RESUMO
The traditionally used oriental herbal medicine Moutan Cortex Radicis [MCR; Paeonia Suffruticosa Andrews (Paeoniaceae)] exerts anti-inflammatory, anti-spasmodic, and analgesic effects. In the present study, we investigated the therapeutic effects of differently fractioned MCR extracts in a 6-hydroxydopamine (OHDA)-induced Parkinson's disease model and neuro-blastoma B65 cells. Ethanol-extracted MCR was fractionated by n-hexane, butanol, and distilled water. Adult Sprague-Dawley rats were treated first with 20 µg of 6-OHDA, followed by three MCR extract fractions (100 or 200 mg·kg-1) for 14 consecutive days. In the behavioral rotation experiment, the MCR extract-treated groups showed significantly decreased number of net turns compared with the 6-OHDA control group. The three fractions also significantly inhibited the reduction in tyrosine hydroxylase-positive cells in the substantia nigra pars compacta following 6-OHDA neurotoxicity. Western blotting analysis revealed significantly reduced tyrosine hydroxylase expression in the substantia nigra pars compacta in the 6-OHDA-treated group, which was significantly inhibited by the n-hexane or distilled water fractions of MCR. B65 cells were exposed to the extract fractions for 24 h prior to addition of 6-OHDA for 30 min; treatment with n-hexane or distilled water fractions of MCR reduced apoptotic cell death induced by 6-OHDA neurotoxicity and inhibited nitric oxide production and neuronal nitric oxide synthase expression. These results showed that n-hexane- and distilled water-fractioned MCR extracts inhibited 6-OHDA-induced neurotoxicity by suppressing nitric oxide production and neuronal nitric oxide synthase activity, suggesting that MCR extracts could serve as a novel candidate treatment for the patients with Parkinson's disease.
Assuntos
Antiparkinsonianos/uso terapêutico , Medicamentos de Ervas Chinesas/química , Oxidopamina/toxicidade , Paeonia/química , Transtornos Parkinsonianos/tratamento farmacológico , Fitoterapia , Extratos Vegetais/uso terapêutico , Animais , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Antiparkinsonianos/farmacologia , Morte Celular/efeitos dos fármacos , Linhagem Celular , Modelos Animais de Doenças , Neurônios/patologia , Óxido Nítrico/análise , Óxido Nítrico Sintase Tipo I/biossíntese , Transtornos Parkinsonianos/induzido quimicamente , Extratos Vegetais/farmacologia , Plantas Medicinais , Ratos , Ratos Sprague-Dawley , Substância Negra/efeitos dos fármacos , Substância Negra/enzimologia , Tirosina 3-Mono-Oxigenase/genética , Tirosina 3-Mono-Oxigenase/metabolismoRESUMO
OBJECTIVE: This study aimed to detect the effect of a new herbal extract salvianolic acid A (SalA) on gastrointestinal complications in diabetic rats. METHODS: Altogether 80 rats were divided randomly into five groups, including normal control (NC) group, high-fat (HF) diet group, diabetes mellitus (DM) control group, and DM treated with SalA (0.1 mg/kg and 0.3 mg/kg) groups, respectively. DM was induced by feeding the rats with HF diet and the administration of streptozotocin (30 mg/kg). Four weeks after the establishment of the DM model, the rats received SalA or double distilled water for 8 weeks. After the evaluation of intestinal motility, the animals were sacrificed and their intestines were isolated and collected. The levels of advanced glycation end-products (AGE) and malondialdehyde (MDA) were detected. Protein gene product 9.5 (PGP9.5) and neuronal nitric oxide synthase (nNOS) expressions in the intestine were also detected. RESULTS: Compared with the NC and HF rats, the DM control rats showed significantly increased blood glucose level and decreased weight. Compared with the DM control group, SalA did not influence their weight and blood glucose level, but significantly reduced the levels of AGE and MDA. Intestinal transit was promoted by SalA in diabetic rats, and the expressions of PGP9.5 and nNOS in the intestine were both upregulated. CONCLUSION: The effect of SalA on the intestinal motility of diabetic rats might be due to its antioxidant capacity and restoring nNOS expression.
