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1.
Rev. bras. ginecol. obstet ; 41(7): 449-453, July 2019. tab
Article in English | LILACS | ID: biblio-1020606

ABSTRACT

Abstract Objective To analyze the effects of estrogen alone or in combination with progestogens and tibolone (TIB) on the expression of the extracellular matrix metalloproteinases 2 and 9 (MMP-2 and MMP-9), of perlecan, and of heparanase (HPSE) of the vascular walls of the carotid arteries. Methods A total of 30 250-day-old ovariectomized Wistar rats were orally treated for 5 weeks with: a) 1 mg/kg of estradiol benzoate (EB); b) EB + 0.2 mg/kg of medroxyprogesterone acetate (MPA); c) EB + 0.2mg/kg of norethisterone acetate (NETA); d) EB + 2 mg/kg of dydrogesterone (DI); e) 1 mg/kg of TIB; f) placebo (CTR). Following treatment, the expression of mRNA for MMP-2, MMP-9, and HPSE was analyzed by realtime polymerase chain-reaction (PCR), and the expression of MMP-2, of MMP-9, of tissue inhibitor of metalloproteinase 2 (TIMP-2), and of perlecan was quantified by immunohistochemistry in the carotid arteries. Results The groups showed significant differences on mRNA HPSE expression (p = 0.048), which was higher in the EB, EB + MPA, and TIB groups. There was no statistically significant difference in mRNA MMP-2 or MMP-9 expression. The immunohistochemical expression of MMP-2, of TIMP-2, of MMP-9, of HPSE, and of perlecan showed no differences between groups. Conclusion Estradiol alone or associated with MPA and TIB treatment can increase mRNA HSPE expression of the walls of the carotid arteries in ovariectomized rats.


Resumo Objetivo Analisar os efeitos do estrogênio isolado ou em combinação com progestogênios e tibolona (TIB) na expressão das metaloproteinases 2 e 9 da matriz extracelular (MMP-2 e MMP-9), da perlecan e da heparanase (HPSE) das paredes vasculares das artérias carótidas. Métodos Trinta ratas Wistar ovariectomizadas com 250 dias de idade foram tratadas oralmente por 5 semanas com: a) 1 mg/kg de benzoato de estradiol (EB); b) EB + 0,2 mg/kg de acetato de medroxiprogesterona (MPA); c) EB + 0,2mg/kg de acetato de noretisterona (NETA); d) EB + 2 mg/kg de didrogesterona (DI); e) 1 mg/kg de TIB; f) placebo (CTR). Após o tratamento, a expressão de mRNA para MMP-2, MMP- 9, e HPSE foi analisada por reação em cadeia da polimerase (RCP) em tempo real, e a expressão de MMP-2, MMP-9, inibidor tecidual de metaloproteinase 2 (TIMP-2), e de perlecan foi quantificado por imunohistoquímica em artérias carótidas. Resultados Os grupos apresentaram diferenças significativas na expressão do mRNA HPSE (p = 0,048), sendo maiores nos grupos EB, EB + MPA e TIB. Não houve diferença estatisticamente significativa nas expressões de mRNA MMP-2 ou MMP-9. A expressão imunohistoquímica de MMP-2, TIMP-2, MMP-9, HPSE e perlecan não mostrou diferenças entre os grupos. Conclusão O estradiol isolado ou associado ao tratamento com MPA e TIB pode aumentar a expressão de mRNA HSPE nas paredes das artérias carótidas em ratas ovariectomizadas.


Subject(s)
Animals , Female , Rats , Progestins/pharmacology , Carotid Arteries/enzymology , Heparin Lyase/drug effects , Estradiol/analogs & derivatives , Contraceptive Agents, Hormonal/pharmacology , Norpregnenes/pharmacology , Progestins/administration & dosage , Ovariectomy , Carotid Arteries/drug effects , Estrogen Replacement Therapy , Gene Expression Regulation, Enzymologic/drug effects , Administration, Oral , Rats, Wistar , Heparin Lyase/genetics , Heparin Lyase/metabolism , Heparan Sulfate Proteoglycans/genetics , Heparan Sulfate Proteoglycans/metabolism , Matrix Metalloproteinase 2/drug effects , Matrix Metalloproteinase 2/genetics , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/drug effects , Matrix Metalloproteinase 9/genetics , Matrix Metalloproteinase 9/metabolism , Models, Animal , Estradiol/administration & dosage , Estradiol/pharmacology , Contraceptive Agents, Hormonal/administration & dosage , Norpregnenes/administration & dosage
2.
Braz. j. med. biol. res ; 51(4): e6980, 2018. graf
Article in English | LILACS | ID: biblio-889067

ABSTRACT

Hormones regulate hepatic gene expressions to maintain metabolic homeostasis. Ectonucleotide pyrophosphatase/phosphodiesterase 1 has been thought to interfere with insulin signaling. To determine its potential role in the regulation of metabolism, we analyzed its gene (Enpp1) expression in the liver of rats experiencing fasting and refeeding cycles, and in primary rat hepatocytes and human hepatoma HepG2 cells treated with insulin and dexamethasone using northern blot and real-time PCR techniques. Hepatic Enpp1 expression was induced by fasting and reduced by refeeding in the rat liver. In primary rat hepatocytes and HepG2 hepatoma cells, insulin reduced Enpp1 mRNA abundance, whereas dexamethasone induced it. Dexamethasone disrupted the insulin-reduced Enpp1 expression in primary hepatocytes. This is in contrast to the responses of the expression of the cytosolic form of phosphoenolpyruvate carboxykinase gene to the same hormones, where insulin reduced it significantly in the process. In addition, the dexamethasone-induced Enpp1 gene expression was attenuated in the presence of 8-Br-cAMP. In conclusion, we demonstrated for the first time that hepatic Enpp1 is regulated in the cycle of fasting and refeeding, a process that might be attributed to insulin-reduced Enpp1 expression. This insulin-reduced Enpp1 expression might play a role in the development of complications in diabetic patients.


