/efeitos dos fármacos , Polímeros/química , Titânio/química , Adsorção , Animais , Apatitas/química , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Fibroínas/química , Células-Tronco Mesenquimais/efeitos dos fármacos , Camundongos , Microscopia de Força Atômica , Oxirredução , Peptídeos/química , Peptídeos/farmacologia , Espectroscopia Fotoeletrônica
2.
Biomed Mater ; 16(4)2021 06 25.
Artigo em Inglês | MEDLINE | ID: mdl-34098538

RESUMO

In this study, the silk fibroin/nano-hydroxyapatite/hyaluronic acid (SF/nHAp/HA) composite scaffolds with different HA contents were developed by blending, cross-linking and freeze-drying, and their physicochemical properties and cell biocompatibilityin vitrowere subsequently studied. It was observed that the molecular conformation of the composite scaffolds was mainly composed of silk I and a small amount of theß-sheets structure. On enhancing the HA content, the pore size of the scaffold decreased, while the porosity, water absorption, swelling ratio and mechanical properties were observed to increase. In particular, the SF/nHAp/HA scaffold with a 5.0 wt% ratio exhibited the highest water absorption and mechanical properties among the developed materials. In addition, thein vitrocytocompatibility analysis showed that the bone marrow mesenchymal stem cells exhibited excellent cell proliferation and osteogenic differentiation ability on the SF/nHAp/5.0 wt%HA scaffolds, as compared with the other scaffolds. It can be concluded that the developed composite scaffolds represent a promising class of materials for the bone tissue repair and regeneration.

3.
Transl Pediatr ; 10(2): 366-377, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33708523

RESUMO

Background: In responding to the potential challenges in the transition from childhood to adulthood for individuals with autism spectrum disorders (ASD) in China, a novel conception of "ALSO" was proposed to bridge the transitional needs and early intervention. To facilitate the application of ALSO in early intervention, ALSOLIFE skills assessment system (ALSOLIFE Assessment) was developed to enable caregivers to evaluate their children's skills guided by the ALSO conception. Given that the critical shortage of qualified professionals in China, many caregivers of children with ASD must function as home therapists. To address the practical needs of Chinese families of children with ASD, ALSOLIFE Assessment is designed as a free, online, technology-assisted, self-operated and behavioral intervention approaches supported system. The assessment report then further served as the basis for caregivers to deliver the tailored educational intervention to their children. Although ALSOLIFE Assessment provides caregivers a home-based intervention program, it is still unclear whether its evaluation is reliable and accurate. Therefore, we conduct this study to investigate the reliability and validity of the ALSOLIFE Assessment. Methods: A total of 1,050 children with ASD (1 to 10 years old) were recruited from 31 provinces of mainland China. Their caregivers participated simultaneously as ALSOLIFE Assessment evaluators. The testing results of Verbal Behavior Milestones Assessment and Placement Program (VB-MAPP) and Psychoeducational Profile-3 (PEP-3) were also collected for a portion of the child participants (VB-MAPP, N=34; PEP-3, N=31) to check criterion-related validity. Results: The reliability and validity of the ALSOLIFE Assessment satisfied psychometric requirements after the reduction from 511 to 464 items. Exploratory factor analysis (EFA) of ALSOLIFE Assessment scoring data yielded six factors, and confirmatory factor analysis (CFA) verified the best fitting construct model is a bifactorial model with one general factor and six group factors. Compared to the VB-MAPP and PEP-3, the ALSOLIFE Assessment exhibited good criterion-related validity across three levels: total scores, 6 skill domains, and 22 skill subdomains. The reliability tests indicated its strong internal consistency, test-retest reliability, and inter-rater reliability. Conclusions: The reliability and validity of the revised ALSOLIFE Assessment (464-items) satisfied psychometric requirements. It is essential to develop validated and comprehensive evaluation tools, embedded on the needs of Chinese families of children with ASD.

