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1.
World J Microbiol Biotechnol ; 35(8): 130, 2019 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-31385043

RESUMO

Bacterial biofilms (BFs) are membrane-like structures formed by the secretion of extracellular polymeric substances (EPS) by bacteria. The formation of BFs contributes to bacterial survival and drug resistance. When bacteria proliferate, they produce secondary metabolites that act as signaling molecules in bacterial communities that regulate intracellular and cell-to-cell communication. This communication can directly affect the physiological behavior of bacteria, including the production and emission of light (bioluminescence), the expression of virulence factors, the resistance to antibiotics, and the shift between planktonic and biofilm lifestyles. We review the major signaling molecules that regulate BF formation, with a focus on quorum-sensing systems (QS), cyclic diguanylate (c-di-GMP), two-component systems (TCS), and small RNA (sRNA). Understanding these processes will lead to new approaches for treating chronic diseases and preventing bacterial resistance.


Assuntos
Bactérias/crescimento & desenvolvimento , Biofilmes/crescimento & desenvolvimento , Regulação Bacteriana da Expressão Gênica , Transdução de Sinais , Bactérias/metabolismo , GMP Cíclico/análogos & derivados , GMP Cíclico/metabolismo , Percepção de Quorum , RNA Bacteriano/metabolismo , Pequeno RNA não Traduzido/metabolismo
2.
Chem Biol Interact ; 309: 108710, 2019 Aug 25.
Artigo em Inglês | MEDLINE | ID: mdl-31199930

RESUMO

Formic acid is a common organic acid used in many industrial processes. There is a paucity of research on the direct toxicity of formic acid and how it might affect the cardiovascular system. This study aimed to understand the effect of formic acid on vascular tension in an animal model and the underlying mechanism. Results found that the vasodilation induced by formic acid was related to the endothelium. When the dosage of formic acid was 1 mM or 5 mM, the vasodilation of endothelium-intact rings was partially suppressed by l-NAME, NS-2028 and nifedipine, and vasoconstriction caused by CaCl2 was inhibited, and the mRNA levels of eNOS, the activity of NOS (tNOS, iNOS and cNOS) and the level of NO and cGMP were significantly increased. Results also found that eNOS protein expression was significantly enhanced by 5 mM of formic acid. These results suggest formic acid can relax the aortic vessels of rats in a dose-dependent pattern. Further, the mechanism of the formic acid-induced vasodilatation likely involved the NO/cGMP pathway. Finally, the current study has revealed that vasodilation induced by high concentrations of formaldehyde may be the effect of the metabolite formic acid. This study will help further inform the etiologies of formic acid-related angiocardiopathies.


Assuntos
Endotélio Vascular/efeitos dos fármacos , Formiatos/farmacologia , Transdução de Sinais/efeitos dos fármacos , Regulação para Cima/efeitos dos fármacos , Animais , Canais de Cálcio Tipo L/metabolismo , Cloreto de Cálcio/farmacologia , GMP Cíclico/metabolismo , Endotélio Vascular/fisiologia , Expressão Gênica/efeitos dos fármacos , Masculino , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase/genética , Óxido Nítrico Sintase/metabolismo , Ratos , Ratos Wistar , Vasoconstrição/efeitos dos fármacos , Vasodilatação/efeitos dos fármacos
3.
Cell Mol Biol (Noisy-le-grand) ; 65(4): 83-89, 2019 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-31078160

RESUMO

The follicle must fulfill the following criteria if it is to survive the period between early embryonic life and the luteinizing hormone (LH) peak. It should (i) be surrounded by pregranulosa cells; (ii) complete the first meiotic division and become dormant; and (iii) continue metabolism during the dormant stage. Interaction between the natriuretic peptide precursor type C (Nppc) and its receptor, natriuretic peptide receptor 2 (Npr2), affects female fertility through the production of oocytes with developmental capacity and maintain oocyte meiotic arrest. While Nppc is expressed in mural cells, cumulus cells express Npr2. Nppc/Npr2 system exerts its biological function on developing follicles by increasing the production of intracellular cyclic guanosine monophosphate (cGMP). This pathway not only contributes to the development of ovary and the uterus, but aids the formation of healthy eggs in terms of their morphological and genetic aspects. A defect in this pathway leads to asmall ovarian size, string-like uterine horns, and thin endometrium and myometrium. Disorganized chromosomes, abnormal cumulus expansion and early meiotic resumption occur in animals with defective Nppc/Npr2 signaling. The types and number of oocytes also decrease when there is incompetent Nppc/Npr2 signaling. This paper extends on most recent and relevant experimental evidence regarding Nppc/Npr2/cGMP signaling with regard to its crucial role in maintaining oocyte meiotic arrest and the production of oocytes with developmental capacity. We further discuss whether the agonist or antagonist forms of the members of this exciting pathway can be usedfor triggering final oocyte maturation.