Subject(s)
Humans , Animals , Male , Rats , Pyrophosphatases/genetics , RNA, Messenger/drug effects , Dexamethasone/pharmacology , Phosphoric Diester Hydrolases/genetics , Glucocorticoids/pharmacology , Hypoglycemic Agents/pharmacology , Insulin/pharmacology , Liver/enzymology , Pyrophosphatases/biosynthesis , Pyrophosphatases/drug effects , Insulin Resistance , RNA, Messenger/metabolism , Gene Expression Regulation, Enzymologic/drug effects , Enzyme Induction/drug effects , Fasting/metabolism , Rats, Sprague-Dawley , Phosphoric Diester Hydrolases/biosynthesis , Phosphoric Diester Hydrolases/drug effects , Hep G2 Cells , Real-Time Polymerase Chain Reaction
3.
Braz. j. microbiol ; 47(3): 691-696, July-Sept. 2016. tab
Article in English | LILACS | ID: lil-788958

ABSTRACT

ABSTRACT This study highlights the prevalence of aminoglycoside-modifying enzyme genes and virulence determinants among clinical enterococci with high-level aminoglycoside resistance in Inner Mongolia, China. Screening for high-level aminoglycoside resistance against 117 enterococcal clinical isolates was performed using the agar-screening method. Out of the 117 enterococcal isolates, 46 were selected for further detection and determination of the distribution of aminoglycoside-modifying enzyme-encoding genes and virulence determinants using polymerase chain reaction -based methods. Enterococcus faecium and Enterococcus faecalis were identified as the species of greatest clinical importance. The aac(6')-Ie-aph(2")-Ia and ant(6')-Ia genes were found to be the most common aminoglycoside-modifying enzyme genes among high-level gentamicin resistance and high-level streptomycin resistance isolates, respectively. Moreover, gelE was the most common virulence gene among high-level aminoglycoside resistance isolates. Compared to Enterococcus faecium, Enterococcus faecalis harbored multiple virulence determinants. The results further indicated no correlation between aminoglycoside-modifying enzyme gene profiles and the distribution of virulence genes among the enterococcal isolates with high-level gentamicin resistance or high-level streptomycin resistance evaluated in our study.


Subject(s)
Male , Female , Gene Expression Regulation, Bacterial/drug effects , Gene Expression Regulation, Enzymologic/drug effects , Enterococcus/drug effects , Enterococcus/physiology , Drug Resistance, Bacterial , Aminoglycosides/metabolism , Aminoglycosides/pharmacology , Virulence/genetics , Microbial Sensitivity Tests , China/epidemiology , Prevalence , Gram-Positive Bacterial Infections/microbiology , Gram-Positive Bacterial Infections/epidemiology , Enterococcus/metabolism , Genes, Bacterial , Anti-Bacterial Agents/metabolism
4.
Braz. j. med. biol. res ; 48(7): 654-664, 07/2015. graf
Article in English | LILACS | ID: lil-751344

ABSTRACT

Recent evidence indicates that a deficiency of 1,25-dihydroxyvitamin D3 (1,25[OH]2D3) may influence asthma pathogenesis; however, its roles in regulating specific molecular transcription mechanisms remain unclear. We aimed to investigate the effect of 1,25(OH)2D3 on the expression and enzyme activity of histone deacetylase 2 (HDAC2) and its synergistic effects with dexamethasone (Dx) in the inhibition of inflammatory cytokine secretion in a rat asthma model. Healthy Wistar rats were randomly divided into 6 groups: control, asthma, 1,25(OH)2D3 pretreatment, 1,25(OH)2D3 treatment, Dx treatment, and Dx and 1,25(OH)2D3 treatment. Pulmonary inflammation was induced by ovalbumin (OVA) sensitization and challenge (OVA/OVA). Inflammatory cells and cytokines in the bronchoalveolar lavage (BAL) fluid and histological changes in lung tissue were examined. Nuclear factor kappa B (NF-κB) p65 and HDAC2 expression levels were assessed with Western blot analyses and quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR). Enzyme activity measurements and immunohistochemical detection of HDAC2 were also performed. Our data demonstrated that 1,25(OH)2D3 reduced the airway inflammatory response and the level of inflammatory cytokines in BAL. Although NF-κB p65 expression was attenuated in the pretreatment and treatment groups, the expression and enzyme activity of HDAC2 were increased. In addition, 1,25(OH)2D3 and Dx had synergistic effects on the suppression of total cell infusion, cytokine release, and NF-κB p65 expression, and they also increased HDAC2 expression and activity in OVA/OVA rats. Collectively, our results indicated that 1,25(OH)2D3 might be useful as a novel HDAC2 activator in the treatment of asthma.


Subject(s)
Animals , Male , Asthma/drug therapy , Calcitriol/pharmacology , /drug effects , NF-kappa B/drug effects , Vitamins/pharmacology , Asthma/chemically induced , Blotting, Western , Bronchoalveolar Lavage Fluid/chemistry , Cell Count , Calcitriol/therapeutic use , Cytokines/analysis , Cytokines/drug effects , Disease Models, Animal , Enzyme-Linked Immunosorbent Assay , Gene Expression Regulation, Enzymologic/drug effects , /metabolism , Immunohistochemistry , Lung/chemistry , Lung/drug effects , NF-kappa B/analysis , Ovalbumin , Rats, Wistar , Real-Time Polymerase Chain Reaction , Reproducibility of Results , Treatment Outcome , Vitamins/therapeutic use
5.
Braz. j. microbiol ; 46(1): 285-292, 05/2015. tab, graf
Article in English | LILACS | ID: lil-748256