4.
J Stroke Cerebrovasc Dis ; 28(2): 243-250, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30503677

RESUMO

BACKGROUND: Recently, the roles of ß-fibrinogen (FGß) polymorphisms in ischemic stroke (IS) were intensively analyzed, but the results of these studies were inconsistent. Thus, we performed this study to better assess potential relationship between FGß polymorphisms and the risk of IS. METHODS: Eligible studies were searched in PubMed, Medline, Embase, and CNKI. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated to evaluate correlations between FGß polymorphisms and IS. RESULTS: A total of 49 studies were included for analyses. Significant associations with the risk of IS were detected for FGß -148 C/T and -455 G/A polymorphisms in overall analyses. Further subgroup analyses according to ethnicities of participants revealed that the -148 C/T polymorphism was significantly correlated with the risk of IS in both Asians and Caucasians, while the -455 G/A polymorphism was only significantly correlated with the risk of IS in Asians. When we stratified available data according to types of disease, we found that both FGß -148 C/T and -455 G/A polymorphisms were significantly correlated with the risk of cerebral infarction. CONCLUSIONS: Our findings indicated that FGß -148 C/T and -455 G/A polymorphisms may serve as potential biological markers for IS in Asians. Moreover, the FGß -148 C/T polymorphism may also serve as a potential biological marker for IS in Caucasians.


Assuntos
Isquemia Encefálica/genética , Fibrinogênio/genética , Polimorfismo Genético , Acidente Vascular Cerebral/genética , Grupo com Ancestrais do Continente Asiático/genética , Isquemia Encefálica/diagnóstico , Isquemia Encefálica/etnologia , Estudos de Casos e Controles , Grupo com Ancestrais do Continente Europeu/genética , Frequência do Gene , Estudos de Associação Genética , Predisposição Genética para Doença , Humanos , Fenótipo , Fatores de Risco , Acidente Vascular Cerebral/diagnóstico , Acidente Vascular Cerebral/etnologia
5.
J Org Chem ; 81(10): 4310-5, 2016 05 20.
Artigo em Inglês | MEDLINE | ID: mdl-27128964

RESUMO

We described the first total syntheses of clausenapin, indizoline, claulansine M, and a novel synthetic route to clausenaline D via divergent method. Key steps involved TFAA-mediated intramolecular acylation to construct the carbazole core and subsequent Claisen rearrangement to generate key intermediates for further elaboration to target molecules.


Assuntos
Alcaloides/síntese química , Carbazóis/síntese química , Acilação , Indicadores e Reagentes , Alcaloides Indólicos , Espectroscopia de Ressonância Magnética , Estereoisomerismo
6.
J Acupunct Meridian Stud ; 7(5): 225-30, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25441946

RESUMO

The current study investigated the effects of acupuncture at Zu-San-Li (ST36) on the hypothalamic-pituitary-adrenal axis during ethanol withdrawal in rats. Rats were intraperitoneally treated with 3 g/kg/day of ethanol or saline for 28 days. Following 24 hours of ethanol withdrawal, acupuncture was applied at bilateral ST36 points or non-acupoints (tail) for 1 minute. Plasma levels of corticosterone (CORT) and adrenocorticotropic hormone (ACTH) were measured by radioimmunoassay (RIA), and the corticotropin-releasing factor (CRF) protein levels in the paraventricular nucleus of the hypothalamus were also examined by RIA 20 minutes after the acupuncture treatment. RIA showed significantly increased plasma levels of CORT and ACTH in the ethanol-withdrawn rats compared with the saline-treated rats, which were inhibited significantly by the acupuncture at the acupoint ST36 but not at the non-acupoint. Additionally, ethanol withdrawal promoted CRF protein expressions in the paraventricular nucleus of the hypothalamus, which were also blocked by the acupuncture at ST36. These findings suggest that acupuncture at the specific acupoint ST36 can inhibit ethanol withdrawal-induced hyperactivation of hypothalamic-pituitary-adrenal axis, and it may be mediated via the modulation of hypothalamic CRF.


