RESUMO
Solidagenone (SOL) is a labdane-type diterpenoid found in Solidago chilensis, a plant traditionally used to treat skin diseases, kidney pain and ovarian inflammation. In this study, the topical anti-inflammatory activity of SOL was evaluated using in vivo and in silico assays. Croton oil-, arachidonic acid (AA)- and phenol-induced ear oedema mouse models were applied in the in vivo studies. Myeloperoxidase (MPO) and N-acetyl-ß-D-glucosaminidase (NAG) activities and tumour necrosis factor alpha (TNF-α), interleukin-6 (IL-6) and nitric oxide (NO) levels were determined, as well as histopathological analyses were conducted. Interaction profiles between SOL and cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), glucocorticoid receptor, estradiol-17-ß-dehydrogenase and prostaglandin-E(2)-9-reductase were established using molecular docking. SOL significantly inhibited croton oil-, AA- and phenol-induced ear oedema (P < .001) at doses of 0.1, 0.5 and 1.0 mg/ear. The MPO and NAG activities and TNF-α, IL-6 and NO levels were decreased (P < .001). The histopathological data revealed that inflammatory parameters (oedema thickness, leucocyte infiltration and vasodilatation) were reduced by treatment with SOL at doses of 0.1, 0.5 and 1.0 mg/ear. The docking study showed that SOL interacts with COX-1 and prostaglandin-E(2)-9-reductase through hydrogen bonding, inhibiting these enzymes. These results indicate that SOL may be a promising compound for the treatment of cutaneous inflammatory disorders and has potential as a topical anti-inflammatory agent.
Assuntos
Inibidores de Ciclo-Oxigenase/farmacologia , Dermatite/prevenção & controle , Edema/prevenção & controle , Furanos/farmacologia , Hidroxiprostaglandina Desidrogenases/antagonistas & inibidores , Proteínas de Membrana/antagonistas & inibidores , Naftalenos/farmacologia , Extratos Vegetais/farmacologia , Pele/efeitos dos fármacos , Solidago , Acetilglucosaminidase/metabolismo , Animais , Ciclo-Oxigenase 1/metabolismo , Inibidores de Ciclo-Oxigenase/isolamento & purificação , Inibidores de Ciclo-Oxigenase/metabolismo , Dermatite/metabolismo , Dermatite/patologia , Modelos Animais de Doenças , Edema/induzido quimicamente , Edema/metabolismo , Edema/patologia , Furanos/isolamento & purificação , Furanos/metabolismo , Ligação de Hidrogênio , Hidroxiprostaglandina Desidrogenases/metabolismo , Interleucina-6/metabolismo , Masculino , Proteínas de Membrana/metabolismo , Camundongos , Simulação de Acoplamento Molecular , Naftalenos/isolamento & purificação , Naftalenos/metabolismo , Óxido Nítrico/metabolismo , Peroxidase/metabolismo , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/metabolismo , Ligação Proteica , Transdução de Sinais , Pele/metabolismo , Pele/patologia , Solidago/química , Fator de Necrose Tumoral alfa/metabolismoRESUMO
The initial inactivation of prostaglandins (PGs) is mediated by 15-hydroxyprostaglandin dehydrogenase (15-PGDH). PGs are potent mediators of several biological processes, including inflammation and reproduction. In uterus, PGs play a key role in infection-induced pregnancy loss, in which concentration of this mediator increased. This process is accompanied with the induction of nitric oxide synthase expression and a marked increase in uterine levels of nitric oxide. There is no information concerning nitric oxide contribution to potential changes in PG catabolism, but experimental evidence suggests that nitric oxide modulates PG pathways. The specific objectives of the study were to evaluate the protein expression of HPGD (15-PGDH) and to characterize the nitric oxide-dependent regulation of this enzyme in a model of lipopolysaccharide (LPS)-induced embryonic resorption. Results show that LPS decreased HPGD protein expression and augmented PGE synthase activity; therefore, PGE2 levels increased in uterus in this inflammatory condition. Just as LPS, the treatment with a nitric oxide donor diminished HPGD protein expression in uterine tissue. In contrast, the inhibition of nitric oxide synthesis both in control and in LPS-treated mice increased 15-PGDH levels. Also, we have found that this enzyme and PGE2 levels are not modulated by peroxynitrite, an oxidant agent derived from nitric oxide. This study suggests that LPS and nitric oxide promote a decrease in the ability of the uterus for PG catabolism during bacterially triggered pregnancy loss in mice.