Assuntos
GMP Cíclico/metabolismo , Peptídeo Natriurético Tipo C/metabolismo , Oócitos/metabolismo , Receptores do Fator Natriurético Atrial/metabolismo , Transdução de Sinais , Animais , Fertilização In Vitro , Humanos
4.
PLoS Pathog ; 15(5): e1007670, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-31121005

RESUMO

To elicit effective invasion and egress from infected cells, obligate intracellular parasites of the phylum Apicomplexa rely on the timely and spatially controlled exocytosis of specialized secretory organelles termed the micronemes. The effector molecules and signaling events underpinning this process are intricate; however, recent advances within the field of Toxoplasma gondii research have facilitated a broader understanding as well as a more integrated view of this complex cascade of events and have unraveled the importance of phosphatidic acid (PA) as a lipid mediator at multiple steps in this process.


Assuntos
Cálcio/metabolismo , GMP Cíclico/metabolismo , Exocitose/fisiologia , Organelas/metabolismo , Ácidos Fosfatídicos/metabolismo , Toxoplasma/fisiologia , Toxoplasmose/parasitologia , Animais , Interações Hospedeiro-Parasita , Humanos , Organelas/parasitologia , Transporte Proteico , Proteínas de Protozoários/metabolismo , Transdução de Sinais
5.
Life Sci ; 230: 150-161, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31125564

RESUMO

Despite novel technologies, colon cancer remains undiagnosed and 25% of patients are diagnosed with metastatic colon cancer. Resistant to chemotherapeutic agents is one of the major problems associated with treating colon cancer which creates the need to develop novel agents targeting towards newer targets. A phosphodiesterase is a group of isoenzyme, which, hydrolyze cyclic nucleotides and thereby lowers intracellular levels of cAMP and cGMP leading to tumorigenic effects. Many in vitro and in vivo studies have confirmed increased PDE expression in different types of cancers including colon cancer. cAMP-specific PDE inhibitors increase intracellular cAMP that leads to activation of effector molecules-cAMP-dependent protein kinase A, exchange protein activated by cAMP and cAMP gated ion channels. These molecules regulate cellular responses and exert its anticancer role through different mechanisms including apoptosis, inhibition of angiogenesis, upregulating tumor suppressor genes and suppressing oncogenes. On the other hand, cGMP specific PDE inhibitors exhibit anticancer effects through cGMP dependent protein kinase and cGMP dependent cation channels. Elevation in cGMP works through activation of caspases, suppression of Wnt/b-catenin pathway and TCF transcription leading to inhibition of CDK and survivin. These studies point out towards the fact that PDE inhibition is associated with anti-proliferative, anti-apoptotic and anti-angiogenic pathways involved in its anticancer effects in colon cancer. Thus, inhibition of PDE enzymes can be used as a novel approach to treat colon cancer. This review will focus on cAMP and cGMP signaling pathways leading to tumorigenesis and the use of PDE inhibitors in colon cancer.


Assuntos
Neoplasias do Colo/terapia , Inibidores de Fosfodiesterase/farmacologia , Inibidores de Fosfodiesterase/uso terapêutico , Animais , Apoptose/efeitos dos fármacos , AMP Cíclico/metabolismo , GMP Cíclico/metabolismo , Humanos , Diester Fosfórico Hidrolases/metabolismo , Transdução de Sinais/efeitos dos fármacos
6.
Toxicol Lett ; 312: 55-64, 2019 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-30974163

RESUMO

Formaldehyde (FA), a well-known toxic gas molecule similar to nitric oxide (NO), carbon monoxide (CO), and hydrogen sulfide (H2S), is widely produced endogenously via numerous biochemical pathways, and has a number of physiological roles in the biosystem. We attempted to investigate the vasorelaxant effects of FA and their underlying mechanisms. We found that FA induced vasorelaxant effects on rat aortic rings in a concentration-dependent manner. The NO/cyclic guanosine 5' monophosphate (cGMP) pathway was up-regulated when the rat aortas were treated with FA. The expression of large-conductance Ca2+-activated K+ (BKCa) channel subunits α and ß of the rat aortas was increased by FA. Similarly, the levels of ATP-sensitive K+ (KATP) channel subunits Kir6.1 and Kir6.2 were also up-regulated when the rat aortas were incubated with FA. In contrast, levels of the L-type Ca2+ channel (LTCC) subunits, Cav1.2 and Cav1.3, decreased dramatically with increasing concentrations of FA. We demonstrated that the regulation of FA on vascular contractility may be via the up-regulation of the NO/cGMP pathway and the modulation of ion channels, including the upregulated expression of the KATP and BKCa channels and the inhibited expression of LTCCs. Further study is needed to explore the in-depth mechanisms of FA induced vasorelaxation.


Assuntos
GMP Cíclico/metabolismo , Formaldeído/farmacologia , Óxido Nítrico/metabolismo , Transdução de Sinais/efeitos dos fármacos , Vasodilatação/efeitos dos fármacos , Animais , Aorta Torácica , Canais de Cálcio Tipo L/genética , Canais de Cálcio Tipo L/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Canais KATP/genética , Canais KATP/metabolismo , Subunidades alfa do Canal de Potássio Ativado por Cálcio de Condutância Alta/genética , Subunidades alfa do Canal de Potássio Ativado por Cálcio de Condutância Alta/metabolismo , Masculino , Ratos , Ratos Wistar
7.
MBio ; 10(2)2019 04 23.
Artigo em Inglês | MEDLINE | ID: mdl-31015332