Assuntos
Pontos de Acupuntura , Terapia por Acupuntura , Etanol/efeitos adversos , Sistema Hipotálamo-Hipofisário/fisiologia , Sistema Hipófise-Suprarrenal/fisiologia , Síndrome de Abstinência a Substâncias/terapia , Hormônio Adrenocorticotrópico/sangue , Animais , Corticosterona/sangue , Hormônio Liberador da Corticotropina/sangue , Masculino , Ratos , Ratos Sprague-Dawley , Síndrome de Abstinência a Substâncias/sangue , Síndrome de Abstinência a Substâncias/metabolismo
7.
Biosci Biotechnol Biochem ; 78(7): 1159-66, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25229851

RESUMO

Skeletal and cardiac muscle have important roles in glucose uptake and utilization. However, changes in expression of protein coding genes and miRNAs that participate in glucose metabolism during development are not fully understood. In this study, we investigated the expression of genes related to glucose metabolism during muscle development. We found an age-dependent increase in gene expression in cardiac muscle, with enrichment in heart development- and energy-related metabolic processes. A subset of genes that were up-regulated until 30 or 180 days postnatally, and then down-regulated in psoas major muscle was significantly enriched in mitochondrial oxidative-related processes, while genes that up-regulated in longissimus doris muscle was significantly enriched in glycolysis-related processes. Meanwhile, expression of energy-related microRNAs decreased with increasing age. In addition, we investigated the correlation between microRNAs and mRNAs in three muscle types across different stages of development and found many potential microRNA-mRNA pairs involved in regulating glucose metabolism.


Assuntos
Glucose/metabolismo , Coração/crescimento & desenvolvimento , Desenvolvimento Muscular/genética , Músculo Esquelético/crescimento & desenvolvimento , Suínos , Transcriptoma , Animais , Transporte Biológico/genética , Metabolismo Energético/genética , Feminino , MicroRNAs/genética , Músculo Esquelético/metabolismo , Miocárdio/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
8.
J Clin Neurosci ; 21(2): 246-9, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24239227

RESUMO

Some polymorphisms in the fibroblast growth factor receptor 4 gene (FGFR-4) have been correlated with coronary artery disease, however, the role of polymorphisms in the FGFR-4 gene in ischemic stroke remain unknown. A total of 270 patients with ischemic stroke and 297 controls were recruited. Stroke subtype was classified and clinical severity of stroke in patients was evaluated. The polymorphisms in the FGFR-4 were genotyped. There were no significant differences of genotype distributions and allele frequencies of rs145302848C/G and rs147603016G/A between stroke patients and controls (all p>0.05). However, genotype frequencies and allele frequencies at rs351855G/A (Gly388Arg) were significantly different between stroke patients and controls (both p<0.001). With the rs351855GG genotype as a reference, the presence of rs351855AA homozygote had a significantly increased risk for stroke (adjusted odds ratio 2.663; 95% confidence interval 1.673-4.229, p<0.001). The polymorphisms at rs145302848C/G and rs147603016G/A did not influence the susceptibility of stroke in this study. All FGFR-4 polymorphisms were not associated with clinical features such as Trial of Org 10172 in Acute Stroke Treatment subtype or stroke severity as indicated by mean National Institutes of Health Stroke Scale scores. Our study suggests a positive association between FGFR-4 gene polymorphism at rs351855G/A and susceptibility to ischemic stroke.


Assuntos
Isquemia Encefálica/genética , Predisposição Genética para Doença , Polimorfismo Genético , Receptor Tipo 4 de Fator de Crescimento de Fibroblastos/genética , Acidente Vascular Cerebral/genética , Grupo com Ancestrais do Continente Asiático/genética , Isquemia Encefálica/fisiopatologia , China/epidemiologia , Feminino , Frequência do Gene , Estudos de Associação Genética , Genótipo , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Razão de Chances , Fatores de Risco , Índice de Gravidade de Doença , Acidente Vascular Cerebral/fisiopatologia
9.
Org Biomol Chem ; 12(5): 758-64, 2014 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-24337052

RESUMO

Ruthenium-catalyzed regioselective oxidative cross-coupling/annulations of quinazolones with alkynes were successfully developed for direct access to fused polycyclic heteroarenes. The transformation proceeded well with a broad substrate scope under mild conditions to achieve moderate to high yields.