Assuntos
Regulação para Baixo , Perda do Embrião/metabolismo , Hidroxiprostaglandina Desidrogenases/metabolismo , Óxido Nítrico/metabolismo , Útero/metabolismo , Animais , Dinoprostona/metabolismo , Regulação para Baixo/efeitos dos fármacos , Perda do Embrião/enzimologia , Perda do Embrião/imunologia , Inibidores Enzimáticos/farmacologia , Infecções por Escherichia coli/enzimologia , Infecções por Escherichia coli/imunologia , Infecções por Escherichia coli/metabolismo , Feminino , Hidroxiprostaglandina Desidrogenases/antagonistas & inibidores , Oxirredutases Intramoleculares/metabolismo , Lipopolissacarídeos , Camundongos , Camundongos Endogâmicos BALB C , Doadores de Óxido Nítrico/farmacologia , Óxido Nítrico Sintase/antagonistas & inibidores , Gravidez , Complicações Infecciosas na Gravidez/enzimologia , Complicações Infecciosas na Gravidez/imunologia , Complicações Infecciosas na Gravidez/metabolismo , Prostaglandina-E Sintases , Distribuição Aleatória , Regulação para Cima/efeitos dos fármacos , Útero/efeitos dos fármacos , Útero/imunologiaRESUMO
We studied (1) the effects of oral contraceptive pills (OCPs) on hirsutism, hormonal and metabolic variables in 49 polycystic ovary syndrome patients without metabolic comorbidities and (2) the effect of 17-hydroxysteroid dehydrogenase type 5 gene polymorphism (-71A/G HSD17B5 SNP) on the response to OCP treatment. Mean age was 21.9 ± 6.5 years. Patients received monophasic OCP (20 µg ethinyl estradiol plus 75 µg gestodene), 21/28 days per cycle, during 6 months; 32 patients with severe hirsutism also received spironolactone 100 mg. The frequencies of HSD17B5 genotypes were: AA = 0.49 (55.1%), AG = 0.42 (30.6%) and GG = 0.09 (14.3%). After 6 months, body mass index and waist circumference remained unchanged regardless of the presence of allele G. A slight reduction (p < 0.05) was noted in systolic blood pressure (p < 0.05) and luteinizing hormone levels, whereas a slight increase (p < 0.05) was noted in lipids. Total testosterone and hirsutism score declined, while sex hormone binding globulin increased after OCP treatment (p < 0.05). None of these changes were associated with genotype. Insulin and homeostasis model assessment remained unchanged after treatment and did not vary according to the presence of allele G. OCP seems to ameliorate androgenic symptoms without compromising metabolic parameters. The -71A/G SNP of HSD17B5 gene did not contribute to the improvements observed.
Assuntos
Anticoncepcionais Orais Combinados/uso terapêutico , Etinilestradiol/uso terapêutico , Hirsutismo/prevenção & controle , Norpregnenos/uso terapêutico , Síndrome do Ovário Policístico/tratamento farmacológico , Globulina de Ligação a Hormônio Sexual/análise , Testosterona/sangue , 3-Hidroxiesteroide Desidrogenases/genética , 3-Hidroxiesteroide Desidrogenases/metabolismo , Adolescente , Adulto , Membro C3 da Família 1 de alfa-Ceto Redutase , Brasil , Anticoncepcionais Orais Combinados/efeitos adversos , Quimioterapia Combinada , Etinilestradiol/efeitos adversos , Feminino , Estudos de Associação Genética , Hirsutismo/tratamento farmacológico , Hirsutismo/etiologia , Hirsutismo/fisiopatologia , Humanos , Hidroxiprostaglandina Desidrogenases/genética , Hidroxiprostaglandina Desidrogenases/metabolismo , Hiperandrogenismo/etiologia , Hiperandrogenismo/prevenção & controle , Antagonistas de Receptores de Mineralocorticoides/uso terapêutico , Norpregnenos/efeitos adversos , Projetos Piloto , Síndrome do Ovário Policístico/genética , Síndrome do Ovário Policístico/metabolismo , Síndrome do Ovário Policístico/fisiopatologia , Polimorfismo de Nucleotídeo Único , Índice de Gravidade de Doença , Espironolactona/uso terapêutico , Adulto JovemRESUMO
The first proteomic analysis of Trypanosoma cruzi resistance to Benznidazole (BZ) is presented. The differential proteome of T. cruzi with selected in vivo resistance to Benznidazole (BZR and Clone27R), its susceptible pairs (BZS and Clone9S), and a pair from a population with Benznidazole- in vitro-induced resistance (17LER) and the susceptible pair 17WTS were analyzed by two-dimensional gel electrophoresis (2-DE) followed by mass spectrometry (MS) for protein identification. Out of 137 spots analyzed through MS, 110 were identified as 56 distinct proteins. Out of the 56 distinct proteins, 36 were present in resistant, 9 in susceptible, and 11 in both phenotypes. Among the proteins identified in resistant samples, 5 were found in Cl 27R and in BZR (calpain-like cysteine peptidase, hypothetical protein conserved 26 kDa, putative peptidase, peroxiredoxin and tyrosine amino transferase) and 4 in Cl 27R and 17LER (cyclophilin A, glutamate dehydrogenase, iron superoxide dismutase and nucleoside diphosphate kinase). As for the proteins identified in Benznidazole-susceptible samples, PGF-2a was found in BZS and 17WTS. A functional category analysis showed that the proteins involved with transcription and protein destination were overexpressed for the Benznidazole-resistant phenotype. Thus, the present study provides large-scale, protein-related information for investigation of the mechanism of T. cruzi resistance to Benznidazole.