RESUMO

Vibrio cholerae biofilm formation and associated motility suppression are correlated with increased concentrations of cyclic diguanylate monophosphate (c-di-GMP), which are in turn driven by increased levels and/or activity of diguanylate cyclases (DGCs). To further our understanding of how c-di-GMP modulators in V. cholerae individually and collectively influence motility with cellular resolution, we determined how DGCs CdgD and CdgH impact intracellular c-di-GMP levels, motility, and biofilm formation. Our results indicated that CdgH strongly influences swim speed distributions; cells in which cdgH was deleted had higher average swim speeds than wild-type cells. Furthermore, our results suggest that CdgD, rather than CdgH, is the dominant DGC responsible for postattachment c-di-GMP production in biofilms. Lipopolysaccharide (LPS) biosynthesis genes were found to be extragenic bypass suppressors of the motility phenotypes of strains ΔcdgD and ΔcdgH We compared the motility regulation mechanism of the DGCs with that of Gmd, an LPS O-antigen biosynthesis protein, and discovered that comodulation of c-di-GMP levels by these motility effectors can be positively or negatively cooperative rather than simply additive. Taken together, these results suggest that different environmental and metabolic inputs orchestrate DGC responses of V. cholerae via c-di-GMP production and motility modulation.IMPORTANCE Cyclic diguanylate monophosphate (c-di-GMP) is a broadly conserved bacterial signaling molecule that affects motility, biofilm formation, and virulence. Although it has been known that high intracellular concentrations of c-di-GMP correlate with motility suppression and biofilm formation, how the 53 predicted c-di-GMP modulators in Vibrio cholerae collectively influence motility is not understood in detail. Here we used a combination of plate assays and single-cell tracking methods to correlate motility and biofilm formation outcomes with specific enzymes involved in c-di-GMP synthesis in Vibrio cholerae, the causative agent of the disease cholera.


Assuntos
Biofilmes/crescimento & desenvolvimento , GMP Cíclico/análogos & derivados , Proteínas de Escherichia coli/metabolismo , Fósforo-Oxigênio Liases/metabolismo , Vibrio cholerae/enzimologia , Vibrio cholerae/fisiologia , GMP Cíclico/metabolismo , Proteínas de Escherichia coli/genética , Deleção de Genes , Regulação Bacteriana da Expressão Gênica , Locomoção , Fósforo-Oxigênio Liases/genética , Vibrio cholerae/genética
8.
BMC Complement Altern Med ; 19(1): 79, 2019 Apr 02.
Artigo em Inglês | MEDLINE | ID: mdl-30940120

RESUMO

BACKGROUND: Methanol extract (MECN) of Clinacanthus nutans Lindau leaves (family Acanthaceae) demonstrated peripherally and centrally mediated antinociceptive activity via the modulation of opioid/NO-mediated, but cGMP-independent pathway. In the present study, MECN was sequentially partitioned to obtain petroleum ether extract of C. nutans (PECN), which was subjected to antinociceptive study with aims of establishing its antinociceptive potential and determining the role of opioid receptors and L-arginine/nitric oxide/cyclic-guanosine monophosphate (L-arg/NO/cGMP) pathway in the observed antinociceptive activity. METHODS: The antinociceptive potential of orally administered PECN (100, 250, 500 mg/kg) was studied using the abdominal constriction-, hot plate- and formalin-induced paw licking-test in mice (n = 6). The effect of PECN on locomotor activity was also evaluated using the rota rod assay. The role of opioid receptors was determined by pre-challenging 500 mg/kg PECN (p.o.) with antagonist of opioid receptor subtypes, namely ß-funaltrexamine (ß-FNA; 10 mg/kg; a µ-opioid antagonist), naltrindole (NALT; 1 mg/kg; a δ-opioid antagonist) or nor-binaltorphimine (nor-BNI; 1 mg/kg; a κ-opioid antagonist) followed by subjection to the abdominal constriction test. In addition, the role of L-arg/NO/cGMP pathway was determined by prechallenging 500 mg/kg PECN (p.o.) with L-arg (20 mg/kg; a NO precursor), 1H-[1, 2, 4] oxadiazolo [4,3-a]quinoxalin-1-one (ODQ; 2 mg/kg; a specific soluble guanylyl cyclase inhibitor), or the combinations thereof (L-arg + ODQ) for 5 mins before subjection to the abdominal constriction test. PECN was also subjected to phytoconstituents analyses. RESULTS: PECN significantly (p < 0.05) inhibited nociceptive effect in all models in a dose-dependent manner. The highest dose of PECN (500 mg/kg) also did not significantly (p > 0.05) affect the locomotor activity of treated mice. The antinociceptive activity of PECN was significantly (p < 0.05) inhibited by all antagonists of µ-, δ-, and κ-opioid receptors. In addition, the antinociceptive activity of PECN was significantly (p < 0.05) reversed by L-arg, but insignificantly (p > 0.05) affected by ODQ. HPLC analysis revealed the presence of at least cinnamic acid in PECN. CONCLUSION: PECN exerted antinocicpetive activity at peripheral and central levels possibly via the activation of non-selective opioid receptors and modulation of the NO-mediated/cGMP-independent pathway partly via the synergistic action of phenolic compounds.