Assuntos
Antioxidantes/farmacologia , Ácidos Cafeicos/farmacologia , Diabetes Mellitus Experimental/tratamento farmacológico , Motilidade Gastrointestinal/efeitos dos fármacos , Lactatos/farmacologia , Óxido Nítrico Sintase Tipo I/biossíntese , Animais , Antioxidantes/uso terapêutico , Ácidos Cafeicos/uso terapêutico , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/fisiopatologia , Avaliação Pré-Clínica de Medicamentos/métodos , Duodeno/metabolismo , Motilidade Gastrointestinal/fisiologia , Jejuno/metabolismo , Lactatos/uso terapêutico , Masculino , Músculo Liso/metabolismo , Ratos , Ratos Sprague-Dawley , Ubiquitina Tiolesterase/biossíntese , Regulação para Cima/efeitos dos fármacosRESUMO
BACKGROUND AND OBJECTIVE: Vasoconstriction and hypersensitivity to the vasoconstrictive action of serotonin occurs in the early stage of atherosclerosis. Vascular neural nitric oxide synthase (nNOS) plays an important role in the regulation of vascular tone and is vasoprotective against atherosclerosis. In this study, we intended to investigate the possible role of nNOS in mediating the effect of Chinese medicine Tongxinluo (TXL) to attenuate vasoconstriction induced by the chronic injury in the collared carotid artery. METHODS: Twenty-four male Wistar Kyoto rats were assigned to 2 treatments (n = 12): vehicle and TXL (400 mg·kg·d). After 2 weeks of treatment, adventitia injury was induced by placing a silicone collar around the left carotid artery for 2 weeks. Blood flow and vascular reactivity to serotonin were determined, and carotid arteries were harvested for morphometry, RT-PCR, and Western blotting analysis. Expression of nNOS and phosphorylated ERK1/2 was also analyzed in primary cultured vascular smooth muscle cells after TXL and/or ERK kinase inhibitor treatment. RESULTS: Adventitia injury induced by the placement of a silicone collar around the carotid artery for 2 weeks led to chronic vasoconstriction and vascular hypersensitivity to serotonin, which was attenuated by TXL treatment. TXL improved the carotid blood flow and normalized the vascular hypersensitivity to serotonin in collared carotid arteries. The expression of nNOS and phosphorylated ERK1/2 was increased by TXL treatment in both collared carotid artery and vascular smooth muscle cells indicating a possible contribution of ERK1/2 and nNOS signaling to the beneficial effects of TXL. Moreover, we showed that the effect of TXL to increase nNOS expression was mediated by the phosphorylated ERK1/2 since the effect could be abolished by the ERK kinase inhibitor PD98059. CONCLUSIONS: TXL increases nNOS expression in the collared carotid artery through activation of ERK1/2 signaling, which may have contributed to the attenuation of vasoconstriction induced by the collar-induced adventitia injury.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Regulação Enzimológica da Expressão Gênica , Sistema de Sinalização das MAP Quinases/fisiologia , Óxido Nítrico Sintase Tipo I/biossíntese , Vasoconstrição/fisiologia , Animais , Células Cultivadas , Ativação Enzimática/efeitos dos fármacos , Ativação Enzimática/fisiologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Masculino , Ratos , Ratos Endogâmicos WKY , Vasoconstrição/efeitos dos fármacosRESUMO
We have previously shown that intrathecal administration of the adrenomedullin (AM) receptor antagonist AM(22-52) produces a long-lasting anti-hyperalgesia effect. This study examined the hypothesis that AM recruits other pronociceptive mediators in complete Freund's adjuvant (CFA)-induced inflammation. Injection of CFA in the hindpaw of rat produced an increase in the expression of nNOS in dorsal root ganglion (DRG) and the spinal dorsal horn. An intrathecal administration of AM(22-52), but not the CGRP antagonist BIBN4096BS, abolished the CFA-induced increase of nNOS. Moreover, AM-induced increase of CGRP was inhibited by the nNOS inhibitors L-NAME and 7-nitroindazole in cultured ganglion explants. Addition of AM to ganglion cultures induced an increase in nNOS protein, which was attenuated by the PKA inhibitor H-89. Treatment with AM also concentration-dependently increased cAMP content and pPKA protein level, but not its non-phosphorylated form, in cultured ganglia. In addition, nNOS was shown to be co-localized with the AM receptor components calcitonin receptor-like receptor and receptor activity-modifying protein 2- and 3 in DRG neurons. The present study suggests that the enhanced activity of nitric oxide (NO) mediates the biological action of AM at the spinal level and that AM recruits NO-CGRP via cAMP/PKA signaling in a mechanistic pathway underlying CFA-induced hyperalgesia.
