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1.
J Craniofac Surg ; 32(8): 2821-2822, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34727484

RESUMO

ABSTRACT: Rhinophyma, the final stage of acne rosacea, severely influences the patient's appearance and can only be treated by surgical methods. This case reports a simple, safe, effective, and economical surgical method-five-blade scratcher. After the surgical treatment, the overall nasal contour of the patient, a male with severe rosacea, was restored without scar formation. Thus, this surgical method reported in this case is feasible and easy to operate, and worthy of clinical promotion.


Assuntos
Rinofima , Rosácea , Cicatriz , Humanos , Masculino , Nariz/cirurgia , Rinofima/cirurgia , Instrumentos Cirúrgicos
2.
Biomed Pharmacother ; 120: 109489, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31629950

RESUMO

Depression is a complicated etiological pattern, and its pathology and effective treatments are highly limited.C1q-tumor necrosis factor-related protein-3 (CTRP3) is an adipokine, playing crucial roles in metabolic regulatory properties. However, the effects of CTRP3 on depression are largely unknown. In the present study, we found that CTRP3 expression levels were markedly reduced in hippocampus of mice with depression induced by chronic unpredictable mild stress (CUMS). In mouse model with depression, CTRP3-deficient mice aggravated depression-associated behaviors, as evidenced by the reduced locomotor activity and sucrose consumption, while the elevated immobility time in the tail suspension test (TST) and forced swimming test (FST). Moreover, CUMS-induced neuron death and increased expression of cleaved Caspase-3 were significantly accelerated by CTRP3 knockout. Furthermore, CTRP3 deletion intensified pro-inflammatory response in CUMS-exposed mice, which was associated with the activation of nuclear factor-κB(NF-κB) signaling. The activity of mitogen-activated protein kinases (MAPKs), including p38 and JNK, was further promoted in hippocampus of CTRP3-knockout mice with CUMS exposure. In contrast,CTRP3 over-expression showed anti-apoptotic and anti-inflammatory effects in lipopolysaccharide (LPS)-treated microglial cells. Importantly, the in vitro experiments demonstrated that CTRP3 knockdown-exacerbated apoptosis and inflammatory responsewere remarkably abrogated by the blockage of p38 and JNK signaling pathways in microglia stimulated by LPS. Next, in CUMS-exposed mice with CTRP3 deficiency, suppressing p38 and JNK markedly alleviated depressive-like behavior,hippocampal neuron death, apoptosis and inflammation. Therefore, CTRP3 may be an innovative therapeutic target for treating patients with depression through regulating p38 and JNK signaling.


Assuntos
Adipocinas/metabolismo , Apoptose/fisiologia , Depressão/metabolismo , Transtorno Depressivo/metabolismo , Inflamação/metabolismo , Sistema de Sinalização das MAP Quinases/fisiologia , Transdução de Sinais/fisiologia , Animais , Comportamento Animal/fisiologia , Células Cultivadas , Modelos Animais de Doenças , Hipocampo/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Microglia/metabolismo , NF-kappa B/metabolismo , Estresse Psicológico/metabolismo
3.
Inflammation ; 41(2): 606-613, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29218605

RESUMO

Decreased interferon (IFN)-γ levels and increased levels of macrophage-derived chemokine (MDC) and intercellular adhesion molecule (ICAM)-1 are known to be involved in allergic skin diseases, such as eczema and atopic dermatitis. Activation of the IFN-γ and its downstream interleukin-12 (IL-12) pathway can correct these diseases. Suppressor of cytokine signaling 1 (SOCS1) is a cytokine signaling inhibitor that blocks downstream pathways of IFN-γ by blocking the mitogen-activated protein kinase (MAPK) and protein kinase B (Akt) signaling pathways. Oxymatrine (OMT), a quinolizidine alkaloid extracted from the herbal medicine Radix Sophorae flavescentis, is used to treat allergic skin diseases in China. The non-cytotoxic concentrations of OMT in HaCaT cells were determined through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Tumor necrosis factor (TNF)-α and IFN-γ were used to stimulate HaCaT cells, and OMT was added to this system with tacrolimus (FK506) as a positive control. The mRNAs of cytokines, MDC, ICAM-1, IL-12p35, IL-12p40, and IFN-γ receptor (IFN-γR)α were detected by RT-PCR. Western blot analyses were performed to assess activation of the MAPK (p38, Jun N-terminal kinase, and extracellular signal-regulated kinase) and Akt signaling pathways. OMT increased the mRNA levels of the IL-12 and IFN-γRα, reduced the mRNA levels of ICAM-1, MDC, and SOCS1. But FK506 increased the mRNA levels of IL12 and inhibited the expression of ICAM-1 mRNAs and had no effects on the IFN-γRα, MDC, and SOCS1 mRNA in HaCaT cells stimulated with TNF-α and IFN-γ. Thus, the mechanisms through which OMT and FK506 ameliorate allergic skin diseases differ.