10.
Chin J Cancer Res ; 25(5): 520-6, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24255575

RESUMO

OBJECTIVE: To test the effects of salidroside on formation and growth of glioma together with tumor microenvironment. METHODS: Salidroside extracted from Rhodiola rosea was purified and treated on human glioma cells U251 at the concentration of 20 µg/mL. 3-(4,5-dimethylthiazol-2-yl)-2,5-dephenyltetrazolium bromide (MTT) assay for cytotoxicity and flow cytometry (FCM) for cell cycle analysis were performed. Then for in vivo study, xenotransplantation tumor model in nude mice was generated and treated with salidroside at the concentration of 50 mg/kg(.)d for totally 20 d. Body weight and tumor size were detected every 2 d after the treatment. The levels of 8-isoprostane, superoxide dismutase (SOD) and malondialdehyde (MDA), special markers for oxidative stress, were detected while immunofluoresence staining was performed for astrocyte detection. RESULTS: For in vitro study, salidroside could decrease the viability of human glioma cells U251 and the growth of U251 cells at G0/G1 checkpoint during the cell cycle. For in vivo study, salidroside could also inhibit the growth of human glioma tissue in nude mice. The body weight of these nude mice treated with salidroside did not decrease as quickly as control group. In the tumor xenotransplantation nude mice model, mice were found of inhibition of oxidative stress by detection of biomarkers. Furthermore, overgrowth of astrocytes due to the stimulation of oxidative stress in the cortex of brain was inhibited after the treatment of salidroside. CONCLUSIONS: Salidroside could inhibit the formation and growth of glioma both in vivo and in vitro and improve the tumor microenvironment via inhibition of oxidative stress and astrocytes.

11.
Int J Biol Sci ; 9(6): 550-6, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23781149

RESUMO

Functional differences in the different types of adipose tissue and the impact of their dysfunction on metabolism are associated with the regional distribution of adipose depots. Here we show a genome-wide comparison between the transcriptomes of one source of subcutaneous and two sources of visceral adipose tissue in the pig using an RNA-seq approach. We obtained ~32.3 million unique mapped reads which covered ~80.2% of the current annotated transcripts across these three sources of adipose tissue. We identified various genes differentially expressed between subcutaneous and visceral adipose tissue, which are potentially associated with the inflammatory features of visceral adipose tissue. These results are of benefit for understanding the phenotypic, metabolic and functional differences between different types of adipose tissue that are deposited in different body sites.


Assuntos
Inflamação/patologia , Gordura Intra-Abdominal/patologia , Transcriptoma , Animais , Ácidos Graxos/metabolismo , Feminino , Inflamação/genética , Gordura Intra-Abdominal/metabolismo , Análise de Sequência de RNA , Suínos
12.
PLoS One ; 8(3): e59440, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23555673

RESUMO

Compared to normal aging adults, individuals with amnestic mild cognitive impairment (aMCI) have significantly increased risk for progressing into Alzheimer's disease (AD). Autopsy studies found that most of the brains of aMCI cases showed anatomical features associated with AD pathology. The recent development of non-invasive neuroimaging technique, such as diffusion tensor imaging (DTI), makes it possible to investigate the microstructures of the cerebral white matter in vivo. We hypothesized that disrupted white matter (WM) integrity existed in aMCI. So we used DTI technique, by measuring fractional anisotropy (FA) and mean diffusivity (MD), to test the brain structures involved in patients with aMCI. DTI scans were collected from 40 patients with aMCI, and 28 normal controls (NC). Tract-based spatial statistics (TBSS) analyses of whole-brain FA and MD images in each individual and group comparisons were carried out. Compared to NC, aMCI patients showed significant FA reduction bilaterally, in the association and projection fibers of frontal, parietal, and temporal lobes, corpus callosum, bilateral corona radiation, right posterior thalamic radiation and right sagittal stratum. aMCI patients also showed significantly increased MD widespreadly in the association and projection fibers of frontal, parietal and temporal lobes, and corpus callosum. Assessment of the WM integrity of the frontal, parietal, temporal lobes, and corpus callosum by using DTI measures may aid early diagnosis of aMCI.