Assuntos
Acanthaceae/química , Analgésicos/farmacologia , GMP Cíclico/metabolismo , Óxido Nítrico/metabolismo , Extratos Vegetais/farmacologia , Receptores Opioides/efeitos dos fármacos , Alcanos , Analgésicos/química , Animais , Masculino , Camundongos , Camundongos Endogâmicos ICR , Extratos Vegetais/química , Folhas de Planta/química , Receptores Opioides/metabolismo , Transdução de Sinais/efeitos dos fármacos
9.
Cell Physiol Biochem ; 52(4): 922-934, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30964609

RESUMO

BACKGROUND/AIMS: In a previous study, we reported that cardiomyocytes were equipped with non-neuronal cardiac cholinergic system (NNCCS) to synthesize acetylcholine (ACh), which is indispensable for maintaining the basic physiological cardiac functions. The aim of this study was to identify and characterize a pharmacological inducer of NNCCS. METHODS: To identify a pharmacological inducer of NNCCS, we screened several chemical compounds with chemical structures similar to the structure of S-nitroso-N-acetyl-DL-penicillamine (SNAP). Preliminary investigation revealed that SNAP is an inducer of non-neuronal ACh synthesis. We screened potential pharmacological inducers in H9c2 and HEK293 cells using western blot analysis, luciferase assay, and measurements of intracellular cGMP, NO2 and ACh levels. The effects of the screened compound on cardiac function of male C57BL6 mice were also evaluated using cardiac catheter system. RESULTS: Among the tested compounds, we selected S-nitroso-Npivaloyl-D-penicillamine (SNPiP), which gradually elevated the intracellular cGMP levels and nitric oxide (NO) levels in H9c2 and HEK293 cells. These elevated levels resulted in the gradual transactivation and translation of the choline acetyltransferase gene. Additionally, in vitro and in vivo SNPiP treatment elevated ACh levels for 72 h. SNPiP-treated mice upregulated their cardiac function without tachycardia but with enhanced diastolic function resulting in improved cardiac output. The effect of SNPiP was dependent on SNPiP nitroso group as verified by the ineffectiveness of N-pivaloyl-D-penicillamine (PiP), which lacks the nitroso group. CONCLUSION: SNPiP is identified to be one of the important pharmacological candidates for induction of NNCCS.


Assuntos
Acetilcolina/biossíntese , Débito Cardíaco/efeitos dos fármacos , GMP Cíclico/metabolismo , Miócitos Cardíacos/metabolismo , Doadores de Óxido Nítrico , Sistema Colinérgico não Neuronal/efeitos dos fármacos , Animais , Células HEK293 , Humanos , Masculino , Camundongos , Óxido Nítrico/biossíntese , Doadores de Óxido Nítrico/química , Doadores de Óxido Nítrico/farmacologia
10.
Molecules ; 24(7)2019 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-30965668

RESUMO

To test whether KMUP-1 (7-[2-[4-(2-chlorophenyl) piperazinyl]ethyl]-1,3-dimethylxanthine) prevents myocardial ischemia-induced apoptosis, we examined KMUP-1-treated H9c2 cells culture. Recent attention has focused on the activation of nitric oxide (NO)-guanosine 3', 5'cyclic monophosphate (cGMP)-protein kinase G (PKG) signaling pathway triggered by mitogen-activated protein kinase (MAPK) family, including extracellular-signal regulated kinase 1/2 (ERK1/2), c-Jun N-terminal kinase (JNK), and p38 in the mechanism of cardiac protection during ischemia-induced cell-death. We propose that KMUP-1 inhibits ischemia-induced apoptosis in H9c2 cells culture through these pathways. Cell viability was assessed using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and apoptotic evaluation was conducted using DNA ladder assay and Hoechst 33342 staining. The level of intracellular calcium was detected using - Fura2-acetoxymethyl (Fura2-AM) staining, and mitochondrial calcium with Rhod 2-acetoxymethyl (Rhod 2-AM) staining under fluorescence microscopic observation. The expression of endothelium NO synthase (eNOS), inducible NO synthase (iNOS), soluble guanylate cyclase α1 (sGCα1), PKG, Bcl-2/Bax ratio, ERK1/2, p38, and JNK proteins were measured by Western blotting assay. KMUP-1 pretreatment improved cell viability and inhibited ischemia-induced apoptosis of H9c2 cells. Calcium overload both in the intracellular and mitochondrial sites was attenuated by KMUP-1 pretreatment. Moreover, KMUP-1 reduced intracellular reactive oxygen species (ROS), increased plasma NOx (nitrite and nitrate) level, and the expression of eNOS. Otherwise, the iNOS expression was downregulated. KMUP-1 pretreatment upregulated the expression of sGCα1 and PKG protein. The ratio of Bcl-2/Bax expression was increased by the elevated level of Bcl2 and decreased level of Bax. In comparison with the ischemia group, KMUP-1 pretreatment groups reduced the expression of phosphorylated extracellular signal-regulated kinases ERK1/2, p-p38, and p-JNK as well. Therefore, KMUP-1 inhibits myocardial ischemia-induced apoptosis by restoration of cellular calcium influx through the mechanism of NO-cGMP-MAPK pathways.