Assuntos
Alcaloides/farmacologia , Quinolizinas/farmacologia , Dermatopatias/tratamento farmacológico , Linhagem Celular , Quimiocina CCL22 , Regulação para Baixo/efeitos dos fármacos , Humanos , Imunossupressores , Molécula 1 de Adesão Intercelular , Interferon gama/metabolismo , Queratinócitos/citologia , Sistema de Sinalização das MAP Quinases , RNA Mensageiro/efeitos dos fármacos , Dermatopatias/imunologia , Proteína 1 Supressora da Sinalização de Citocina , Tacrolimo/farmacologia
4.
Dalton Trans ; 46(43): 15124-15129, 2017 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-29068016

RESUMO

Two porphyrin chromophores, P1 and P2, were prepared and used as antenna units to coordinate with a metal-free organic dye, JH1, containing pyridine groups. This supramolecular self-assembly strategy can not only effectively improve the light-harvesting ability of the devices but also effectively reduces electron recombination by preventing I3- of the electrolyte from penetrating into the TiO2 surface. The DSSC based on JH1 showed a PCE of 2.46%, with a Voc of 615 mV, Jsc of 6.54 mA cm-2, and FF of 61.18%. After supramolecular self-assembly, the Jsc and Voc of the device were greatly improved. Specifically for the device based on JH1 + P2, the PCE reached 4.39%, which is about 78% greater than the PCE of the device based on JH1; this is mainly due to the Jsc increase of 2.85 mA cm-2 and the Voc increase of 93 mV. Compared to co-sensitization, supramolecular self-assembly does not require tedious optimization steps; thus, this may be a promising and convenient way to improve the overall performance of DSSCs.

5.
PLoS One ; 12(5): e0177600, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28542320

RESUMO

Arrhythmogenesis in acute myocardial infarction (MI) is associated with depolarization of resting membraine potential (RMP) and decrease of inward rectifier potassium current (IK1) in cardiomyocytes. However, clinical anti-arrhythmic agents that primarily act on RMP by enhancing the IK1 channel are not currently available. We hypothesized that zacopride, a selective and moderate agonist of the IK1/Kir2.1 channels, prevents and cures acute ischemic arrhythmias. To test this viewpoint, adult Sprague-Dawley (SD) rats were subjected to MI by ligating the left main coronary artery. The antiarrhythmic effects of zacopride (i.v. infusion) were observed in the settings of pre-treatment (zacopride given 3 min prior to coronary occlusion), post-treatment (zacopride given 3 min after coronary occlusion) and therapeutic treatment (zacopride given 30 s after the onset of the first sustained ventricular tachycardia (VT)/ventricular fibrillation (VF) post MI). In all the three treatment modes, zacopride (15 µg/kg) inhibited MI-induced ventricular tachyarrhythmias, as shown by significant decreases in the premature ventricular contraction (PVC) and the duration and incidence of VT or VF. In Langendorff perfused rat hearts, the antiarrhythmic effect of 1 µmol/L zacopride were reversed by 1 µmol/L BaCl2, a blocker of IK1 channel. Patch clamp results in freshly isolated rat ventricular myocytes indicated that zacopride activated the IK1 channel and thereby reversed hypoxia-induced RMP depolarization and action potential duration (APD) prolongation. In addition, zacopride (1 µmol/L) suppressed hypoxia- or isoproterenol- induced delayed afterdepolarizations (DADs). In Kir2.x transfected Chinese hamster ovary (CHO) cells, zacopride activated the Kir2.1 homomeric channel but not the Kir2.2 or Kir2.3 channels. These results support our hypothesis that moderately enhancing IK1/Kir2.1 currents as by zacopride rescues ischemia- and hypoxia- induced RMP depolarization, and thereby prevents and cures acute ischemic arrhythmias. This study brings a new viewpoint to antiarrhythmic theories and provides a promising target for the treatment of acute ischemic arrhythmias.


Assuntos
Arritmias Cardíacas/tratamento farmacológico , Arritmias Cardíacas/prevenção & controle , Benzamidas/farmacologia , Compostos Bicíclicos Heterocíclicos com Pontes/farmacologia , Isquemia Miocárdica/complicações , Canais de Potássio Corretores do Fluxo de Internalização/agonistas , Potenciais de Ação/efeitos dos fármacos , Doença Aguda , Animais , Arritmias Cardíacas/complicações , Arritmias Cardíacas/patologia , Benzamidas/uso terapêutico , Compostos Bicíclicos Heterocíclicos com Pontes/uso terapêutico , Células CHO , Hipóxia Celular/efeitos dos fármacos , Cricetulus , Isoproterenol/farmacologia , Masculino , Miócitos Cardíacos/efeitos dos fármacos , Miócitos Cardíacos/patologia , Ratos , Ratos Sprague-Dawley
6.
J Agric Food Chem ; 65(20): 4051-4056, 2017 May 24.
Artigo em Inglês | MEDLINE | ID: mdl-28493688

RESUMO

Arctiin is the most abundant bioactive compound contained in the Arctium lappa plant. In our previous study, we isolated one single bacterium capable of bioconverting arctigenin, an aglycone of arctiin, to 3'-desmethylarctigenin (3'-DMAG) solely. However, to date, a specific bacterium capable of producing other arctiin metabolites has not been reported. In this study, we isolated one single bacterium, which we named Eggerthella sp. AUH-JLD49s, capable of bioconverting 3'-DMAG under anaerobic conditions. The metabolite of 3'-DMAG by strain AUH-JLD49s was identified as 3'-desmethyl-4'-dehydroxyarctigenin (DMDH-AG) based on electrospray ionization mass spectrometry (ESI-MS) and 1H and 13C nuclear magnetic resonance spectroscopy. The bioconversion kinetics and bioconversion capacity of strain AUH-JLD49s were investigated. In addition, the metabolite DMDH-AG showed an inhibitory effect on cell growth of human colon cancer cell line HCT116 and human breast cancer cell line MDA-MB-231.