ABSTRACT

Aspergillus flavus was isolated from soil and exhibited laccase activity under both constitutive and copper induced conditions. Spiking the medium with 1 mM copper sulfate resulted in an increase in the activity which reached 51.84 U/mL, a distinctive protein band was detected at 60 kDa. The extracellular enzyme was purified 81 fold using gel filtration chromatography and resulted in two different laccase fractions L1 and L2, the latter had a higher enzymatic activity which reached 79.57 U/mL and specific activity of 64.17 U/μg protein. The analysis of the spectrum of the L2 fraction showed a shoulder at 330 nm which is characteristic for T2/T3 copper centers; both copper and zinc were detected suggesting that this is an unconventional white laccase. Primers of laccase gene were designed and synthesized to recover specific gene from A. flavus. Sequence analysis indicated putative laccase (Genbank ID: JF683612) at the amino acid level suggesting a close identity to laccases from other genera containing the copper binding site. Decolorization of textile waste water under different conditions showed possible application in bioremediation within a short period of time. The effect of copper on A. flavus was concentration dependent.


Subject(s)
Aspergillus flavus/drug effects , Aspergillus flavus/enzymology , Copper/metabolism , Gene Expression Regulation, Enzymologic/drug effects , Laccase/biosynthesis , Transcriptional Activation/drug effects , Aspergillus flavus/genetics , Aspergillus flavus/isolation & purification , Chromatography, Gel , Culture Media/chemistry , DNA, Fungal/genetics , Electrophoresis, Polyacrylamide Gel , Industrial Waste , Laccase/chemistry , Laccase/isolation & purification , Molecular Sequence Data , Molecular Weight , Sequence Analysis, DNA , Sequence Homology, Amino Acid , Soil Microbiology , Spectrum Analysis , Water Purification
6.
Clinics ; 70(5): 350-355, 05/2015. tab, graf
Article in English | LILACS | ID: lil-748270

ABSTRACT

OBJECTIVE: Sacrococcygeal pilonidal sinus is common in young men and may recur over time after surgery. We investigated whether a factor exists that can aid in the determination of the preferred technique between the early Limberg flap and Karydakis flap techniques for treating recurrent pilonidal sinus. MATERIALS AND METHODS: This prospective and randomized study enrolled 71 patients with recurrent pilonidal sinus in whom the Limberg flap or Karydakis flap techniques were applied for reconstruction after excision. Patients were divided into two groups as follows: 37 patients were treated with the Limberg flap technique and 34 patients were treated with the Karydakis flap technique. Fluid collection, wound infection, flap edema, hematoma, partial wound separation, return to daily activities, pain score, complete healing time, painless seating and patient satisfaction were compared between the groups. ClinicalTrial.gov: NCT02287935. RESULTS: The development rates of total fluid collection, wound infection, flap edema, hematoma, and partial wound separation were 9.8%, 16%, 7%, 15% and 4.2%, respectively; total flap necrosis was not observed in any patient (p<0.001). During the average follow-up of 28 months, no patients (0%) developed recurrent disease. The two groups differed with respect to early surgical complications (p<0.001). CONCLUSION: In this study, use of the Limberg flap was associated with lower complication rates, shorter length of hospital stay, early return to work, low pain score, high patient satisfaction and better complete healing duration. Therefore, we recommend the Limberg flap for treatment of recurrent pilonidal sinus. .


Subject(s)
Animals , Male , Rats , Aryldialkylphosphatase/genetics , Environmental Pollutants/toxicity , Gene Expression Regulation, Enzymologic/drug effects , Liver/drug effects , Polychlorinated Biphenyls/toxicity , Antioxidants/metabolism , Aryldialkylphosphatase/blood , Cholesterol, HDL/blood , Cholesterol, LDL/blood , Dose-Response Relationship, Drug , Lipid Peroxidation/drug effects , Liver/enzymology , Malondialdehyde/metabolism , Rats, Sprague-Dawley , Thiobarbituric Acid Reactive Substances/metabolism
7.
Salud colect ; 11(1): 99-114, ene.-mar. 2015. ilus, tab
Article in Spanish | LILACS | ID: lil-746687

ABSTRACT

El Consejo Federal de Medicina de Brasil (CFM) -órgano normativo y fiscalizador del ejercicio ético de la medicina- prohibió, en 2008, la participación de médicos brasileños en investigaciones que utilizaran placebo para enfermedades con tratamiento eficaz y efectivo, en contraposición a la Declaración de Helsinki, que permite su uso en condiciones metodológicamente justificadas. Con el objetivo de verificar si la normativa ética del CFM modificó el uso de placebo en ensayos clínicos de fase III en Brasil, se analizaron varias características de sus registros en el ClinicalTrials.gov, en los períodos de 2003 a 2007 y de 2009 a 2013. Se concluye que: a) la normativa promulgada por el CFM en 2008 fue ineficaz y prevaleció la posición adoptada por la Declaración de Helsinki; b) el patrocinio de ensayos con placebo por parte de la industria farmacéutica multinacional fue significativo; c) predominaron las investigaciones de fármacos para enfermedades crónicas, y fueron poco significativas para las enfermedades postergadas, de importancia para Brasil.


In 2008, Brazil's Federal Council of Medicine [Conselho Federal de Medicina] (CFM) - regulatory and supervisory agency on the ethical practice of medicine - banned the participation of Brazilian doctors in studies using placebos for diseases with efficient and effective treatment. This position differs with the Helsinki Declaration, which allows the use of placebos in methodologically justified conditions. To ascertain whether the CMF's ethical regulation modified the use of placebos in phase III clinical trials in Brazil, characteristics of the records in ClinicalTrials.gov were researched in the periods from 2003 to 2007 and from 2009 to 2013. The conclusions reached were: a) the regulations issued by the CFM in 2008 were ineffective and the position adopted by the Helsinki Declaration prevails; b) there was significant sponsorship by the multinational pharmaceutical industry of trials with placebos; c) the research was predominantly on new drugs for chronic diseases, with little study done of the neglected diseases which are of great importance to Brazil.