Assuntos
Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Isquemia Miocárdica/metabolismo , Miócitos Cardíacos/citologia , Piperidinas/farmacologia , Xantinas/farmacologia , Animais , Apoptose/efeitos dos fármacos , Cálcio/metabolismo , Linhagem Celular , GMP Cíclico/metabolismo , Mitocôndrias/metabolismo , Modelos Biológicos , Isquemia Miocárdica/tratamento farmacológico , Miócitos Cardíacos/efeitos dos fármacos , Miócitos Cardíacos/metabolismo , Óxido Nítrico/metabolismo , Ratos
11.
Proc Natl Acad Sci U S A ; 116(13): 6335-6340, 2019 03 26.
Artigo em Inglês | MEDLINE | ID: mdl-30862737

RESUMO

Salmonella Typhimurium can invade and survive within macrophages where the bacterium encounters a range of host environmental conditions. Like many bacteria, S. Typhimurium rapidly responds to changing environments by the use of second messengers such as cyclic di-GMP (c-di-GMP). Here, we generate a fluorescent biosensor to measure c-di-GMP concentrations in thousands of individual bacteria during macrophage infection and to define the sensor enzymes important to c-di-GMP regulation. Three sensor phosphodiesterases were identified as critical to maintaining low c-di-GMP concentrations generated after initial phagocytosis by macrophages. Maintenance of low c-di-GMP concentrations by these phosphodiesterases was required to promote survival within macrophages and virulence for mice. Attenuation of S Typhimurium virulence was due to overproduction of c-di-GMP-regulated cellulose, as deletion of the cellulose synthase machinery restored virulence to a strain lacking enzymatic activity of the three phosphodiesterases. We further identified that the cellulose-mediated reduction in survival was constrained to a slow-replicating persister population of S. Typhimurium induced within the macrophage intracellular environment. As utilization of glucose has been shown to be required for S. Typhimurium macrophage survival, one possible hypothesis is that this persister population requires the glucose redirected to the synthesis of cellulose to maintain a slow-replicating, metabolically active state.


Assuntos
GMP Cíclico/análogos & derivados , Citoplasma/metabolismo , Citoplasma/microbiologia , Macrófagos/metabolismo , Macrófagos/microbiologia , Salmonella typhimurium/patogenicidade , Animais , Técnicas Biossensoriais/métodos , Celulose/metabolismo , GMP Cíclico/metabolismo , Modelos Animais de Doenças , Fluorescência , Glucosiltransferases , Interações Hospedeiro-Patógeno/fisiologia , Camundongos , Camundongos Endogâmicos BALB C , Viabilidade Microbiana , Fagocitose , Diester Fosfórico Hidrolases/metabolismo , Salmonella typhimurium/metabolismo , Virulência
12.
Toxicon ; 163: 12-18, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30880186

RESUMO

Spider venoms are widely recognized as a new emerging source of potential research tools, pesticides, drug leads, and therapeutic agents. Some studies suggest that these venoms may contain interesting vasodilator compounds with potential therapeutic applications. In the present study, the vasodilator activity of the venom of Poecilotheria regalis was evaluated in isolated rat aortic rings. This venom induced an endothelium-dependent vasodilation [EC50 value was 5.52 (4.18-7.32) µg protein/ml with an Emax = 103.4 ±â€¯3.8%]. While the percentage of vasodilation induced by the venom was significantly diminished in the presence of a nitric oxide synthase inhibitor (L-NAME), it remained unaltered in the presence of suramin, a P2-purinergic receptor antagonist. Moreover, the vasodilator activity of the venom was not affected after boiling bath incubation, but was significantly decreased under reducing conditions. Additionally, venom composition was analyzed by reverse-phase chromatography and MALDI-TOF mass spectrometry, and two fractions were obtained, referred to as peptidic and non-peptidic fractions. Interestingly, both fractions induced vasodilation in isolated rat aortic rings. The results of this study showed that the venom of P. regalis induces a concentration-dependent vasodilation in rat aorta that was endothelium-dependent and involves the activation of NO/cGMP pathway. These results suggest that the venom contains a combination of both peptidic and non-peptidic vasodilator components. This study provides pharmacological data that suggest that P. regalis venom may be an important source of peptidic and non-peptidic vasodilator compounds.


Assuntos
Venenos de Aranha/farmacologia , Aranhas , Vasodilatação/efeitos dos fármacos , Animais , Aorta/efeitos dos fármacos , GMP Cíclico/metabolismo , Endotélio Vascular/efeitos dos fármacos , Feminino , Técnicas In Vitro , Masculino , Óxido Nítrico/metabolismo , Ratos , Ratos Wistar , Venenos de Aranha/química , Vasodilatadores/farmacologia
13.
Int J Mol Sci ; 20(6)2019 Mar 19.
Artigo em Inglês | MEDLINE | ID: mdl-30893788

RESUMO

Ultraviolet (UV) filters are chemicals widely used in personal care products (PCPs). Due to their effect as endocrine disruptor compounds (EDCs), the toxicity of UV filters is a current concern for human health. EDC exposure may be correlated to cardiovascular diseases (CVD), but to our knowledge, no studies assessed the UV filters effects as human EDCs at the vascular level. Octylmethoxycinnamate (OMC) is the world's most widely used UV-B filter, present in more than 90% of PCPs. Due to its demonstrated multiple hormonal activities in animal models, this substance is also suspected to be a human EDC. The purpose of this study was to assess the rapid/short-term effects of OMC on arterial tonus and analyse its mode of action (MOA). Using human umbilical arteries, the endocrine effects of OMC were evaluated in in vitro (cellular and organ) experiments by planar cell surface area (PCSA) and organ bath, respectively. Our data show that OMC induces a rapid/short-term smooth muscle relaxation acting through an endothelium-independent MOA, which seems to be shared with oestrogens, involving an activation of soluble guanylyl cyclase (sGC) that increases the cyclic guanosine monophosphate (cGMP) intracellular levels and an inhibition of L-type voltage-operated Ca2+ channels (L-Type VOCC).