Assuntos
Actinobacteria/isolamento & purificação , Actinobacteria/metabolismo , Furanos/metabolismo , Intestinos/microbiologia , Lignanas/metabolismo , Actinobacteria/genética , Biotransformação , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Fezes/microbiologia , Feminino , Furanos/química , Furanos/farmacologia , Glucosídeos/química , Glucosídeos/metabolismo , Humanos , Lignanas/química , Lignanas/farmacologia , Estrutura Molecular , Espectrometria de Massas por Ionização por Electrospray
7.
Arch Insect Biochem Physiol ; 93(3): 129-142, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27447944

RESUMO

In this study, two full-length cDNA sequences (Cmace1 and Cmace2) encoding putative acetylcholinesterases (AChEs) were cloned and characterized from the rice leaffolder, Cnaphalocrocis medinalis, an important lepidopteran rice pest in Asia. Cmace1 encodes a CmAChE1 consisting of 689 amino acid residues, while Cmace2 encodes a 639 amino acids CmAChE2. The two CmAChEs both have N-terminal signal peptides and conserved motifs including the catalytic triad, choline-binding sites, oxianion hole, acyl pocket, peripheral anionic subsite, and the characteristic FGESAG motif and conserved 14 aromatic amino acids. Phylogenetic analysis showed that Cmace1 and Cmace2 are clustered into distinct clusters that are completely diverged from each other. Reverse-transcription quantitative PCR analysis revealed that Cmace1 and Cmace2 were predominately expressed in the larval brain and at the fifth-instar larvae stage, and the transcription levels of Cmace1 were significantly higher than those of Cmace2 in all the tested samples. Recombinant CmAChE1 and CmAChE2 were heterologously expressed in baculovirus system. Using acetylthiocholine iodide (ATChI) as substrate, the Michaelis constant (Km ) values of rCmAChE1 and rCmAChE2 were 39.81 ± 6.49 and 68.29 ± 6.72 µmol/l, respectively; and the maximum velocity (Vmax ) values of the two rCmAChEs were 0.60 ± 0.02 and 0.31 ± 0.06 µmol/min/mg protein, respectively. Inhibition assay indicated that rCmAChE1 was more sensitive to the organophosphate insecticides chlorpyrifos and triazophos than rCmAChE2. This study is the first report of molecular cloning and biochemical characterization of two acetylcholinesterase genes/enzymes in C. medinalis.


Assuntos
Acetilcolinesterase/genética , Proteínas de Insetos/genética , Mariposas/enzimologia , Mariposas/genética , Acetilcolinesterase/química , Acetilcolinesterase/metabolismo , Sequência de Aminoácidos , Animais , Clonagem Molecular , DNA Complementar/genética , DNA Complementar/metabolismo , Feminino , Proteínas de Insetos/química , Proteínas de Insetos/metabolismo , Larva/genética , Larva/crescimento & desenvolvimento , Larva/metabolismo , Masculino , Mariposas/classificação , Mariposas/crescimento & desenvolvimento , Filogenia , Pupa/genética , Pupa/crescimento & desenvolvimento , Pupa/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Alinhamento de Sequência
8.
Asian Pac J Trop Med ; 9(5): 489-93, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-27261860

RESUMO

OBJECTIVE: To observe the influence of different concentrations of homocysteine (Hcy) and hydrogen sulfide (H2S) on the secretion and activation of matrix metalloproteinase-2 (MMP-2) in cardiocytes so as to search for new ways to fight against myocardial tissue fibrosis. METHODS: Cardiocytes H9C2 was cultured in vitro and different concentrations of Hcy and H2S were added for 6-h and 24-h cultivation. MTT cell proliferation assay was applied to test the activation change of cardiocytes H9C2 after affecting by different concentrations of Hcy and H2S. ELISA and MTT were employed to detect the expression and enzymatic activity of MMP-2. RESULTS: The H9C2 cell inhibition of activity was more significant with 1000 µmol/L of Hcy as compared with other concentrations (P < 0.001). With 2.5-100.0 µmol/L Hcy and 0.1, 1.0 and 10.0 mmol/L H2S, the activity of H9C2 did not change significantly (P > 0.05). Hcy with concentrations of 10, 50 and 100 µmol/L could increase the quantity of MMP-2 secreted by cardiocytes H9C2, and the interaction strength was concentration-dependent (P < 0.05). After interacting with 100 µmol/L of Hcy for 6 h, the zymogen activation effect of MMP-2 was stronger than that of the 2.5-25 µmol/L group (P < 0.05). After interacting with Hcy and H2S (1.0 mmol/L) for 6 h and 24 h, the activation effect of MMP-2 was stronger than those interacted with 10, 25, 50 and 100 µmol/L of Hcy (P < 0.05). CONCLUSIONS: Hcy can increase the production of MMP-2 secreted by H9C2 cell and improve its zymogen activation. Besides, the interaction strength is concentration-dependent; while H2S can up-regulate the activation of MMP-2 and co-promote the activation of MMP-2 with Hcy as well.