Assuntos
Adrenomedulina/farmacologia , Peptídeo Relacionado com Gene de Calcitonina/biossíntese , Proteínas Quinases Dependentes de AMP Cíclico/fisiologia , Adjuvante de Freund/farmacologia , Óxido Nítrico Sintase Tipo I/biossíntese , Transdução de Sinais/fisiologia , Animais , Western Blotting , Proteínas Quinases Dependentes de AMP Cíclico/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Ensaio de Imunoadsorção Enzimática , Gânglios Espinais/metabolismo , Imuno-Histoquímica , Indazóis/farmacologia , Isoquinolinas/farmacologia , Masculino , NG-Nitroarginina Metil Éster/farmacologia , Óxido Nítrico/fisiologia , Óxido Nítrico Sintase Tipo I/antagonistas & inibidores , Técnicas de Cultura de Órgãos , Inibidores de Proteínas Quinases/farmacologia , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Sulfonamidas/farmacologia , Gânglio Trigeminal/metabolismo , Regulação para CimaRESUMO
BACKGROUND: Intracavernous injection of cultured adipose-derived stem cells (ADSCs) effectively restores erectile function in cavernous nerve (CN)-injured rats when administered at the time of injury. However, culturing exposes ADSCs to the risk of contamination and dedifferentiation. OBJECTIVE: Explore the effect of uncultured autologous adipose-derived stromal vascular fraction (SVF) on improving erectile function in a rat model of CN injury when administered at the time of injury or 4 wk after injury. DESIGN, SETTING, AND PARTICIPANTS: Eighty-nine male Sprague Dawley rats were randomly divided into four groups. CN injury or sham surgery was performed at the start of the study, and rats were treated with either SVF or vehicle. Functional testing and histologic analysis were performed 12 wk after CN crush or sham surgery. INTERVENTION: We used intracavernous injection of saline immediately after CN crush (n=23), intracavernous injection of SVF immediately after CN crush (n=17), intracavernous injection of SVF 4 wk after CN crush (n=23), or sham surgery (n=26). OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: We studied intracavernous pressure (ICP) response to CN electrostimulation and performed histologic examination of midpenile cross-sections. Data were analyzed using one-way analysis of variance followed by the Tukey-Kramer test. RESULTS AND LIMITATIONS: Both immediate and delayed treatment with SVF resulted in a significantly increased ICP-to-mean arterial pressure ratio compared with the vehicle-treated group. Both immediate and delayed treatment with SVF significantly increased expression of neuronal nitric oxide synthase and neurofilament in dorsal penile nerves compared to the vehicle group. Furthermore, the smooth muscle-to-collagen ratio within the corpus cavernosum was significantly improved in both of the SVF groups compared to vehicle-treated rats. The main limitation of the study is the lack of determination of the SVF components. CONCLUSIONS: Uncultured autologous SVF injected immediately or 4 wk after CN crush improved erectile function, promoted nerve regeneration, and prevented fibrosis of the corpus cavernosum following CN injury.
Assuntos
Tecido Adiposo/transplante , Disfunção Erétil/cirurgia , Músculo Liso/transplante , Pênis/lesões , Pênis/cirurgia , Células Estromais/transplante , Tecido Adiposo/citologia , Animais , Modelos Animais de Doenças , Masculino , Músculo Liso/citologia , Compressão Nervosa , Regeneração Nervosa , Óxido Nítrico Sintase Tipo I/biossíntese , Ereção Peniana/fisiologia , Pênis/citologia , Pênis/enzimologia , Pênis/inervação , Nervo Pudendo/citologia , Nervo Pudendo/metabolismo , Ratos , Ratos Sprague-Dawley , Recuperação de Função Fisiológica , Células Estromais/citologiaRESUMO
N-(3-hydroxy-1, 3, 5 (10) estratrien-17beta-yl)-3-hydroxypropylamine (17ß aminoestrogen, prolame) is a steroidal compound with weak estrogen-related trophic-proliferative effects in uterus. Contrasting with 17ß-estradiol (E2) pro-coagulant effects, this compound has high anticoagulant and antiplatelet effects. It has been extensively demonstrated that E2 plays important roles in brain function. However, prolame's influence on central nervous system has not been documented. In this study, we evaluated the effects of prolame replacement in young ovariectomized rats on spatial learning and memory and anxiety, correlating pyramidal cell dendritic spine density changes and neuronal nitric oxide synthase (nNOS) expression in the hippocampus. Ovariectomized young rats were treated with prolame for 4 weeks. Three other groups were used as physiological, pathological, and pharmacological references as follow: gonadally intact cycling females, ovariectomized, and ovariectomized with 17ß-estradiol treatment respectively, for the same time period. Experiment 1 investigated the behavioral effects of prolame on anxiety and spatial learning using elevated plus maze (EPM) and Morris water maze (MWM) paradigms respectively. Experiment 2 studied the dendritic spine density and neuronal nitric oxide synthase expression in the hippocampus of the 4 experimental groups. Similar to estradiol, prolame reversed the anxiogenic effects of ovariectomy, evaluated by EPM, and enhanced MWM performance to the level of gonadally intact subjects. Hippocampi from prolame-treated rats exhibited enhanced nNOS immunoreactivity and its relocation in dendritic compartments, as well as recovery of dendritic spine density loss in pyramidal neurons. Hence, prolame may provide an alternative option for ameliorating neurological symptoms caused by surgical menopause.