Assuntos
Resistência a Medicamentos , Nitroimidazóis/farmacologia , Proteoma/metabolismo , Proteínas de Protozoários/metabolismo , Trypanosoma cruzi/metabolismo , Animais , Ciclofilina A/análise , Ciclofilina A/metabolismo , Cisteína Endopeptidases/análise , Cisteína Endopeptidases/metabolismo , Eletroforese em Gel Bidimensional , Glutamato Desidrogenase/análise , Glutamato Desidrogenase/metabolismo , Hidroxiprostaglandina Desidrogenases/análise , Hidroxiprostaglandina Desidrogenases/metabolismo , Núcleosídeo-Difosfato Quinase/análise , Núcleosídeo-Difosfato Quinase/metabolismo , Peptídeo Hidrolases/análise , Peptídeo Hidrolases/metabolismo , Peroxirredoxinas/análise , Peroxirredoxinas/metabolismo , Proteoma/análise , Proteínas de Protozoários/análise , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Superóxido Dismutase/análise , Superóxido Dismutase/metabolismo , Espectrometria de Massas em Tandem , Tripanossomicidas/farmacologia , Trypanosoma cruzi/efeitos dos fármacos , Tirosina Transaminase/análise , Tirosina Transaminase/metabolismoRESUMO
Nitric oxide (NO) synthesized by fetal membranes may act either directly inhibiting myometrium contractility or indirectly interacting with tocolytic agents as prostaglandins (PGs). Here we examined if NO could modulate prostaglandin E(2) 9-ketoreductase (9-KPR) activity in human fetal membranes (HFM). 9-KPR is the enzyme that converts PGE(2) into PGF(2alpha), the main PGs known to induce uterine contractility at term. Chorioamnion explants obtained from elective caesareans were incubated with aminoguanidine (AG), an iNOS inhibitor, or NOC-18, a NO donor. NOC-18 (2mM) increased PGE(2) production and diminished PGF(2alpha) synthesis in HFM. AG presented the opposite effect. When we evaluated the activity of 9-KPR by the conversion of [(3)H]-PGE(2) into [(3)H]-PGF(2alpha) and 13,14-dihidro-15-keto prostaglandin F(2alpha) (the PGF(2alpha) metabolite), we found that NOC-18 inhibited 9-KPR activity. Interestingly, AG did not elicit any effect on 9-KPR but l-NAME, a non-selective NOS inhibitor, significantly increased its activity. Our data suggests that exogenous NO inhibits 9-KPR activity in HFM, thus modulating the synthesis of important labor mediators as PGF(2alpha).
Assuntos
Membranas Extraembrionárias/enzimologia , Hidroxiprostaglandina Desidrogenases/metabolismo , Óxido Nítrico/metabolismo , Dinoprosta/metabolismo , Dinoprostona/metabolismo , Relação Dose-Resposta a Droga , Membranas Extraembrionárias/citologia , Membranas Extraembrionárias/efeitos dos fármacos , Feminino , Regulação da Expressão Gênica , Humanos , Hidroxiprostaglandina Desidrogenases/antagonistas & inibidores , Pessoa de Meia-Idade , Doadores de Óxido Nítrico/farmacologia , Óxido Nítrico Sintase/metabolismo , Gravidez , Prostaglandina-Endoperóxido Sintases/metabolismoRESUMO
Myometrial quiescence is a key factor in all species to accomplish a successful gestation. PGs play a crucial role in mediating parturition events, and their synthesis and metabolism are regulated by cyclooxygenases (COXs) and NAD(+)-dependent 15-hydroxy-PG dehydrogenase (PGDH), respectively. Progesterone (P(4)) is the hormone responsible for maintaining uterine smooth muscle quiescence during pregnancy. In this work, we have studied the effect of P(4) on the activity of COXs and PGDH, the uterine enzymes involved in the biosynthesis and metabolism of prostanoids in the rat. We found that during pregnancy PGF(2alpha) production and also protein levels of COX-1 and COX-2 were decreased. The exogenous administration of P(4) significantly inhibited the uterine production of PGF(2alpha) and also the protein level of COX-2. PGF(2alpha), metabolism was assessed by PGDH activity, which resulted high during pregnancy and increased as a result of P(4) administration. These results indicate that PGs levels were negatively modulated by P(4), which could be exerting its effect by increasing PGs metabolism through stimulation on PGDH activity and an inhibition on COX and that is a major mechanism for maintain uterine quiescence in pregnancy.