9.
Mol Med Rep ; 13(1): 315-20, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26548607

RESUMO

Ubiquitin­like with plant homeodomain (PHD) and RING­finger domain 1 (UHRF1) maintains methylation patterns following DNA replication and is expressed at high levels in various types of human cancer. UHRF1 has been identified as a novel oncogene involved in the pathogenesis of hepatocellular carcinoma. Previous studies have demonstrated that inhibition of the expression of UHRF1 suppresses the proliferation of cancer cells. However, the role of UHRF1 in human osteosarcoma has not been investigated. The present study examined the expression levels of UHRF1 and retinoblastoma 1 (Rb1) in human osteosarcoma cell lines by western blot analysis. Stable overexpression of UHRF1 or knockdown of Rb1 was achieved by lentiviral transfection. Subsequently, a Cell Counting Kit-8 assay and a cell invasion assay were performed to detect the biological functions of UHRF1 in vitro. The results of the present study demonstrated that UHRF1 promoted the proliferation of human osteosarcoma cells. The present study also reported that UHRF1 was able to enhance the invasion of osteosarcoma cells in a retinoblastoma 1 (Rb1)­dependent manner. UHRF1 promoted invasion in Rb1­positive osteosarcoma cells, but not in Saos­2 cells with homozygous loss of Rb1. Similarly, knockdown of Rb1 in Rb1­positive osteosarcoma cells enhanced levels of invasion and eliminated the regulation of invasion by UHRF1. UHRF1 was found to inhibit the mRNA and protein expression levels of Rb1. Furthermore, deletion of Rb1 was found to suppress the expression of E­cadherin and promote epithelial­to­mesenchymal transition (EMT). In addition, the overexpression of UHRF1 inhibited the expression of E­cadherin and promoted EMT via the suppression of Rb1. These data demonstrated that UHRF1 promotes osteosarcoma cell invasion by downregulating the expression of E­cadherin and increasing EMT in an Rb1­dependent manner.


Assuntos
Proteínas Estimuladoras de Ligação a CCAAT/metabolismo , Caderinas/metabolismo , Regulação para Baixo , Osteossarcoma/genética , Osteossarcoma/patologia , Proteína do Retinoblastoma/metabolismo , Linhagem Celular Tumoral , Proliferação de Células , Transição Epitelial-Mesenquimal/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Invasividade Neoplásica , Proteína do Retinoblastoma/genética , Ubiquitina-Proteína Ligases
10.
Curr Genomics ; 16(6): 393-404, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27019614

RESUMO

Osteoarthritis (OA) is an age-related disease with poorly understood pathogenesis. Recent studies have demonstrated that miRNA might play a key role in OA initiation and development. We reviewed recent publications and elucidated the connection between miRNA and OA cartilage anabolic and catabolic signals, including four signaling pathways: TGF-ß/Smads and BMPs signaling, associated with cartilage anabolism; and MAPK and NF-KB signaling, associated with cartilage catabolism. We also explored the relationships with MMP, ADAMTS and NOS (NitricOxide Synthases) families, as well as with the catabolic cytokines IL-1 and TNF-α. The potential role of miRNAs in biological processes such as cartilage degeneration, chondrocyte proliferation, and differentiation is discussed. Collective evidence indicates that miRNAs play a critical role in cartilage degeneration. These findings will aid in understanding the molecular network that governs articular cartilage homeostasis and in to elucidate the role of miRNA in the pathogenesis of OA.

11.
Zhonghua Yi Xue Za Zhi ; 92(13): 899-903, 2012 Apr 03.
Artigo em Chinês | MEDLINE | ID: mdl-22781531

RESUMO

OBJECTIVE: To localize the sensory motor cortex of human brain by analyzing the power change in Gamma band (> 60 Hz) of electrocorticography (ECoG) data. METHODS: Eight patients with intractable epilepsy underwent temporary placement of subdural electrodes. After surgery, sensory evoked potential (SEP), electrocortical stimulation (CES) and event-related synchronization analysis of Gamma band (Gamma ERS analysis) were performed to reduce the risk of complications. The results of Gamma ERS analysis were compared with those of SEP and CES. RESULTS: The results of Gamma ERS analysis had 80.7% electrodes fitting perfectly those of CES and SEP. And the percentage reached 92.3% if electrodes were superimposed or added adjacently. CONCLUSION: The Gamma ERS analysis is a new sensitive and precise method for cortical function mapping.