Assuntos
Ansiedade/tratamento farmacológico , Estrenos/uso terapêutico , Terapia de Reposição Hormonal/psicologia , Deficiências da Aprendizagem/tratamento farmacológico , Transtornos da Memória/tratamento farmacológico , Óxido Nítrico Sintase Tipo I/biossíntese , Ovariectomia/efeitos adversos , Animais , Espinhas Dendríticas/ultraestrutura , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos/métodos , Avaliação Pré-Clínica de Medicamentos/psicologia , Estradiol/farmacologia , Estradiol/uso terapêutico , Estrenos/síntese química , Estrenos/farmacologia , Feminino , Hipocampo/citologia , Hipocampo/metabolismo , Terapia de Reposição Hormonal/métodos , Aprendizagem em Labirinto/efeitos dos fármacos , Ratos , Ratos WistarRESUMO
Neuronal nitric oxide synthase (nNOS) has been reported to be up-regulated in the hypothalamic supraoptic nucleus (SON) during dehydration which in turn could increase nitric oxide (NO) production and consequently affect arginine vasopressin (AVP) secretion. The anteroventral third ventricle (AV3V) region has strong afferent connections with the SON. Herein we describe our analysis of the effects of an AV3V lesion on AVP secretion, and c-fos and nNOS expression in the SON following dehydration. Male Wistar rats had their AV3V region electrolytically lesioned or were sham operated. After 21 days they were submitted to dehydration or left as controls (euhydrated). Two days later, one group was anaesthetized, perfused and the brains were processed for Fos protein and nNOS immunohistochemistry (IHC). Another group was decapitated, the blood collected for hematocrit, osmolality, serum sodium and AVP plasma level analysis. The brains were removed for measurement of neurohypophyseal AVP content, and the SON was punched out and processed for nNOS detection by western blotting. The AV3V lesion reduced AVP plasma levels and c-fos expression in the SON following dehydration (P<0.05). Western blotting revealed an up-regulation of nNOS in the SON of control animals following dehydration, whereas such up-regulation was not observed in AV3V-lesioned rats (P<0.05). We conclude that the AV3V region plays a role in regulating the expression of nNOS in the SON of rats submitted to dehydration, and thus may affect the local nitric oxide production and the secretion of vasopressin.
Assuntos
Hipotálamo/enzimologia , Óxido Nítrico Sintase Tipo I/biossíntese , Terceiro Ventrículo/lesões , Privação de Água/fisiologia , Animais , Arginina Vasopressina/sangue , Desidratação/metabolismo , Hematócrito , Imuno-Histoquímica , Masculino , Concentração Osmolar , Neuro-Hipófise/metabolismo , Proteínas Proto-Oncogênicas c-fos/metabolismo , Ratos , Ratos Wistar , Sódio/sangue , Núcleo Supraóptico/metabolismo , Terceiro Ventrículo/patologia , Vasopressinas/metabolismoRESUMO
Experiment carried out on laboratory animals (rats) were aimed at comparative evaluation of the effect of several neuroprotective drugs under the conditions of model brain ischemia-reperfusion. The experimental methods included staining of brain tissue sections by hematoxiline-eosine, Nissl staining, and expression of NOS1, NOS3, TRAIL by imunnohistological means. The intensity of damage in various parts of brain and the nature of apoptosis without neuroprotection and with popular neuroprotectors (cytoflavin, actovegin, mexidol) and a test drug at the stage ofpreclinical trial (AKF-90-7) were evaluated. Characteristic cytotoxic (coagulative pycnomorphic and colliquative necrosis of neurons) and vascular (hemostasia, erythropedesis) changes were revealed. The neuroprotective effectof drugs decreases in the following order: AKF-90-7 > cytoflavin > actovegin > mexidol.
Assuntos
Encéfalo/efeitos dos fármacos , Glicina/análogos & derivados , Hemostasia/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Fármacos Neuroprotetores/uso terapêutico , Picolinas/uso terapêutico , Traumatismo por Reperfusão/tratamento farmacológico , Animais , Encéfalo/metabolismo , Encéfalo/patologia , Combinação de Medicamentos , Avaliação Pré-Clínica de Medicamentos , Amarelo de Eosina-(YS)/análise , Mononucleotídeo de Flavina/administração & dosagem , Mononucleotídeo de Flavina/uso terapêutico , Glicina/administração & dosagem , Glicina/uso terapêutico , Hematoxilina/análise , Heme/administração & dosagem , Heme/análogos & derivados , Heme/uso terapêutico , Imuno-Histoquímica , Inosina Difosfato/administração & dosagem , Inosina Difosfato/uso terapêutico , Masculino , Necrose/prevenção & controle , Neurônios/metabolismo , Neurônios/patologia , Fármacos Neuroprotetores/administração & dosagem , Niacinamida/administração & dosagem , Niacinamida/uso terapêutico , Óxido Nítrico Sintase Tipo I/análise , Óxido Nítrico Sintase Tipo I/biossíntese , Óxido Nítrico Sintase Tipo III/análise , Óxido Nítrico Sintase Tipo III/biossíntese , Picolinas/administração & dosagem , Ratos , Ratos Endogâmicos , Traumatismo por Reperfusão/sangue , Succinatos/administração & dosagem , Succinatos/uso terapêutico , Ligante Indutor de Apoptose Relacionado a TNF/análise , Ligante Indutor de Apoptose Relacionado a TNF/biossínteseRESUMO
Nitric oxide (NO)-containing neurons are widely distributed within the central nervous system, including regions involved in the control of reproduction and sexual behavior. Nitrergic neurons may co-localize with gonadal hormone receptors and gonadal hormones may influence neuronal NO synthase expression in adulthood as well as during development. In rodents, the female, in physiological conditions, is exposed to short-term changes of gonadal hormones levels (estrous cycle). Our studies, performed in mouse hypothalamic and limbic systems, reveal that the expression of neuronal NO synthase may vary according to the rapid variations of hormonal levels that take place during the estrous cycle. This is in accordance with the hypothesis that gonadal hormone activation of NO-cGMP pathway is important for mating behavior. NO-producing system appears particularly sensitive to alterations of endocrine balance during development, as demonstrated by our experiments utilizing perinatal exposure to bisphenol A, an endocrine disrupting chemical. In fact, significant effects were detected in adulthood in the medial preoptic nucleus and in the ventromedial subdivision of the bed nucleus of the stria terminalis. Therefore, alteration of the neuronal NO synthase expression may be one of the causes of the important behavioral alterations observed in bisphenol-exposed animals.