Subject(s)
Animals , Rats , Apoptosis/genetics , Gene Expression Regulation, Enzymologic/genetics , Heme/deficiency , Nerve Degeneration/genetics , Neurons/metabolism , Porphyrias/complications , Apoptosis/drug effects , Caspases/drug effects , Caspases/metabolism , Cell Survival/drug effects , Cell Survival/genetics , Collagen Type XI/drug effects , Collagen Type XI/metabolism , Cyclic AMP Response Element-Binding Protein/drug effects , Cyclic AMP Response Element-Binding Protein/genetics , Cyclic AMP Response Element-Binding Protein/metabolism , Down-Regulation/drug effects , Down-Regulation/physiology , Enzyme Inhibitors , Gene Expression Regulation, Enzymologic/drug effects , Heptanoates , Heme/biosynthesis , MAP Kinase Signaling System/drug effects , MAP Kinase Signaling System/physiology , Membrane Proteins/drug effects , Membrane Proteins/genetics , Membrane Proteins/metabolism , Nerve Degeneration/metabolism , Nerve Degeneration/physiopathology , Nerve Tissue Proteins/drug effects , Nerve Tissue Proteins/genetics , Nerve Tissue Proteins/metabolism , Neural Cell Adhesion Molecules/drug effects , Neural Cell Adhesion Molecules/genetics , Neural Cell Adhesion Molecules/metabolism , Neurons/drug effects , Neurons/pathology , Poly(ADP-ribose) Polymerases , Porphyrias/metabolism , Porphyrias/physiopathology , RNA, Messenger/drug effects , RNA, Messenger/metabolism , RNA-Binding Proteins/drug effects , RNA-Binding Proteins/genetics , RNA-Binding Proteins/metabolism , SMN Complex Proteins , Up-Regulation/drug effects , Up-Regulation/physiology , Vesicular Transport Proteins/drug effects , Vesicular Transport Proteins/genetics , Vesicular Transport Proteins/metabolism
8.
Braz. j. biol ; 74(4): 753-760, 11/2014.
Article in English | LILACS | ID: lil-732297

ABSTRACT

Microcystins (MC) are the most studied toxins of cyanobacteria since they are widely distributed and account for several cases of human and animal poisoning, being potent inhibitors of the serine/threonine protein phosphatases 1 (PP1) and 2A (PP2A). The phosphatases PP1 and PP2A are also present in plants, which may also suffer adverse effects due to the inhibition of these enzymes. In aquatic plants, biomass reduction is usually observed after absorption of cyanotoxins, which can bioaccumulate in its tissues. In terrestrial plants, the effects caused by microcystins vary from inhibition to stimulation as the individuals develop from seedling to adult, and include reduction of protein phosphatases 1 and 2A, oxidative stress, decreased photosynthetic activity and even cell apoptosis, as well as bioaccumulation in plant tissues. Thus, the irrigation of crop plants by water contaminated with microcystins is not only an economic problem but becomes a public health issue because of the possibility of food contamination, and this route of exposure requires careful monitoring by the responsible authorities.


Microcistinas (MC) são as toxinas de cianobactérias mais estudadas, uma vez que são amplamente distribuídas e responsáveis por vários casos de intoxicação humana e animal. São potentes inibidoras das proteínas fosfatases serina/treonina 1 (PP1) e 2A (PP2A). As fosfatases PP1 e PP2A também estão presentes em plantas, as quais podem sofrer efeitos adversos devido à inibição dessas enzimas. Em plantas aquáticas, a redução da biomassa é geralmente observada após absorção de cianotoxinas que podem bioacumular em seus tecidos. Em plantas terrestres, os efeitos causados pelas microcistinas variam de inibição ao estímulo, como no desenvolvimento de plântulas ao estádio adulto, e incluem a redução de proteínas fosfatases 1 e 2A, estresse oxidativo, diminuição da atividade fotossintética e até mesmo apoptose celular, bem como a bioacumulação em tecidos de plantas. Assim, a irrigação de plantas cultivadas com água contaminada com microcistina não é apenas um problema econômico, mas torna-se um problema de saúde pública, devido à possibilidade de contaminação dos alimento, sendo uma via de exposição que requer um monitoramento cuidadoso por parte das autoridades responsáveis.


Subject(s)
Bacterial Toxins/toxicity , Crops, Agricultural/drug effects , Microcystins/toxicity , Crops, Agricultural/enzymology , Environmental Monitoring , Gene Expression Regulation, Enzymologic/drug effects , Protein Phosphatase 1/antagonists & inhibitors , /antagonists & inhibitors
9.
Article in English | WPRIM | ID: wpr-56424

ABSTRACT

To investigate 1alpha,25-(OH)2D3 regulation of matrix metalloproteinase-9 (MMP-9) protein expression during osteoclast formation and differentiation, receptor activator of nuclear factor kappaB ligand (RANKL) and macrophage colony-stimulating factor (M-CSF) were administered to induce the differentiation of RAW264.7 cells into osteoclasts. The cells were incubated with different concentrations of 1alpha,25-(OH)2D3 during culturing, and cell proliferation was measured using the methylthiazol tetrazolium method. Osteoclast formation was confirmed using tartrate-resistant acid phosphatase (TRAP) staining and assessing bone lacunar resorption. MMP-9 protein expression levels were measured with Western blotting. We showed that 1alpha,25-(OH)2D3 inhibited RAW264.7 cell proliferation induced by RANKL and M-CSF, increased the numbers of TRAP-positive osteoclasts and their nuclei, enhanced osteoclast bone resorption, and promoted MMP-9 protein expression in a concentration-dependent manner. These findings indicate that 1alpha,25-(OH)2D3 administered at a physiological relevant concentration promoted osteoclast formation and could regulate osteoclast bone metabolism by increasing MMP-9 protein expression during osteoclast differentiation.