Assuntos
Eletroencefalografia , Epilepsia/fisiopatologia , Córtex Motor/fisiopatologia , Adolescente , Adulto , Mapeamento Encefálico , Estimulação Elétrica , Eletrodos , Potenciais Evocados , Feminino , Humanos , Masculino , Adulto Jovem
12.
J Microbiol Biotechnol ; 22(3): 343-51, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22450790

RESUMO

In this paper, the kinetics of a cloned special glucosidase, named ginsenosidase type III hydrolyzing 3-O-glucoside of multi-protopanaxadiol (PPD)-type ginsenosides, were investigated. The gene (bgpA) encoding this enzyme was cloned from a Terrabacter ginsenosidimutans strain and then expressed in E. coli cells. Ginsenosidase type III was able to hydrolyze 3-O-glucoside of multi-PPD-type ginsenosides. For instance, it was able to hydrolyze the 3-O-ß-D-(1-->2)-glucopyranosyl of Rb1 to gypenoside XVII, and then to further hydrolyze the 3-O-ß-D-glucopyranosyl of gypenoside XVII to gypenoside LXXV. Similarly, the enzyme could hydrolyze the glucopyranosyls linked to the 3-O- position of Rb2, Rc, Rd, Rb3, and Rg3. With a larger enzyme reaction Km value, there was a slower enzyme reaction speed; and the larger the enzyme reaction Vmax value, the faster the enzyme reaction speed was. The Km values from small to large were 3.85 mM for Rc, 4.08 mM for Rb1, 8.85 mM for Rb3, 9.09 mM for Rb2, 9.70 mM for Rg3(S), 11.4 mM for Rd and 12.9 mM for F2; and Vmax value from large to small was 23.2 mM/h for Rc, 16.6 mM/h for Rb1, 14.6 mM/h for Rb3, 14.3 mM/h for Rb2, 1.81mM/h for Rg3(S), 1.40 mM/h for Rd, and 0.41 mM/h for F2. According to the Vmax and Km values of the ginsenosidase type III, the hydrolysis speed of these substrates by the enzyme was Rc>Rb1>Rb3>Rb2>Rg3(S)>Rd>F2 in order.


Assuntos
Actinomycetales/enzimologia , Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Ginsenosídeos/metabolismo , Glucosidases/química , Glucosidases/metabolismo , Sapogeninas/metabolismo , Actinomycetales/genética , Proteínas de Bactérias/genética , Clonagem Molecular , Ginsenosídeos/química , Glucosidases/genética , Cinética , Estrutura Molecular , Sapogeninas/química , Especificidade por Substrato
13.
J Microbiol Biotechnol ; 21(10): 1057-63, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22031031

RESUMO

Herein, a novel ginsenosidase, named ginsenosidase type IV, hydrolyzing 6-O-multi-glycosides of protopanaxatrioltype ginsenosides (PPT), such as Re, R1, Rf, and Rg2, was isolated from the Aspergillus sp. 39g strain, purified, and characterized. Ginsenosidase type IV was able to hydrolyze the 6-O-alpha-L-(1-->2)-rhamnoside of Re and the 6-O-beta-D- (1-->2)-xyloside of R1 into ginsenoside Rg1. Subsequently, it could hydrolyze the 6-O-beta-D-glucoside of Rg1 into F1. Similarly, it was able to hydrolyze the 6-O-alpha-L-(1-->2)- rhamnoside of Rg2 and the 6-O-beta-D-(1-->2)-glucoside of Rf into Rh1, and then further hydrolyze Rh1 into its aglycone. However, ginsenosidase type IV could not hydrolyze the 3-O- or 20-O-glycosides of protopanaxadioltype ginsenosides (PPD), such as Rb1, Rb2, Rb3, Rc, and Rd. These exhibited properties are significantly different from those of glycosidases described in Enzyme Nomenclature by the NC-IUBMB. The optimal temperature and pH for ginsenosidase type IV were 40°C and 6.0, respectively. The activity of ginsenosidase type IV was slightly improved by the Mg(2+) ion, and inhibited by Cu(2+) and Fe(2+) ions. The molecular mass of the enzyme, based on SDS-PAGE, was noted as being approximately 56 kDa.


Assuntos
Aspergillus/enzimologia , Proteínas Fúngicas/metabolismo , Ginsenosídeos/metabolismo , Glicosídeo Hidrolases/metabolismo , Aspergillus/química , Aspergillus/genética , Estabilidade Enzimática , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Proteínas Fúngicas/isolamento & purificação , Ginsenosídeos/química , Glicosídeo Hidrolases/química , Glicosídeo Hidrolases/genética , Glicosídeo Hidrolases/isolamento & purificação , Hidrólise , Especificidade por Substrato
14.
Blood Coagul Fibrinolysis ; 21(5): 420-4, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20442653

RESUMO

The mechanism of testosterone inducing the tissue factor pathway inhibitor (TFPI) in protecting against thrombosis is unknown. We aimed to elucidate the mechanisms involved in the induction by observing, in human umbilical vein endothelial cells (HUVECs), the phosphorylation of mitogen-activated protein kinases (MAPKs), a major cell signaling system. The level of testosterone regulating several signaling pathways, including extracellular signal-regulated kinases 1/2 (ERK1/2), c-Jun-N-terminal kinase (JNK), and p38 MAPK, was measured by western blot in HUVECs. ELISA and quantitative real-time reverse transcriptase-PCR were used to analyze TFPI expression after blocking ERK1/2 (with PD98059) or JNK (with SP600125) pathway in HUVECs. Testosterone-induced a rapid phosphorylation of ERK1/2, JNK and p38 MAPK in HUVECs, which could not be inhibited by androgen receptor antagonist flutamide. Blocking ERK1/2 or JNK pathway could significantly impair testosterone-induced TFPI at both translational and transcriptional levels in HUVECs. Testosterone at a physiological concentration may help to prevent thrombosis development by stimulating TFPI expression in HUVECs, partly through the ERK1/2 and JNK MAPK pathway.