Assuntos
Comportamento Animal/fisiologia , Disruptores Endócrinos/toxicidade , Ciclo Estral/fisiologia , Hipotálamo/metabolismo , Neurônios Nitrérgicos/metabolismo , Óxido Nítrico/biossíntese , Animais , Comportamento Animal/efeitos dos fármacos , Compostos Benzidrílicos , Disruptores Endócrinos/metabolismo , Estrogênios não Esteroides/metabolismo , Estrogênios não Esteroides/toxicidade , Ciclo Estral/efeitos dos fármacos , Feminino , Hipotálamo/efeitos dos fármacos , Hipotálamo/enzimologia , Masculino , Camundongos , Neurônios Nitrérgicos/efeitos dos fármacos , Neurônios Nitrérgicos/enzimologia , Óxido Nítrico Sintase Tipo I/biossíntese , Fenóis/metabolismo , Fenóis/toxicidadeRESUMO
We examined the effect of high-fat (HF) feeding on blood pressure (BP) regulation, including hypothalamic redox signaling, as well as the changes in diurnal patterns and responses to restraint stress. Furthermore, we investigated whether HF feeding affects catecholamine and neuropeptide Y (NPY) biosynthesis in the adrenal medulla. Male obesity-prone Sprague-Dawley rats were fed with standard rat chow or 60% HF diet for 6 months. BP and heart rate (HR) were measured by telemetry, and circadian changes as well as responses to 20 min restraint stress were analyzed. Mean arterial BP was significantly elevated in HF rats both during daytime and nighttime compared with controls, whereas HR was elevated only during the day. BP and HR increased similarly in response to stress in both experimental groups; however, post-stress recovery of BP and HR were significantly delayed in HF animals. Protein levels of angiotensin II type 1 receptor (AT(1)) and NOX2, p67(phox) and p47(phox) subunits of NADPH oxidase, as well as NADPH oxidase activity increased significantly in the hypothalamus with HF feeding, whereas levels of antioxidant enzymes and nitric oxide synthases remained unchanged. In addition, HF diet also elevated the adrenomedullary protein levels of tyrosine hydroxylase and NPY. This study shows that feeding obesity-prone Sprague-Dawley rats with a HF diet results in elevated BP and HR and delayed cardiovascular post-stress recovery, and that these changes are paralleled by increases in the expression and activity of NADPH oxidase in the hypothalamus without a compensatory increase in the antioxidant enzyme levels, possibly leading to superoxide-mediated sympathoexcitation and hypertension.
Assuntos
Pressão Sanguínea/fisiologia , Gorduras na Dieta/farmacologia , Hipotálamo/efeitos dos fármacos , Hipotálamo/metabolismo , Transdução de Sinais/fisiologia , Medula Suprarrenal/efeitos dos fármacos , Angiotensina II/metabolismo , Angiotensina II/fisiologia , Animais , Northern Blotting , Western Blotting , Catecolaminas/metabolismo , Frequência Cardíaca/efeitos dos fármacos , Masculino , Atividade Motora/efeitos dos fármacos , NADPH Oxidases/metabolismo , Óxido Nítrico Sintase Tipo I/biossíntese , Óxido Nítrico Sintase Tipo II/biossíntese , Oxirredução , Ratos , Ratos Sprague-Dawley , Restrição Física , Estresse Psicológico/fisiopatologia , Telemetria , Tirosina 3-Mono-Oxigenase/biossínteseRESUMO
BACKGROUND: Nitric oxide plays an important role in the regulation of male and female sexual behavior in rodents, and the expression of the nitric oxide synthase (NOS) is influenced by testosterone in the male rat, and by estrogens in the female. We have here quantitatively investigated the distribution of nNOS immunoreactive (ir) neurons in the limbic hypothalamic region of intact female mice sacrificed during different phases of estrous cycle. RESULTS: Changes were observed in the medial preoptic area (MPA) (significantly higher number in estrus) and in the arcuate nucleus (Arc) (significantly higher number in proestrus). In the ventrolateral part of the ventromedial nucleus (VMHvl) and in the bed nucleus of the stria terminalis (BST) no significant changes have been observed. In addition, by comparing males and females, we observed a stable sex dimorphism (males have a higher number of nNOS-ir cells in comparison to almost all the different phases of the estrous cycle) in the VMHvl and in the BST (when considering only the less intensely stained elements). In the MPA and in the Arc sex differences were detected only comparing some phases of the cycle. CONCLUSION: These data demonstrate that, in mice, the expression of nNOS in some hypothalamic regions involved in the control of reproduction and characterized by a large number of estrogen receptors is under the control of gonadal hormones and may vary according to the rapid variations of hormonal levels that take place during the estrous cycle.