Subject(s)
Acid Phosphatase/metabolism , Animals , Blotting, Western , Calcitriol/pharmacology , Calcium Channel Agonists/pharmacology , Cell Differentiation , Cell Line , Cell Proliferation , Gene Expression Regulation, Enzymologic/drug effects , Isoenzymes/metabolism , Matrix Metalloproteinase 9/genetics , Mice , Osteoclasts/cytology , Tetrazolium Salts , Thiazoles
10.
Article in English | WPRIM | ID: wpr-14962

ABSTRACT

Higher levels of body fat are associated with an increased risk for development numerous adverse health conditions. FTY720 is an immune modulator and a synthetic analogue of sphingosine 1-phosphate (S1P), activated S1P receptors and is effective in experimental models of transplantation and autoimmunity. Whereas immune modulation by FTY720 has been extensively studied, other actions of FTY720 are not well understood. Here we describe a novel role of FTY720 in the prevention of obesity, involving the regulation of adipogenesis and lipolysis in vivo and in vitro. Male C57B/6J mice were fed a standard diet or a high fat diet (HFD) without or with FTY720 (0.04 mg/kg, twice a week) for 6 weeks. The HFD induced an accumulation of large adipocytes, down-regulation of phosphorylated AMP-activated protein kinase alpha (p-AMPKalpha) and Akt (p-Akt); down-regulation of hormone-sensitive lipase (HSL), adipose triglyceride lipase (ATGL) and perilipin mRNA as well as up-regulation of phosphorylated HSL (p-HSL, Ser563) and glycogen synthase kinase 3 alpha/beta (p-GSK3alpha/beta). All these effects were blunted by FTY720 treatment, which inhibited adipogenesis and promoted lipolysis. Also, FTY720 significantly decreased lipid accumulation in maturing preadipocytes. FTY720 down-regulated the transcriptional levels of the PPARgamma, C/EBPalpha and adiponectin, which are markers of adipogenic differentiation. FTY720 significantly increased the release of glycerol and the expression of the HSL, ATGL and perilipin, which are regulators of lipolysis. These results show that FTY720 prevented obesity by modulating adipogenesis and lipolysis, and suggest that FTY720 is used for the treatment of obesity.


Subject(s)
3T3-L1 Cells , AMP-Activated Protein Kinases/metabolism , Adipocytes/drug effects , Adipogenesis/drug effects , Animals , Anti-Obesity Agents/pharmacology , Antigens, Differentiation/genetics , Carrier Proteins/genetics , Cell Size , Diet, High-Fat/adverse effects , Disease Models, Animal , Enzyme Activation , Gene Expression Regulation, Enzymologic/drug effects , Glycogen Synthase Kinase 3/genetics , Lipase/genetics , Lipolysis/drug effects , Male , Mice , Mice, Inbred C57BL , Obesity/etiology , Phosphoproteins/genetics , Phosphorylation , Propylene Glycols/pharmacology , Protein Processing, Post-Translational , Proto-Oncogene Proteins c-akt/metabolism , Sphingosine/analogs & derivatives , Sterol Esterase/metabolism
11.
Article in English | WPRIM | ID: wpr-149765

ABSTRACT

Triptolide, a compound extracted from the traditional Chinese medicine preparation of Tripterygium wilfordii Hook F., has been reported to have anti-inflammatory and anti-cancer activities. However, its effect on ovarian cancer invasion is unknown. We observed that MMP7 and MMP19 expression increased in ovarian cancer tissue. Triptolide treatment inhibited the migration and invasion of ovarian cancer cells SKOV3 and A2780 at the concentration of 15 nM. We also observed that triptolide suppressed MMP7 and MMP19 promoter activity in a dose-dependent manner, down-regulating the expressions of these promoters on mRNA and protein level. Moreover, triptolide enhanced E-cadherin expression in ovarian cancer cells. In vivo, triptolide inhibited tumor formation and metastasis in nude mice, and suppressed MMP7 and MMP19 expression; it also enhanced E-cadherin expression in tumor in a dose-dependent manner. Over expression of MMP7 and MMP19, or suppression of E-cadherin expression partially abolished the inhibitory effect of triptolide on invasion of ovarian cancer cells. To summarize, triptolide significantly inhibited the migration and invasion of ovarian cancer cells by suppression of MMP7 and MMP19 and up-regulation of E-cadherin expression. This study shows that triptolide is a good candidate for the treatment of ovarian cancer and reduction of metastasis.


Subject(s)
Animals , Antineoplastic Agents, Alkylating/pharmacology , Cadherins/genetics , Cell Line, Tumor , Cell Movement/drug effects , Cell Proliferation/drug effects , Cystadenocarcinoma, Serous/drug therapy , Diterpenes/pharmacology , Epoxy Compounds/pharmacology , Female , Gene Expression Regulation, Enzymologic/drug effects , Humans , Matrix Metalloproteinase 7/genetics , Matrix Metalloproteinases, Secreted/genetics , Mice , Mice, Inbred BALB C , Mice, Nude , Neoplasm Invasiveness , Ovarian Neoplasms/drug therapy , Paclitaxel/pharmacology , Phenanthrenes/pharmacology , Promoter Regions, Genetic , Up-Regulation/drug effects , Xenograft Model Antitumor Assays
12.
BCCR-Basic and Clinical Cancer Research. 2011; 3 (1): 14-22
in English | IMEMR | ID: emr-137510