Assuntos
Células Endoteliais/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Lipoproteínas/biossíntese , Lipoproteínas/genética , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Testosterona/farmacologia , Células Cultivadas , Células Endoteliais/metabolismo , Ensaio de Imunoadsorção Enzimática , Humanos , Lipoproteínas/metabolismo , Proteínas Quinases Ativadas por Mitógeno/antagonistas & inibidores , Fosforilação/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Trombose/prevenção & controle
15.
Clin Chem Lab Med ; 47(3): 327-33, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-19676145

RESUMO

BACKGROUND: The increased expression of heme oxygenase-1 content, a stress-response protein, directly correlates with the incidence of coronary heart disease. Down-regulation of hypoxia inducible factor-1alpha activity, a major downstream effector of the signaling pathways activated by hypoxia, increases cell survival after hypoxia. The ubiquitin system, a non-lysosomal pathway of protein degradation, is involved in processes of coronary heart disease. The aim of this study was to investigate the expression of heme oxygenase-1, hypoxia inducible factor-1alpha, and ubiquitin in both monocytes and lymphocytes isolated from patients at the mRNA and protein levels in different stages of coronary heart disease and their possible correlation. METHODS: A total of 90 patients with coronary heart disease (30 acute myocardial infarction, 30 unstable angina pectoris, and 30 stable angina pectoris) were selected, and 30 cases with normal coronary artery served as controls. The mRNA and protein expression of heme oxygenase-1, hypoxia inducible factor-1alpha, and ubiquitin in monocytes and lymphocytes were examined by semi-quantitative reverse transcriptase polymerase chain reaction and Western blotting, respectively. RESULTS: The mRNA expression of heme oxygenase-1 and ubiquitin was associated with the severity of coronary heart disease (p<0.05). There was no significant difference in hypoxia inducible factor-1alpha mRNA expression between the coronary heart disease patients and controls. The protein expression of heme oxygenase-1, hypoxia inducible factor-1alpha, and ubiquitin was significantly stronger in patients with coronary heart disease than in controls, and the expression levels increased with the severity of the disease. There was a positive association between heme oxygenase-1 and hypoxia inducible factor-1alpha and ubiquitin, antioxidative therapy with adrenergic receptor blocker, angiotensin-converting enzyme inhibitor or statins up-regulated the expression of heme oxygenase-1 and hypoxia inducible factor-1alpha. CONCLUSIONS: These data suggest that heme oxygenase-1, hypoxia inducible factor-1alpha, and ubiquitin are involved in the development and progression of coronary heart disease and thus may be useful biomarkers for coronary heart disease.


Assuntos
Doença das Coronárias/genética , Perfilação da Expressão Gênica , Heme Oxigenase-1/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Leucócitos Mononucleares/imunologia , Linfócitos/imunologia , Ubiquitina/genética , Bloqueadores do Receptor Tipo 1 de Angiotensina II/farmacologia , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Doença das Coronárias/imunologia , Feminino , Heme Oxigenase-1/efeitos dos fármacos , Heme Oxigenase-1/metabolismo , Humanos , Inibidores de Hidroximetilglutaril-CoA Redutases/farmacologia , Subunidade alfa do Fator 1 Induzível por Hipóxia/efeitos dos fármacos , Inflamação , Masculino , Pessoa de Meia-Idade , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Ubiquitina/efeitos dos fármacos
16.
Asian J Androl ; 11(2): 266-71, 2009 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-19169266

RESUMO

We have observed earlier that testosterone at physiological concentrations can stimulate tissue factor pathway inhibitor (TFPI) gene expression through the androgen receptor in endothelial cells. This study further investigated the impact of testosterone on TFPI levels in response to inflammatory cytokine tumor necrosis factor-alpha (TNF-alpha). Cultured human umbilical vein endothelial cells were incubated in the presence or absence of testosterone or TNF-alpha. TFPI protein and mRNA levels were assessed by enzyme-linked immunosorbent assay and quantitative real-time reverse transcription polymerase chain reaction. To study the cellular mechanism of testosterone's action, nuclear factor-kappa B (NF-kappaB) translocation was confirmed by electrophoretic mobility shift assays. We found that after NF-kappaB was activated by TNF-alpha, TFPI protein levels declined significantly by 37.3% compared with controls (P < 0.001), and the mRNA levels of TFPI also decreased greatly (P < 0.001). A concentration of 30 nmol L(-1) testosterone increased the secretion of TFPI compared with the TNF-alpha-treated group. NF-kappaB DNA-binding activity was significantly suppressed by testosterone (P < 0.05). This suggests that physiological testosterone concentrations may exert their antithrombotic effects on TFPI expression during inflammation by downregulating NF-kappaB activity.