Assuntos
Ciclo Estral/metabolismo , Hipotálamo/enzimologia , Sistema Límbico/enzimologia , Óxido Nítrico Sintase Tipo I/biossíntese , Animais , Feminino , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos DBA , Neurônios/enzimologia , Caracteres SexuaisRESUMO
Striatal administration of 1-methyl-4-phenylpyridinium (MPP(+)), the active metabolite of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), causes nigrostriatal dopaminergic pathway damage similar to that observed in Parkinson's disease. Copper acts as a prosthetic group of several antioxidant enzymes and recent data show that copper attenuated MPP(+)-evoked neurotoxicity. We evaluated the effect of copper (as a supplement) upon proteins nitration (60 kDa) and tyrosine hydroxylase (TH) inactivation induced by MPP(+) (10 microg/8 microL) injection into the rat striatum. Copper pretreatment (10 micromol/kg i.p.) prevented both MPP(+)-induced proteins nitration and TH inactivation. Copper treatment also prevented the dopamine-depleting effect of MPP(+) injection. Those results were accompanied by a significant reduction of enzymatic activity of the constitutive nitric oxide synthase (cNOS), whereas, the protein levels of the three isoforms of NOS remained unchanged. Results indicate that the effect of copper against MPP(+)-induced proteins nitration and TH inactivation in the striatum of rat may be mediated by a reduction of cNOS activity.
Assuntos
Sulfato de Cobre/farmacologia , Intoxicação por MPTP/prevenção & controle , Neostriado/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Nitratos/metabolismo , Tirosina 3-Mono-Oxigenase/antagonistas & inibidores , 1-Metil-4-Fenil-1,2,3,6-Tetra-Hidropiridina , Animais , Western Blotting , Dopamina/metabolismo , Dopaminérgicos/toxicidade , Eletroforese em Gel de Poliacrilamida , Masculino , Neostriado/efeitos dos fármacos , Óxido Nítrico Sintase Tipo I/biossíntese , Doença de Parkinson Secundária/prevenção & controle , Ratos , Ratos WistarRESUMO
NO (nitric oxide) is a highly reactive free radical gas thought to play a major role in the invertebrate immune response by harming pathogens and limiting their growth. Here we report on studies of nitric oxide synthase (NOS) genes in the crustacean Daphnia, one of the few non-insect arthropod models used to study host-pathogen interactions. While the NOS gene is found as a single copy in other invertebrates, we found two copies (NOS1 and NOS2), which a phylogenetic reconstruction showed to be the result of an ancient duplication event. Both genes bear features commonly found in invertebrate NOS, however, the two genes differ in their rate of evolution, intraspecific polymorphism and expression level. We tested whether the more rapid evolution of NOS2 could be due to positive selection, but found the rate of amino-acid substitutions between Daphnia species to be compatible with a neutral model. To associate NOS or NO activity with infection, we performed infection experiments with Daphnia magna and one of its natural pathogens (the bacterium Pasteuria ramosa). In one set of experimental infections, we supplemented D. magna with L-arginine, the NOS substrate, or with L-NAME, a NOS antagonist, and found this to result in lower and higher infection levels, respectively, which is at least compatible with the notion that NO may aid defence against Pasteuria. A second experiment indicated that NOS transcription does not increase following exposure to Pasteuria. Thus, the function of NOS in Daphnia immunity remains uncertain, but the pattern of gene duplication and subsequent divergence suggests evolution via neo- or subfunctionalization.
Assuntos
Daphnia/genética , Daphnia/imunologia , Duplicação Gênica , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo I/genética , Sequência de Aminoácidos , Animais , Bactérias , Sequência de Bases , Daphnia/microbiologia , Evolução Molecular , Dados de Sequência Molecular , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo I/biossíntese , Óxido Nítrico Sintase Tipo I/classificação , Óxido Nítrico Sintase Tipo II/biossíntese , Óxido Nítrico Sintase Tipo II/classificação , Filogenia , Polimorfismo Genético , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Alinhamento de SequênciaRESUMO
The role of nitric oxide synthases (NOS) and the nitric oxide (NO) substrate l-arginine in renal ischaemia-reperfusion (I/R) has been studied extensively. However, the results reported are often controversial. In the present study, we examined the effects of the neuronal (n) NOS inhibitor 7-nitroindazole (7-NI) and L-arginine administration on renal I/R injury and the renal NO system in rats. Following 7 days pretreatment with 7-NI (50 mg/kg per day), L-arginine (2 g/kg per day) or vehicle (dimethylsulphoxide : sesame oil, 1 : 9), the left renal vascular pedicles were clamped for 50 min in male Sprague-Dawley rats and kidneys were removed 24 h after reperfusion (n = 7/group). Neither 7-NI nor L-arginine had any effect on parameters of renal function, the grade of tissue injury or the number of terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end-labelling (TUNEL)-positive tubular cells compared with vehicle-treated rats. 7-Nitroindazole decreased nNOS mRNA expression and inducible (i) NOS protein levels, but had no effect on endothelial NOS expression. L-arginine supplementation increased mRNA expression of all NOS isoforms, but only increased protein expression of iNOS. The results of the present study demonstrate that selective inhibition of nNOS has no effect on renal injury, indicating that nNOS does not play a central role in the pathophysiology of renal I/R. In addition, although L-arginine has no effect on renal I/R injury in the model used in the present study, its administration increases the mRNA expression of NOS isoforms.