ABSTRACT

The present study was performed to investigate the effects of trace elements particularity Se, Zn and Cu on tumor genesis in breast cancer. The inhibitory effect of Se, Zn and Cu, on telomerase activity was analyzed in human breast tumor tissues and breast cancer [T47D] cells. Tissue specimens from 24 women with benign breast disease and 32 women with breast cancer specimens [ductal carcinoma, lobular carcinoma] were collected during surgery. In addition venous blood samples were obtained for assessing the trace elements. T47D cell line was cultured and treated with trace elements. Telomerase activity then was measured with TRAP assay in cell line and tissue extracts. There was a significant difference between tissue and serum levels of Cu, Se and the ratio of Cu/Zn in patients and controls [P<0.001]. After treating with 100 microm/L Zn So4, 10 um /L Cu So4 for 6 hours, telomerase activity of T47D cells was markedly increased. But after treating with 10, and 30 um /L selenium-L- methionin, telomerase activity was markedly inhibited. Telomerase activity of T47D cells for 24 hours were 0.93, 0.60 and for 48 hours were 0.76, 0.12 respectively [control 49.2%]. There were variations in serum level of Zn and Cu in breast cancer patients. Association between trace elements level and telomerase activity level can be exploited as prognostic and diagnostic marker for breast cancer


Subject(s)
Humans , Female , Breast Neoplasms/enzymology , Telomerase/genetics , Selenium/pharmacology , Zinc/pharmacology , Copper/pharmacology , Cell Line , Tissue Extracts , Tumor Cells, Cultured , Gene Expression Regulation, Enzymologic/drug effects , Gene Expression Regulation, Neoplastic/drug effects
13.
Yonsei Medical Journal ; : 653-660, 2010.
Article in English | WPRIM | ID: wpr-46865

ABSTRACT

PURPOSE: The ketogenic diet has long been used to treat epilepsy, but its mechanism is not yet clearly understood. To explore the potential mechanism, we analyzed the changes in gene expression induced by the ketogenic diet in the rat kainic acid (KA) epilepsy model. MATERIALS AND METHODS: KA-administered rats were fed the ketogenic diet or a normal diet for 4 weeks, and microarray analysis was performed with their brain tissues. The effects of the ketogenic diet on cathepsin E messenger ribonucleic acid (mRNA) expression were analyzed in KA-administered and normal saline-administered groups with semi-quantitative and real-time reverse transcription polymerase chain reaction (RT-PCR). Brain tissues were dissected into 8 regions to compare differential effects of the ketogenic diet on cathepsin E mRNA expression. Immunohistochemistry with an anti-cathepsin E antibody was performed on slides of hippocampus obtained from whole brain paraffin blocks. RESULTS: The microarray data and subsequent RT-PCR experiments showed that KA increased the mRNA expression of cathepsin E, known to be related to neuronal cell death, in most brain areas except the brain stem, and these increases of cathepsin E mRNA expression were suppressed by the ketogenic diet. The expression of cathepsin E mRNA in the control group, however, was not significantly affected by the ketogenic diet. The change in cathepsin E mRNA expression was greatest in the hippocampus. The protein level of cathepsin E in the hippocampus of KA-administered rat was elevated in immunohistochemistry and the ketogenic diet suppressed this increase. CONCLUSION: Our results showed that KA administration increased cathepsin E expression in the rat brain and its increase was suppressed by the ketogenic diet.


Subject(s)
3-Hydroxybutyric Acid/blood , Animals , Cathepsin E/genetics , Enzyme Activators/pharmacology , Gene Expression Regulation, Enzymologic/drug effects , Hippocampus/drug effects , Immunohistochemistry , Kainic Acid/pharmacology , Diet, Ketogenic , Male , Oligonucleotide Array Sequence Analysis , Rats , Rats, Sprague-Dawley , Reverse Transcriptase Polymerase Chain Reaction
14.
Article in English | WPRIM | ID: wpr-27757

ABSTRACT

As glucose is known to induce insulin secretion in pancreatic beta cells, this study investigated the role of a phospholipase D (PLD)-related signaling pathway in insulin secretion caused by high glucose in the pancreatic beta-cell line MIN6N8. It was found that the PLD activity and PLD1 expression were both increased by high glucose (33.3 mM) treatment. The dominant negative PLD1 inhibited glucose-induced Beta2 expression, and glucose-induced insulin secretion was blocked by treatment with 1-butanol or PLD1-siRNA. These results suggest that high glucose increased insulin secretion through a PLD1-related pathway. High glucose induced the binding of Arf6 to PLD1. Pretreatment with brefeldin A (BFA), an Arf inhibitor, decreased the PLD activity as well as the insulin secretion. Furthermore, BFA blocked the glucose-induced mTOR and p70S6K activation, while mTOR inhibition with rapamycin attenuated the glucose induced Beta2 expression and insulin secretion. Thus, when taken together, PLD1 would appear to be an important regulator of glucose-induced insulin secretion through an Arf6/PLD1/mTOR/p70S6K/Beta2 pathway in MIN6N8 cells.