Assuntos
Androgênios/farmacologia , Regulação para Baixo/efeitos dos fármacos , Endotélio Vascular/efeitos dos fármacos , Lipoproteínas/genética , Subunidade p50 de NF-kappa B/antagonistas & inibidores , Testosterona/farmacologia , Fator de Necrose Tumoral alfa/farmacologia , Células Cultivadas , Combinação de Medicamentos , Endotélio Vascular/metabolismo , Humanos , Recém-Nascido , Lipoproteínas/metabolismo , Subunidade p50 de NF-kappa B/genética , RNA Mensageiro/metabolismo
17.
Coron Artery Dis ; 19(3): 173-9, 2008 May.
Artigo em Inglês | MEDLINE | ID: mdl-18418234

RESUMO

BACKGROUND: Marked variability exists in coronary artery collaterals in patients with ischemic heart disease. Multiple factors are thought to play a role in collateral development; however, the contribution of hypoxia inducible factor-1alpha (HIF-1alpha), which is a transcriptional activator that functions as a master regulator of oxygen homeostasis, is not completely clear. It could play an important role in modulating collateral development. OBJECTIVE: The objective of this study is to investigate the changes and significance of expression of HIF-1alpha in patients with coronary artery collaterals. METHODS: Collateral vessels were determined in 98 patients with >or=70% narrowing of at least one coronary artery without earlier revascularization, 42 patients with coronary artery collaterals and 56 patients with no coronary artery collaterals. Extent of collaterals was expressed as scores according to the Rentrop scoring system. Another 50 cases with normal coronary arteries were selected as control. The levels of HIF-1alpha protein expression in monocyte and lymphocyte in the participants were tested by immunohistochemistry (IHC) and western blot; mRNA levels were measured using reverse transcriptase PCR technique. RESULTS: Compared with the control with normal coronary artery, the patients had higher expression of HIF-1alpha protein tested by IHC and western blot (52.6+/-10.2 vs. 13.7+/-6.2 by IHC, 50.8+/-4.5 vs. 6.5+/-1.8 by western blot); furthermore, significantly higher HIF-1alpha expression was observed in patients with collaterals compared with patients with no collaterals (81.5+/-11.8 vs. 20.7+/-9.4 by IHC; 87.2+/-6.5 vs. 9.5+/-1.4 by western blot). On the transcriptional levels of HIF-1alpha, the result was the same as the protein, there was significant difference of HIF-1alpha between the three groups. The patients with collaterals were the highest (127.3+/-23.9), followed by patients with no collaterals (35.7+/-12.3), and the control were the lowest (23.5+/-9.3). A highly positive correlation was observed between the expression/transcription of HIF-1alpha and collateral score (P<0.01, IHC: r1=0.78, reverse transcriptase PCR: r2=0.69, western blot: r3=0.84). CONCLUSION: These data suggest that higher inductions of HIF-1alpha are associated with coronary collaterals, thus implying that HIF-1alpha may promote coronary collateral formation. Detection of HIF-1alpha expression might be helpful to predict prognosis of patients with coronary artery disease.


Assuntos
Circulação Colateral , Doença da Artéria Coronariana/sangue , Subunidade alfa do Fator 1 Induzível por Hipóxia/sangue , Idoso , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , RNA Mensageiro/análise
18.
Zhonghua Nan Ke Xue ; 13(9): 777-9, 2007 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-17929550

RESUMO

OBJECTIVE: To investigate the effects of testosterone on extracellular signal-regulated kinase l/2 ( ERK1/2) phosphorylation in human umbilical vein endothelial cells (HUVEC). METHODS: Activations of ERK1/2 stimulated by physiological testosterone were detected by Western blotting in cultured HUVEC. RESULTS: A rapid phosphorylation expression of ERK1/2 was observed by treatment of the HUVECs with 3 x 10(-8) mol/L testosterone, especially at 30 minutes. This phosphorylation was greatly inhibited by incubation with androgen receptor antagonist flutamide for 3 hours previously. CONCLUSION: Testosterone at physiological concentrations induces the mitogen-activated protein kinase (MAPK, ERK1/2 and MEK1/2) phosphorylation within a short time, and flutamide could impair the process.