Assuntos
Arginina/administração & dosagem , Rim/irrigação sanguínea , Rim/enzimologia , Óxido Nítrico Sintase Tipo I/antagonistas & inibidores , Óxido Nítrico/fisiologia , Traumatismo por Reperfusão/enzimologia , Animais , Arginina/fisiologia , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Indazóis/administração & dosagem , Indazóis/farmacologia , Isoenzimas/biossíntese , Isoenzimas/genética , Rim/efeitos dos fármacos , Rim/fisiopatologia , Masculino , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo I/biossíntese , Óxido Nítrico Sintase Tipo I/metabolismo , RNA Mensageiro/biossíntese , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Traumatismo por Reperfusão/tratamento farmacológico , Traumatismo por Reperfusão/fisiopatologia , Especificidade por Substrato/fisiologia , Regulação para Cima/fisiologiaRESUMO
OBJECTIVE: To detect the expression of HO-2 in the corpus cavernosum of rats with chronic renal failure (CRF) , and investigate the role of HO-2 in penile erection and its association with testosterone. METHODS: Fifteen 10-week-old SD rats underwent 5/6 kidney removal for the establishment of CRF models, and another 15 included as controls. Twelve weeks later, both the two groups of animals were subjected to electrostimulation of the cavernous nerve for the detection of intracavernosal pressure (ICP) and mean arterial pressure (MAP), and the protein contents of HO-2, nNOS and eNOS in the penile tissues were determined by Western blot and immunohistochemical analysis. RESULTS: The ICPmax/MAP after 3 V and 5 V stimulation of the cavernous nerve was (0.121 +/- 0.084) and (0.135 +/- 0.088), the serum testosterone level was (1.190 +/- 0.946) nmol/L, and the expression of HO-2 was (0.510 +/- 0.397) in the CRF group, all significantly lower than in the control rats, which were (0.263 +/- 0.147 and 0.244 +/- 0.089), (7.800 +/- 5.001) nmol/L (P<0.01) and (2.672 +/- 1.720, P<0.01), respectively. There was a correlation between the decrease of HO-2 expression and the reduction of serum testosterone (r = 0.902, P < 0.01). CONCLUSION: The lowered level of serum testosterone and decreased contents of HO-2, eNOS and nNOS may play a role in CRF-induced ED.
Assuntos
Disfunção Erétil/fisiopatologia , Heme Oxigenase (Desciclizante)/biossíntese , Falência Renal Crônica/fisiopatologia , Pênis/enzimologia , Animais , Western Blotting , Disfunção Erétil/complicações , Imuno-Histoquímica , Falência Renal Crônica/complicações , Masculino , Óxido Nítrico Sintase Tipo I/biossíntese , Óxido Nítrico Sintase Tipo III/biossíntese , Ratos , Ratos Sprague-DawleyRESUMO
OBJECTIVE: To determine the effects of Tongxinluo on cell viability and tissue factor (TF) in AngII induced vascular endothelial cells and to investigate its mechanism. METHODS: AngII(10(-6)mol/L) was added to human vascular endothelial cells (HUVECs) culture media alone or with various concentration of Tongxinluo drug containing plasma (5%,10%, and 20%) added 30 minutes before AngII. Cell viability was evaluated after 24-hour incubation with AngII in a dose manner. TF, AngII type 1 receptor (AT(1)) mRNA, NO synthase (NOS) and NO were observed after 24-hour incubation with AngII. In addition, NOS inhibitor nomega-nitro-larginine (L-NAME) was added 30 minutes before Tongxinluo and AngII. Cell viability, TF, AT(1)mRNA, the level of NOS and NO were evaluated after 24-hour incubation with Tongxinluo and AngII. RESULTS: Tongxinluo significantly improved AngII induced endothelial cell viability and the effect was the most obvious at 10%. Tongxinluo (10%) decreased the TF and AT(1) mRNA while increased the NOS and NO levels. L-NAME obviously inhibited the effects of Tongxinluo on cell viability, TF, AT(1) mRNA, and NOS and NO levels. CONCLUSION: Up-regulating NOS-NO signaling may be the mechanism of Tongxinluo on cell viability and TF in AngII induced vacular endothelial cells.