Assuntos
Amnésia/complicações , Encéfalo/patologia , Disfunção Cognitiva/complicações , Disfunção Cognitiva/patologia , Imagem de Tensor de Difusão , Idoso , Encéfalo/fisiopatologia , Estudos de Casos e Controles , Disfunção Cognitiva/diagnóstico , Disfunção Cognitiva/fisiopatologia , Corpo Caloso/patologia , Corpo Caloso/fisiopatologia , Feminino , Humanos , Sistema Límbico/parasitologia , Sistema Límbico/patologia , Masculino , Pessoa de Meia-Idade , Rede Nervosa/patologia , Rede Nervosa/fisiopatologia
14.
Chin Med J (Engl) ; 125(12): 2168-72, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22884148

RESUMO

BACKGROUND: Serum testosterone levels have been found lower in acute ischemic stroke male patients. However, the exact mechanism remains unclear. In the present study, we measured serum testosterone levels, steroidogenesis- related genes and Leydig cells number in experimental transient cerebral ischemia male rats to elucidate the mechanism. METHODS: The middle cerebral arteries of adult male Sprague-Dawley rats were sutured for 120 minutes and then sacrificed after 24 hours. Blood was collected for measurement of serum testosterone, follicular stimulating hormone and estradiol levels, and testes were collected for measurement of steroidogenesis-related gene mRNA levels and number of Leydig cells. RESULTS: Serum testosterone levels in rats after cerebral ischemia were significantly lower (0.53 ± 0.16) ng/ml, n = 7, mean ± SE) compared with control ((2.33 ± 0.60) ng/ml, n = 7), while serum estradiol and follicular stimulating hormone levels did not change. The mRNA levels for luteinizing hormone receptor (Lhcgr), scavenger receptor class B member 1 (Scarb1), steroidogenic acute regulatory protein (StAR), cholesterol side chain cleavage enzyme (Cyp11a1), 3ß-hydroxysteroid dehydrogenase 1 (HSD3ß1), 17α-hydroxylase/20-lyase (Cyp17a1) and membrane receptor c-kit (kit) were significantly downregulated by cerebral ischemia, while luteinizing hormone, Kit ligand (KitL), 17ß-hydrosteroid dehydrogenase 3 (HSD17ß3) and 5α-reductase (Srd5a1) were not affected. We also observed that, relative to control, the Leydig cell number did not change. CONCLUSIONS: These results indicate that transient cerebral ischemia in the brain results in lower expression levels of steroidogenesis-related genes and thus lower serum testosterone level. Transient cerebral ischemia did not lower the number of Leydig cells.


Assuntos
Ataque Isquêmico Transitório/sangue , Ataque Isquêmico Transitório/metabolismo , Testículo/metabolismo , Animais , Ensaio de Imunoadsorção Enzimática , Estradiol/sangue , Hormônio Foliculoestimulante/sangue , Células Intersticiais do Testículo/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Testosterona/sangue
15.
Int J Mol Sci ; 12(11): 7950-9, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22174642

RESUMO

MicroRNAs (miRNAs) are non-coding small RNAs that play roles in regulating gene expression. Some miRNAs have been classed as epigenetic regulators of metabolism and energy homeostasis. Previous reports indicated that the miRNAs miR-27a and miR-143 were involved in lipid metabolism in human and rodents. To determine the roles of porcine miR-27a and miR-143 in adipocyte lipid metabolism, porcine adipocytes were cultured and allowed to induce differentiation for 10 days. The lipid-filled adipocytes were then transfected with miRNA mimics and inhibitors. We measured how the indicators of adipogenesis and adipolysis in porcine adipocytes were affected by the over-expression and by the inhibition of both miR-27a and miR-143. The results indicated that the over-expression of miR-27a could accelerate adipolysis releasing significantly more glycerol and free fatty acids than the negative control (P < 0.001), while the mimic of miR-143 expression, promoted adipogenesis by accumulating more triglycerides (P < 0.001) in the adipocytes. In addition, we demonstrated that there was good correlation (r > 0.98, P < 0.001) between the indicators of adipolysis in cell lysates and in the culture medium.