Assuntos
Células Endoteliais/efeitos dos fármacos , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Testosterona/farmacologia , Antagonistas de Receptores de Andrógenos , Western Blotting , Células Cultivadas , Células Endoteliais/citologia , Células Endoteliais/metabolismo , Ativação Enzimática/efeitos dos fármacos , Humanos , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Fosforilação/efeitos dos fármacos , Receptores Androgênicos/metabolismo , Veias Umbilicais/citologia
19.
Biochem Cell Biol ; 85(2): 246-51, 2007 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-17534406

RESUMO

There is a striking gender difference in atherosclerotic vascular disease. For decades, testosterone was considered detrimental to the cardiovascular system. Recent studies, however, have presented some alternative results. The aim of this study was to evaluate the effect of testosterone, using physiological and supraphysiological concentrations, on antigen and mRNA levels of tissue plasminogen activator (tPA), plasminogen activator inhibitor type 1 (PAI-1), and tissue factor pathway inhibitor (TFPI) released by human umbilical vein endothelial cells and to investigate the cellular mechanism. Cells within 2-3 passages were cultured in 25 cm(2) flasks or plated onto 96-well plates with a density of about 1 x 10(5) cells/mL as recommended. The cells were incubated in the presence or absence of testosterone (3, 30, 3 x 10(3), 3 x 10(4) nmol/L) for 48 h. Levels of tPA, PAI-1, and TFPI antigen were assayed with ELISA kits. Reverse transcriptase PCR was carried out to detect tPA, PAI-1, and TFPI mRNA levels. Cells were incubated in androgen-receptor antagonist (flutamide 10 micromol/L) or aromatase inhibitor (aminoglutethimide 50 micromol/L) for 3 h, and then the experiments were repeated. Testosterone at a physiologic concentration (30 nmol/L) increased the antigen levels of tPA and TFPI significantly (P < 0.05). However, tPA and TFPI levels were markedly reduced (P < 0.05) at a larger dose (3 x 10(4) nmol/L). On the other hand, PAI-1 antigen levels decreased significantly at the testosterone concentrations ranging from 3 to 3 x 10(4) nmol/L (P < 0.05). The change in the levels of tPA and TFPI were reflected in the corresponding change in mRNA levels. Flutamide attenuated the effect of testosterone at physiological concentration (30 nmol/L). The results demonstrated that testosterone at physiological concentrations may have a beneficial influence on the haemostatic system through enhancement of anticoagulant activity, resulting from stimulation of TFPI and tPA expression and inhibition of PAI-1 secretion by the endothelium.


Assuntos
Androgênios/farmacologia , Células Endoteliais/metabolismo , Lipoproteínas/biossíntese , Inibidor 1 de Ativador de Plasminogênio/biossíntese , Ativadores de Plasminogênio/biossíntese , Testosterona/farmacologia , Antagonistas de Androgênios/farmacologia , Células Cultivadas , Relação Dose-Resposta a Droga , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/fisiologia , Células Endoteliais/citologia , Flutamida/farmacologia , Humanos , Recém-Nascido , Lipoproteínas/genética , Masculino , Inibidor 1 de Ativador de Plasminogênio/genética , Ativadores de Plasminogênio/genética , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Testosterona/metabolismo , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/fisiologia
20.
Acta Pharmacol Sin ; 28(1): 36-43, 2007 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-17184580

RESUMO

AIM: To study whether urotensin II (UII), a potent vasoconstrictive peptide, is involved in the development of cardiac hypertrophy and fibrogenesis of rats induced by isoproterenol (ISO). METHODS: Thirty male Wistar rats were randomly divided into 3 groups. Group 1 was the healthy control group, group 2 was the ISO group, and group 3 was the ISO+UII group. In groups 2 and 3, ISO (5 mg x kg(-1) x d(-1)) was given (sc) once daily for 7 d. Group 3 was also given UII in the first day [3 nmol/kg (5 microg/kg), iv], followed by sc (1.5 microg/kg) twice daily. Group 1 received 0.9% saline. UII receptor (UT) mRNA expression was determined by RT-PCR. The contents of UII and angiotensin II (Ang II) were determined by radioimmunoassay. In vitro, the effects of UII on DNA/collagen synthesis of cardiac fibroblasts were determined by [3H]thymidine/[3H]proline incorporation. RESULTS: The ratio of heart weight/body weight, plasma lactate dehydrogenase activity, myocardial malondialdehyde and hydroxyproline concentration increased significantly in the ISO group, as well as UT mRNA expression, plasma and cardiac UII and ventricular Ang II, compared with the control group (P< 0.01). ISO induced significant myocardial fibrogenesis. Moreover, UII+ISO co-treatment significantly increased the changes of biochemical markers of injury and the degree of cardiac hypertrophy and fibrosis. In vitro, 5 x 10(-9 )-5 x 10(-7 ) mol/L UII stimulated [3H]thymidine/[3H] proline incorporation into cardiac fibroblasts in a dose-dependent manner (P< 0.01). CONCLUSION: These results suggest that UII was involved in the development of cardiac fibrosis and hypertrophy by synergistic effects with ISO.


Assuntos
Cardiomegalia/metabolismo , Miocárdio/metabolismo , Urotensinas/farmacologia , Angiotensina II/metabolismo , Animais , Cardiomegalia/sangue , Cardiomegalia/induzido quimicamente , Proliferação de Células/efeitos dos fármacos , Colágeno/metabolismo , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Fibrose , Hidroxiprolina/metabolismo , Isoproterenol , Lactato Desidrogenases/sangue , Masculino , Malondialdeído/metabolismo , Miocárdio/patologia , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Distribuição Aleatória , Ratos , Ratos Wistar , Receptores Acoplados a Proteínas G/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Urotensinas/metabolismo
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