Subject(s)
ADP-Ribosylation Factors/metabolism , Animals , Basic Helix-Loop-Helix Transcription Factors/metabolism , Cells, Cultured , Gene Expression Regulation, Enzymologic/drug effects , Glucose/pharmacology , Insulin/metabolism , Insulin-Secreting Cells/drug effects , Intracellular Signaling Peptides and Proteins/metabolism , Mice , Models, Biological , Oligodeoxyribonucleotides, Antisense/pharmacology , Phospholipase D/antagonists & inhibitors , Protein-Serine-Threonine Kinases/metabolism , Ribosomal Protein S6 Kinases, 70-kDa/metabolism , Signal Transduction/drug effects
15.
Article in English | WPRIM | ID: wpr-203590

ABSTRACT

The action mode of 4,4'-diaminodiphenylsulfone (DDS) is still under debate, although it has long been used in treatment of several dermatologic diseases including Hansen's disease. In this study, we tested the effect of DDS as an antioxidant on paraquat-induced oxidative stress in non-phagocytic human diploid fibroblasts (HDFs). Overall, preincubation of HDFs with DDS prevented the oxidative stress and the resulting cytotoxic damages caused by paraquat in these cells. The specific effects of DDS in paraquat-treated HDFs are summarized as follows: a) reducing the expression of NADPH oxidase 4 (NOX4) by inhibiting paraquat-induced activation of PKC; b) inhibiting paraquat-induced decreases in mitochondrial complex protein levels as well as in membrane potentials; c) consequently, inhibiting the generation of cytosolic and mitochondrial superoxide anions. Taken together, these findings suggest that DDS would suppress the radical generation in non-phagocytic HDFs during oxidative stress, and that DDS might have the extended potential to be used further in prevention of other oxidative stress-related pathologies.


Subject(s)
Biphenyl Compounds/metabolism , Cell Death/drug effects , Cell Survival/drug effects , Dapsone/pharmacology , Diploidy , Enzyme Activation/drug effects , Fibroblasts/cytology , Free Radical Scavengers/metabolism , Gene Expression Regulation, Enzymologic/drug effects , Humans , Male , Mitochondria/drug effects , NADPH Oxidases/genetics , Paraquat/toxicity , Phagocytosis/drug effects , Picrates/metabolism , Protein Kinase C/metabolism , RNA, Messenger/genetics , Reactive Oxygen Species/metabolism , Superoxides/metabolism
16.
Article in English | WPRIM | ID: wpr-49339

ABSTRACT

Matrix metalloproteinase-9 (MMP-9) may play an important role in emphysematous change in chronic obstructive pulmonary disease (COPD), one of the leading causes of mortality and morbidity worldwide. We previously reported that simvastatin, an inhibitor of HMG-CoA reductase, attenuates emphysematous change and MMP-9 induction in the lungs of rats exposed to cigarette smoke. However, it remained uncertain how cigarette smoke induced MMP-9 and how simvastatin inhibited cigarette smoke-induced MMP-9 expression in alveolar macrophages (AMs), a major source of MMP-9 in the lungs of COPD patients. Presently, we examined the related signaling for MMP-9 induction and the inhibitory mechanism of simvastatin on MMP-9 induction in AMs exposed to cigarette smoke extract (CSE). In isolated rat AMs, CSE induced MMP-9 expression and phosphorylation of ERK and Akt. A chemical inhibitor of MEK1/2 or PI3K reduced phosphorylation of ERK or Akt, respectively, and also inhibited CSE-mediated MMP-9 induction. Simvastatin reduced CSE-mediated MMP-9 induction, and simvastatin-mediated inhibition was reversed by farnesyl pyrophosphate (FPP) or geranylgeranyl pyrophosphate (GGPP). Similar to simvastatin, inhibition of FPP transferase or GGPP transferase suppressed CSE-mediated MMP-9 induction. Simvastatin attenuated CSE-mediated activation of RAS and phosphorylation of ERK, Akt, p65, IkappaB, and nuclear AP-1 or NF-kappaB activity. Taken together, these results suggest that simvastatin may inhibit CSE-mediated MMP-9 induction, primarily by blocking prenylation of RAS in the signaling pathways, in which Raf-MEK-ERK, PI3K/Akt, AP-1, and IkappaB-NF-kappaB are involved.


Subject(s)
Phosphatidylinositol 3-Kinase/metabolism , Alkyl and Aryl Transferases/metabolism , Animals , Anticholesteremic Agents/pharmacology , Cells, Cultured , Enzyme Inhibitors/metabolism , Extracellular Signal-Regulated MAP Kinases/metabolism , Gene Expression Regulation, Enzymologic/drug effects , I-kappa B Kinase/antagonists & inhibitors , Macrophages, Alveolar/cytology , Matrix Metalloproteinase 9/genetics , Mitogen-Activated Protein Kinase Kinases/metabolism , Polyisoprenyl Phosphates/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Rats , Sesquiterpenes/metabolism , Signal Transduction/physiology , Simvastatin/pharmacology , Smoke/adverse effects , Tobacco/adverse effects
17.
Indian J Physiol Pharmacol ; 2008 Jan-Mar; 52(1): 11-8
Article in English | IMSEAR | ID: sea-107104

ABSTRACT

Nitric oxide (NO) is formed by different cell types in response to a variety of physiological and patho-physiological stimuli. The intake of nicotine and/or alcohol has patho-physiological effects on organ function, and the progression of alcohol-/tobacco-related diseases seem to be directly influenced by NO-mediated mechanisms. Nicotine has an adverse influence on blood vessel functionality, repair and maintenance. Chronic nicotine exposure augments atherosclerosis by enhancing the production of proinflammatory cytokines by macrophages which then activate atherogenic NF-kB target genes in aortic lesions. Alcohol produces NO which speeds up the apoptosis of neutrophils. Alcohol sensitizes the liver to endotoxemic shock. Nitrosative stress and increased basal levels of NO contribute to tumour growth. The progression of disease seems to be directed via a definite NO-mediated mechanism. This review gives an insight into how intake of tobacco and alcohol may affect quality of life.