Assuntos
Bloqueadores dos Canais de Cálcio/farmacologia , Cinamatos/farmacologia , Guanilil Ciclase Solúvel/metabolismo , Raios Ultravioleta , Artérias Umbilicais/enzimologia , Artérias Umbilicais/fisiologia , Vasodilatação/efeitos dos fármacos , GMP Cíclico/metabolismo , Ativação Enzimática/efeitos dos fármacos , Humanos , Modelos Biológicos , Miócitos de Músculo Liso/efeitos dos fármacos , Miócitos de Músculo Liso/metabolismo , Vasoconstrição/efeitos dos fármacos
14.
Ann Clin Lab Sci ; 49(1): 97-104, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30814084

RESUMO

PURPOSE: The objective of this research was to explore the effect of dioscin on myocardium in streptozotocin (STZ)-induced diabetic rats and the underlying mechanisms. METHODS: Diabetic rat model was established by a single intravenous injection of streptozocin (STZ). The rats were divided into 5 groups: control group, control+dioscin group, model group (diabetes), DDL group (diabetic rats treated with 100 µg/kg/day dioscin) and DDH group (diabetic rats treated with 200 µg/kg/day dioscin). Each group was continuously intervened for 6 weeks. Hemodynamic parameters were detected and pathological alterations of myocardium were observed by hematoxylin-eosin (HE) staining. Inflammatory response and related proteins in the NO-sGC-cGMP-PKG pathway were detected by western blot. RESULTS: Dioscin treatment can increase ejection fraction (EF) and decrease left ventricular end-diastolic pressure (LVEDP) as well as time constant of left ventricular pressure decay (Tau) parameters in diabetic rats, suggesting the improvement of left ventricular function. By histopathology observation, we found that dioscin treatment can also improve myocardial histological lesions caused by diabetes. In addition, the levels of inflammatory cytokines TGF-ß1, TNF-α and IL-1ß of the model group were remarkably higher than those in the control group (p<0.01), while after being treated with dioscin these cytokines were obviously decreased (p<0.05). The levels of PDE-5, PKG and p-VASP in the diabetic rats were significantly declined after being treated by dioscin in a dose-dependent manner (p<0.05). CONCLUSION: Dioscin may prevent the myocardial injury in diabetic rats by up-regulating NO-sGC-cGMP-PKG pathway.


Assuntos
Proteínas Quinases Dependentes de GMP Cíclico/metabolismo , GMP Cíclico/metabolismo , Diabetes Mellitus Experimental/complicações , Cardiomiopatias Diabéticas/prevenção & controle , Diosgenina/análogos & derivados , Guanilato Ciclase/metabolismo , Óxido Nítrico/metabolismo , Transdução de Sinais/efeitos dos fármacos , Animais , Diabetes Mellitus Experimental/fisiopatologia , Cardiomiopatias Diabéticas/etiologia , Cardiomiopatias Diabéticas/metabolismo , Diosgenina/farmacologia , Modelos Animais de Doenças , Fibrose/etiologia , Fibrose/metabolismo , Fibrose/prevenção & controle , Ratos , Ratos Sprague-Dawley
15.
Int J Med Sci ; 16(3): 443-449, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30911278

RESUMO

Background: Hypoxia plays an important role in the vascular tone of pulmonary circulation via the vasculature and parenchymal tissue. Endothelin-1 (ET-1), a potent vasoconstrictive peptide, plays a role in inflammation in mononuclear cells. Nitric oxide synthase (NOS), which generates nitric oxide (NO)/cyclic 3', 5'-monophosphate (cGMP), is coexpressed with ET-1 in many cell types. The aim of this study was to assess whether hypoxia induces the production of ET-1 and associated expression of NOS, NO/cGMP and chemokines in rat alveolar macrophages (AMs). Methods: NR8383 cells were cultured under hypoxic (1% oxygen) conditions for 0, 2, 4, 8 and 12 hours. Levels of ET-1, inducible NOS (iNOS), phosphorylated iNOS (p-iNOS), nitrite/nitrate (NOx), cGMP and monocyte chemoattractant protein-1 (MCP-1) were measured. Results: ET-1, p-iNOS, NOx, and cGMP increased significantly in AMs after 4 hours of hypoxia (p < 0.05). ET-1 and MCP-1 mRNA increased after 8 hours (p < 0.05). The protein expression of ET-1, MCP-1, and p-iNOS increased in a time-dependent manner, while iNOS expression decreased with time. Conclusions: The changes in ET-1, p-iNOS, and the NO/cGMP pathway in AMs may help elucidate the mechanisms in the hypoxic lung. Understanding changes in the endothelin axis in hypoxic AMs is a crucial first step to unravel its role in pulmonary circulation.