Assuntos
Abortivos não Esteroides/metabolismo , Dinoprosta/metabolismo , Progesterona/farmacologia , Útero/efeitos dos fármacos , Animais , Ciclo-Oxigenase 1 , Ciclo-Oxigenase 2 , Feminino , Hidroxiprostaglandina Desidrogenases/metabolismo , Isoenzimas/metabolismo , Proteínas de Membrana , Gravidez , Prostaglandina-Endoperóxido Sintases/metabolismo , Ratos , Ratos Wistar , Útero/metabolismoRESUMO
The effects of exogenous histamine (H) on prostaglandin (PG) generation and release in uteri isolated from diestrous rats and the influences of H2-receptors blockers (cimetidine and metiamide) on the output of uterine PGs, were explored. Moreover, the action of H on the uterine 9-keto-reductase, was also studied. Histamine (10(-4) M) failed to alter the basal output of PGE1 but reduced significantly the generation and release of PGE2 and augmented the output of PGF2 alpha. On the other hand, cimetidine (10(-5) M) enhanced the basal release of PGE2 but had no action on the outputs of PGs E1 or F2 alpha. The enhancing effect of H on the production and release of PGF2 alpha was abolished in the presence of cimetidine. Also, the antagonist reversed the influence of H on the output of PGE2. Metiamide, another H2-receptor antagonist, did not alter the basal control generation and release of uterine PGs, but antagonized the augmenting influence of H on PGF2 alpha uterine output, as much as cimetidine did, and prevented the depressive action of H on the release of PGE2 from uteri. Histamine (10(-4) M) significantly stimulated uterine formation of cyclic-adenosine monophosphate, an action which was antagonized by the presence of cimetidine (10(-5) M), a blocker of H2 receptors. Also, histamine (10(-5) M) and dibutyrylcyclic-adenosine monophosphate (DB-cAMP) at 10(-3) M, enhanced significantly the formation 3H-PGF2 alpha from 3H-PGE2. Results presented herein demonstrate that H is able to diminish the generation of PGE2 in uteri from rats at diestrus augmenting the synthesis of PGF2 alpha, apparently via the activation of H2-receptors, enhancing adenylate-cyclase. These effects appear to increase uterine 9-keto-reductase activity which transforms PGE2 into PGF2 alpha. Relationships between the foregoing results and those evoked by estradiol, are also discussed.
Assuntos
Diestro/metabolismo , Estro/metabolismo , Histamina/farmacologia , Hidroxiprostaglandina Desidrogenases/metabolismo , Prostaglandinas/biossíntese , Receptores Histamínicos H2/efeitos dos fármacos , Útero/metabolismo , Animais , Cimetidina/farmacologia , AMP Cíclico/metabolismo , Estradiol/metabolismo , Feminino , Técnicas In Vitro , Metiamida/farmacologia , Ratos , Ratos EndogâmicosRESUMO
The generation and output of prostaglandins (PGs) E2 and F2 alpha into the solution suspending uterine segments from ethanol (ETOH)-fed diestrous rats and the activity of 15-OH-PG-dehydrogenase (PGDH) in uteri at diestrus, were explored and compared with normal-fed controls. Animals were fed with ETOH (35% of the total calories in a liquid diet) during 20 days before sacrifice. Paired normal-fed controls were given isocaloric quantities of dextrimaltose. It was observed that the uterine outputs of PGE2 and of PGF2 alpha into the suspending solution, were significantly greater in the ETOH group. On the other hand, the PGDH activity for PGE2 in control uterine tissue, was significantly smaller than the activity detected in preparations from animals fed with the chronic ETOH diet. Results are discussed in terms of possible mechanisms for the action of ethanol, either on the release of PG fatty acid precursors (activation of phospholipase A2) or on the activity of PG synthesizing enzymes. Inasmuch as in the ETOH-fed group uterine PGDH activity was greater, rather than diminished, the possibility of a reduced catabolism accounting for the augmentation of PGs in the suspending medium, does not appear feasible. In fact, results suggest that the real magnitude of higher PG generation and release is even greater than that disclosed by the present study. The finding that chronic ethanol consumption augments PG production, appears relevant, in view of the unique roles played by these eicosanoids in parturition and in the development of fetuses.