Subject(s)
Animals , Central Nervous System Depressants/toxicity , Drug Interactions , Ethanol/toxicity , Gene Expression Regulation, Enzymologic/drug effects , Humans , Neoplasms/etiology , Nicotine/toxicity , Nicotinic Agonists/toxicity , Nitric Oxide/physiology , Nitric Oxide Synthase/genetics , Oxidative Stress/drug effects
18.
Article in English | WPRIM | ID: wpr-57370

ABSTRACT

Quercetin 3-O-beta-(2''-galloyl)-rhamnopyranoside (QGR) is a naturally occurring quercitrin gallate, which is a polyphenolic compound that was originally isolated from Persicaria lapathifolia (Polygonaceae). QGR has been shown to have an inhibitory effect on nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated macrophage RAW 264.7 cells. Therefore, this study was conducted to investigate the inhibitory effect of QGR on nitric oxide production and inducible nitric oxide synthases (iNOS) expression in LPS-stimulated Balb/c mice. To accomplish this, 10 mg/kg of QGR was administered via gavage once a day for 3 days. iNOS was then induced by intraperitoneal injection of LPS. Six hours after the LPS treatment the animals were sacrificed under ether anethesia. The serum levels of NO were then measured to determine if QGR exerted an inhibitory effect on NO production in vivo. LPS induced an approximately 6 fold increase in the expression of NO. However, oral administration of QGR reduced the LPS induced increase in NO by half. Furthermore, RT-PCR and western blot analysis revealed that the increased levels of iNOS expression that occurred in response to treatment with LPS were significantly attenuated in response to QGR pretreatment. Histologically, LPS induced the infiltration of polymorphonuclear neutrophils in portal veins and sinusoids and caused the formation of a large number of necrotic cells; however, pretreatment with QGR attenuated these LPS induced effects. Taken together, these results indicate that QGR inhibits iNOS expression in vivo as well as in vitro and has antiinflammatory potentials.


Subject(s)
Animals , DNA Primers , Gene Expression Regulation, Enzymologic/drug effects , Lipopolysaccharides/pharmacology , Liver/drug effects , Mice , Mice, Inbred BALB C , Nitric Oxide/blood , Nitric Oxide Synthase Type II/drug effects , Quercetin/analogs & derivatives , RNA, Messenger/genetics , Reverse Transcriptase Polymerase Chain Reaction
19.
Article in English | WPRIM | ID: wpr-171132

ABSTRACT

We investigated the mechanism of spontaneous cholesterol efflux induced by acyl-coenzyme A:cholesterol acyltransferase (ACAT) inhibition, and how an alteration of cholesterol metabolism in macrophages impacts on that in HepG2 cells. Oleic acid anilide (OAA), a known ACAT inhibitor reduced lipid storage substantially by promotion of cholesterol catabolism and repression of cholesteryl ester accumulation without further increase of cytotoxicity in acetylated low-density lipoprotein-loaded THP-1 macrophages. Analysis of expressed mRNA and protein revealed that cholesterol 7alpha-hydroxylase (CYP7A1), oxysterol 7alpha- hydroxylase (CYP7B1), and cholesterol 27-hydroxylase (CYP27) were highly induced by ACAT inhibition. The presence of a functional cytochrome P450 pathway was confirmed by quantification of the biliary cholesterol mass in cell monolayers and extracelluar medium. Notably, massively secreted biliary cholesterol from macrophages suppressed the expression of CYP7 proteins in a farnesoid X receptor (FXR)-dependent manner in HepG2 cells. The findings reported here provide new insight into mechanisms of spontaneous cholesterol efflux, and suggest that ACAT inhibition may stimulate cholesterol-catabolic (cytochrome P450) pathway in lesion-macrophages, in contrast, suppress it in hepatocyte via FXR induced by biliary cholesterol (BC).


Subject(s)
Anilides/pharmacology , Bile/metabolism , Cells, Cultured , Cholesterol/metabolism , Cholesterol Esters/metabolism , DNA-Binding Proteins/agonists , Enzyme Inhibitors/pharmacology , Gene Expression Regulation, Enzymologic/drug effects , Hepatocytes/drug effects , Humans , Lipid Metabolism/drug effects , Macrophages/drug effects , Models, Biological , Oleic Acids/pharmacology , Receptors, Cytoplasmic and Nuclear/agonists , Sterol O-Acyltransferase/antagonists & inhibitors , Transcription Factors/agonists
20.
Article in English | WPRIM | ID: wpr-90619

ABSTRACT

Curcumin is a polyphenolic compound possessing interesting anti-inflammatory and antioxidant properties and has the ability to induce the defensive protein heme oxygenase-1 (HO-1). The objective of this study was to investigate whether curcumin protects against cold storage-mediated damage of human adult atrial myoblast cells (Girardi cells) and to assess the potential involvement of HO-1 in this process. Girardi cells were exposed to either normothermic or hypothermic conditions in Celsior preservation solution in the presence or absence of curcumin. HO-1 protein expression and heme oxygenase activity as well as cellular damage were assessed after cold storage or cold storage followed by re-warming. In additional experiments, an inhibitor of heme oxygenase activity (tin protoporphyrin IX, micrometer) or siRNA for HO-1 were used to investigate the participation of HO-1 as a mediator of curcumin- induced effects. Treatment with curcumin produced a marked induction of cardiac HO-1 in normothermic condition but cells were less responsive to the polyphenolic compound at low temperature. Cold storage-induced damage was markedly reduced in the presence of curcumin and HO-1 contributed to some extent to this effect. Thus, curcumin added to Celsior preservation solution effectively prevents the damage caused by cold- storage; this effect involves the protective enzyme HO-1 but also other not yet identified mechanisms.


Subject(s)
Cell Death/drug effects , Cell Survival/drug effects , Cells, Cultured , Cold Temperature , Cryopreservation , Cryoprotective Agents/pharmacology , Curcumin/pharmacology , Gene Expression Regulation, Enzymologic/drug effects , Heme Oxygenase-1/genetics , Hemin/pharmacology , Humans , Hydrogen Peroxide/pharmacology , Myoblasts, Cardiac/drug effects , RNA, Messenger/genetics
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