Assuntos
GMP Cíclico/metabolismo , Endotelina-1/metabolismo , Macrófagos Alveolares/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Animais , Hipóxia Celular , Células Cultivadas , Quimiocina CCL2/genética , Quimiocina CCL2/metabolismo , Endotelina-1/genética , Regulação da Expressão Gênica , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/genética , Ratos Sprague-Dawley
16.
Pak J Pharm Sci ; 32(1(Supplementary)): 261-268, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30829202

RESUMO

Using rat thoracic aortic rings to test the relaxing effects of the 95% ethanol extract and aqueous extract of Taohong Siwu decoction (THSW) on endothelium intact or endothelium removed aortic rings. Results showed that the 95% ethanol extract (0.1, 1, 10, 100, 1000 mg•L-1) and aqueous extract (0.1, 1, 10, 100, 1000 mg•L-1) of THSW were able to relax the intact endothelium aortic rings pre-contracted by 10-6 mol•L-1 PE. 10-4 mol•L-1 L-NAME and 10-5 mol•L-1 methylene blue both were able to inhibit the relaxation other than indomethacin. For the endothelium removed aortic rings, potassium channel blocker 3×10-3mol•L-1 tetraethylammonium chloride and 10-5 mol•L-1 glibenclamide had no effect on the relaxation effects caused by the 95% ethanol extract and aqueous extract of THSW. It could be concluded that the 95% ethanol extract and aqueous extract of THSW relax blood vessel by endothelium-dependent way.


Assuntos
Aorta/efeitos dos fármacos , GMP Cíclico/metabolismo , Medicamentos de Ervas Chinesas/farmacologia , Óxido Nítrico/metabolismo , Vasodilatadores/farmacologia , Animais , Endotélio Vascular/efeitos dos fármacos , Glibureto/farmacologia , NG-Nitroarginina Metil Éster/farmacologia , Técnicas de Cultura de Órgãos , Bloqueadores dos Canais de Potássio/farmacologia , Ratos , Tetraetilamônio/farmacologia
17.
Biosci Biotechnol Biochem ; 83(6): 1000-1010, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30835622

RESUMO

Pulmonary hypertension (PH) is a life-threatening lung disease. PH with concomitant lung diseases, e.g., idiopathic pulmonary fibrosis, is associated with poor prognosis. Development of novel therapeutic vasodilators for treatment of these patients is a key imperative. We evaluated the efficacy of dual activation of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) using an active, small-molecule phosphodiesterase (PDE4)/PDE5 dual inhibitor (Compound A). Compound A increased both cAMP and cGMP levels in WI-38 lung fibroblasts and suppressed the expressions of type-1 collagen α1 chain and fibronectin. Additionally, compound A reduced right ventricular weight/left ventricular weight+septal weight ratio, brain natriuretic peptide expression levels in right ventricle, C─C motif chemokine ligand 2 expression levels in lung, and plasma surfactant protein D. Our data indicate that dual activation of cAMP/cGMP pathways may be a novel treatment strategy for PH.


Assuntos
AMP Cíclico/metabolismo , GMP Cíclico/metabolismo , Hipertensão Pulmonar/induzido quimicamente , Hipertensão Pulmonar/tratamento farmacológico , Inflamação/terapia , Pulmão/efeitos dos fármacos , Monocrotalina/toxicidade , Inibidores da Fosfodiesterase 5/uso terapêutico , Animais , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Células Cultivadas , Colágeno Tipo I/metabolismo , Modelos Animais de Doenças , Epitélio/lesões , Fibronectinas/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/fisiologia , Humanos , Pulmão/metabolismo , Pulmão/patologia , Inibidores da Fosfodiesterase 5/farmacologia , Ratos Wistar , Fator de Crescimento Transformador beta/fisiologia
18.
Biomed Pharmacother ; 111: 1243-1248, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30841438

RESUMO

AIM: To observe the effect of propranolol in cervical cancer and investigate the mechanism of the effect. METHODS AND RESULTS: We found 5 direct protein targets (DPTs) of propranolol (PRO) by DrugBank5.0 firstly. Next, we analyzed protein-protein interaction (PPI) network and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways of PRO DPTs and the result showed that PRO was linked with cGMP/PKG pathway. Then, we recognized the top 38 upexpressed genes of cervical cancer (CC) based original microarray datasets (GSE7803, GSE9750, GSE39001 and GSE63514). Further, we analyzed the biological process with the 38 overexpressed genes by STRING. We found some of overexpressed genes of CC participated in GMP biosynthetic process. Lastly, the function of PRO in CC was validated by MTT assay, Western blotting, flow cytometry and colony formation assay methods. We verified PRO can suppress cGMP/PKG pathway then inhibits CC cell growth. CONCLUSION: The bioinformatical analysis combine with traditional experiment can help us understanding potential molecular mechanism about how PRO acting in CC. This method is a new paradigm which can guide future researches about mechanism in existing diseases and drugs.