Assuntos
Adipócitos/metabolismo , Metabolismo dos Lipídeos/genética , MicroRNAs/genética , Adipogenia/genética , Animais , Diferenciação Celular , Regulação da Expressão Gênica , MicroRNAs/antagonistas & inibidores , MicroRNAs/metabolismo , Suínos , Transfecção , Triglicerídeos/metabolismo
16.
BMC Genomics ; 11: 429, 2010 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-20624324

RESUMO

BACKGROUND: Currently, a number of yeast genomes with different physiological features have been sequenced and annotated, which provides invaluable information to investigate yeast genetics, evolutionary mechanism, structure and function of gene families. DESCRIPTION: YeastWeb is a novel database created to provide access to gene families derived from the available yeast genomes by assigning the genes into putative families. It has many useful features that complement existing databases, such as SGD, CYGD and Génolevures: 1) Detailed computational annotation was conducted with each entry with InterProScan, EMBOSS and functional/pathway databases, such as GO, COG and KEGG; 2) A well established user-friendly environment was created to allow users to retrieve the annotated genes and gene families using functional classification browser, keyword search or similarity-based search; 3) Workset offers users many powerful functions to manage the retrieved data efficiently, associate the individual items easily and save the intermediate results conveniently; 4) A series of comparative genomics and molecular evolution analysis tools are neatly implemented to allow users to view multiple sequence alignments and phylogenetic tree of gene families. At present, YeastWeb holds the gene families clustered from various MCL inflation values from a total of 13 available yeast genomes. CONCLUSIONS: Given the great interest in yeast research, YeastWeb has the potential to become a useful resource for the scientific community of yeast biologists and related researchers investigating the evolutionary relationship of yeast gene families. YeastWeb is available at http://centre.bioinformatics.zj.cn/Yeast/.


Assuntos
Bases de Dados Genéticas , Genes Fúngicos/genética , Internet , Leveduras/genética , Evolução Molecular , Armazenamento e Recuperação da Informação , Análise de Sequência de DNA , Interface Usuário-Computador
17.
Connect Tissue Res ; 49(2): 92-8, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18382895

RESUMO

Hypertrophic scars and keloid are dermal proliferative disorders in wound healing. Transforming growth factor beta (TGF-beta) has been implicated in scar formation through the activation of fibroblasts and the acceleration of collagen deposition. Our study aimed to design a novel truncated (27-123 residues) type II TGF-beta receptor (tTGFbetaRII) and to determine its effects on the proliferation of keloid fibroblasts and the collagen synthesis as well as TGF-beta I expression of the cells. The coding sequences of TGF-beta I and tTGFbetaRII were amplified using RT-PCR and then cloned into pGBKT7 and pGADT7 vectors. A yeast two-hybrid experiment and a glutathione S-transferase (GST)-pull down assay were performed to verify the affinity of tTGFbetaRII to TGF-beta I. Our results indicated that treatment with tTGFbetaRII inhibited the growth of keloid fibroblasts and suppressed the synthesis of type I collagen in keloid fibroblasts in a concentration-dependent manner. Moreover, northern and western blot analysis revealed a decline of the TGF-beta I expression at both mRNA and protein levels after exposure to 5, 10 or 20 mug/ml of tTGFbetaRII. Together, our data suggested that the exogenous tTGFbetaRII can efficiently trap TGF-beta I from access to wild-type receptors and can suppress TGF-beta I triggered signals. Thus it may potentially be clinically applied to scar therapy.


Assuntos
Colágeno/biossíntese , Fibroblastos/metabolismo , Queloide/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Receptores de Fatores de Crescimento Transformadores beta/metabolismo , Pele/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Relação Dose-Resposta a Droga , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/genética , Fibroblastos/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/genética , Vetores Genéticos , Humanos , Queloide/genética , Queloide/terapia , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/farmacologia , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , Receptor do Fator de Crescimento Transformador beta Tipo II , Receptores de Fatores de Crescimento Transformadores beta/genética , Saccharomyces cerevisiae , Pele/patologia , Pele/fisiopatologia , Transfecção , Fator de Crescimento Transformador beta/genética , Cicatrização/efeitos dos fármacos , Cicatrização/genética
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