Assuntos
Proliferação de Células/efeitos dos fármacos , Proteínas Quinases Dependentes de GMP Cíclico/metabolismo , GMP Cíclico/metabolismo , Propranolol/farmacologia , Transdução de Sinais/efeitos dos fármacos , Neoplasias do Colo do Útero/tratamento farmacológico , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Feminino , Células HeLa , Humanos , Mapas de Interação de Proteínas/efeitos dos fármacos , Neoplasias do Colo do Útero/metabolismo
19.
Biomed Pharmacother ; 111: 1458-1466, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30841461

RESUMO

INTRODUCTION: Prostaglandins (PGs) play an important role in corpus cavernosum relaxation, as evidenced by alprostadil being used as a drug for erectile dysfunction. Reports about the effect of cyclooxygenase (COX) inhibitors on erectile function are highly contradictory. AIM: To compare the potential effects of some COX inhibitors with varying COX-1/COX-2 selectivities (indomethacin, ketoprofen and diclofenac) with that of the selective COX-2 inhibitor (DFU) on corpus cavernosal tone in-vitro. The role played by PGE1, PGI2-analogue and PGE4 receptor (EP4)-agonist in controlling corpus cavernosum function and the modulation of their action by sildenafil is also studied. METHODS: Organ bath experiments were performed using isolated rat corpus cavernosum. Direct relaxations and changes to electric field stimulation (EFS, 2-16 Hz, 60 V, 0.8 ms, 10 s train)-induced relaxation by the effect of the selected drugs were studied. Strips were precontracted using phenylephrine (PE, 10-5 M). Results are expressed as mean ± SEM of 5-9 rats. RESULTS: Alprostadil, iloprost and L902688 (selective EP4 agonist) induced direct relaxation where L902688 showed greater relaxant effect. Sildenafil potentiated the Emax of alprostadil and iloprost but not L902688. EFS and acetylcholine (ACh)-induced relaxations were significantly potentiated in presence of indomethacin, ketoprofen and diclofenac (20, 100 µM) but not in presence of selective COX-2 inhibitor (DFU, 1 µM). GR32191B (Thromboxane A2 receptor antagonist, 10-6 M) significantly reduced the potentiatory effect of indomethacin. Only diclofenac succeeded to potentiate sodium nitroprusside (SNP)-induced relaxation. CONCLUSIONS: EP4 receptors may play an important nitric oxide (NO)/cGMP-independent role in corpus cavernosal relaxation. Nonselective COX inhibitors seem of no harm concerning cavernosal tissue relaxation, possibly because they inhibit the synthesis of the highly contracting mediator thromboxane A2.


Assuntos
Inibidores de Ciclo-Oxigenase/farmacologia , Relaxamento Muscular/efeitos dos fármacos , Pênis/efeitos dos fármacos , Prostaglandinas/metabolismo , Alprostadil/farmacologia , Animais , GMP Cíclico/metabolismo , Diclofenaco/farmacologia , Disfunção Erétil/tratamento farmacológico , Disfunção Erétil/metabolismo , Iloprosta/farmacologia , Indometacina/farmacologia , Cetoprofeno/farmacologia , Masculino , Óxido Nítrico/metabolismo , Nitroprussiato/farmacologia , Ereção Peniana/efeitos dos fármacos , Pênis/metabolismo , Piperazinas/farmacologia , Ratos , Citrato de Sildenafila/farmacologia , Sulfonas/farmacologia
20.
Molecules ; 24(5)2019 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-30862086

RESUMO

Recently, our research group demonstrated that uvaol and ursolic acid increase NO and H2S production in aortic tissue. Molecular docking studies showed that both compounds bind with high affinity to endothelial NO synthase (eNOS) and cystathionine gamma-lyase (CSE). The aim of this study was to identify hits with high binding affinity for the triterpene binding-allosteric sites of eNOS and CSE and to evaluate their vasodilator effect. Additionally, the mechanism of action of the most potent compound was explored. A high-throughput virtual screening (HTVS) of 107,373 compounds, obtained from four ZINC database libraries, was performed employing the crystallographic structures of eNOS and CSE. Among the nine top-scoring ligands, isoxsuprine showed the most potent vasodilator effect. Pharmacological evaluation, employing the rat aorta model, indicated that the vasodilation produced by this compound involved activation of the NO/cGMP and H2S/KATP signaling pathways and blockade of α1-adrenoceptors and L-type voltage-dependent Ca2+ channels. Incubation of aorta homogenates in the presence of isoxsuprine caused 2-fold greater levels of H2S, which supported our preliminary in silico data. This study provides evidence to propose that the vasodilator effect of isoxsuprine involves various mechanisms, which highlights its potential to treat a wide variety of cardiovascular diseases.


Assuntos
Antagonistas de Receptores Adrenérgicos alfa 1/farmacologia , Bloqueadores dos Canais de Cálcio/farmacologia , Isoxsuprina/química , Isoxsuprina/farmacologia , Redes e Vias Metabólicas/efeitos dos fármacos , Relação Quantitativa Estrutura-Atividade , Vasodilatadores/química , Vasodilatadores/farmacologia , Trifosfato de Adenosina/metabolismo , Antagonistas de Receptores Adrenérgicos alfa 1/química , Bloqueadores dos Canais de Cálcio/química , GMP Cíclico/metabolismo , Relação Dose-Resposta a Droga , Ensaios de Triagem em Larga Escala , Humanos , Sulfeto de Hidrogênio/metabolismo , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Óxido Nítrico/metabolismo , Bibliotecas de Moléculas Pequenas , Fluxo de Trabalho
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