Assuntos
Angiotensina II/farmacologia , Medicamentos de Ervas Chinesas/farmacologia , Endotélio Vascular/efeitos dos fármacos , Tromboplastina/biossíntese , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Endotélio Vascular/citologia , Endotélio Vascular/metabolismo , Inibidores Enzimáticos , Ensaio de Imunoadsorção Enzimática , Humanos , NG-Nitroarginina Metil Éster/farmacologia , Óxido Nítrico Sintase Tipo I/antagonistas & inibidores , Óxido Nítrico Sintase Tipo I/biossíntese , Óxido Nítrico Sintase Tipo I/genética , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Receptor Tipo 1 de Angiotensina/biossíntese , Receptor Tipo 1 de Angiotensina/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Tromboplastina/genéticaRESUMO
Viscum album L. aqueous extract, on the Langendorff isolated and perfused heart model, decreases coronary vascular resistance, when compared to control group (36.00 +/- 2.00 vs. 15.80 +/- 1.96 dyn s cm-5). Our data support the fact that this mechanism involves NOS-2 and NOS-3 overexpression (4.65 and 7.89 times over control, respectively), which is correlated with increases in NO (6.24 +/- 2.49 vs. 147.95 +/- 2.79 pmol) and cGMP production (43.94 +/- 2.00 vs. 74.81 +/- 1.96 pmol mg-1 of tissue), compared to control values. Such an effect is antagonized by gadolinium(III) chloride, L-NAME and ODQ. Therefore, coronary vasodilator effect elicited by V. album L. aqueous extract is mediated by the NO/sGC pathway.
Assuntos
Miocárdio/enzimologia , Óxido Nítrico Sintase Tipo III/biossíntese , Óxido Nítrico Sintase Tipo I/biossíntese , Extratos Vegetais/farmacologia , Viscum album/química , Animais , Vasos Coronários/efeitos dos fármacos , Indução Enzimática/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Guanilato Ciclase/metabolismo , Cobaias , Técnicas In Vitro , Masculino , Contração Miocárdica/efeitos dos fármacos , Miocárdio/metabolismo , Óxido Nítrico/biossíntese , Óxido Nítrico Sintase Tipo I/antagonistas & inibidores , Óxido Nítrico Sintase Tipo III/antagonistas & inibidores , Folhas de Planta/metabolismo , Resistência Vascular/efeitos dos fármacosRESUMO
This study was to investigate the antinociceptive effects of moxibustion in a complete Freund's adjuvant (CFA)-induced arthritic rat model, and the effects of moxibustion on immunohistochemical changes at the spinal cord level. Moxibustion was applied to the ipsilateral (right) Zusanli (ST36) acupoint to the lesion side for 9 days to CFA-induced arthritic rats. The stepping force was measured as a behavioral test, c-Fos immunohistochemistry, NO production and nNOS Western blots were examined to evaluate antinociceptive effects. Moxibustion at ST36 significantly improved the stepping force in the affected hind limb in CFA-induced arthritis. Moreover, moxibustion at ST36 suppressed the production of NO and the protein expression of c-Fos and nNOS induced by arthritis. These results suggest that moxibustion at ST36 has a potent antinociceptive effect in an arthritic rat model, and modulates neuronal excitability and endogenous NO production by suppressing c-Fos and nNOS protein expression.
Assuntos
Pontos de Acupuntura , Artrite Experimental/terapia , Moxibustão , Adjuvantes Imunológicos/farmacologia , Animais , Artrite Experimental/metabolismo , Modelos Animais de Doenças , Adjuvante de Freund/farmacologia , Membro Posterior , Masculino , Atividade Motora , Óxido Nítrico/biossíntese , Óxido Nítrico Sintase Tipo I/biossíntese , Proteínas Proto-Oncogênicas c-fos/biossíntese , Ratos , Ratos Sprague-DawleyRESUMO
Nitric oxide synthase (NOS) and interleukin-6 (IL-6) are constitutively expressed in hypothalamic cells. However, phenotypic and functional aspects of these cells remain unknown. We have studied the expression pattern of these two molecules in hypothalamic cells expressing corticotropin-releasing factor (CRF) and arginin-vasopressin (AVP), two major regulatory peptides in the hypothalamus-pituitary system, using immunofluorescence, intracerebroventricular injection of colchicine, and the study in parallel of the labeling pattern of axons in the median eminence. Within AVP cells, we distinguished two different populations: large, intensely stained AVP cells coexpressing IL-6; and large, intensely stained AVP cells coexpressing IL-6 and NOS. Within the CRF cells, we distinguished three different populations: large, intensely stained CRF cells immunonegative for AVP, NOS, and IL-6; large cells weakly stained for CRF and AVP, immunopositive for NOS and immunonegative for IL-6; and small cells intensely stained for CRF and AVP and immunonegative for IL-6 and NOS. In addition, we also found AVP cells containing IL-6 in the suprachiasmatic nucleus. These results suggest that neuronal NOS and IL-6 may be involved in different modulatory processes in hypophysiotropic and non-hypophysiotropic cells.
Assuntos
Arginina Vasopressina/metabolismo , Hormônio Liberador da Corticotropina/metabolismo , Hipotálamo/metabolismo , Interleucina-6/biossíntese , Óxido Nítrico Sintase Tipo I/biossíntese , Animais , Colchicina/administração & dosagem , Colchicina/farmacologia , Hipotálamo/anatomia & histologia , Imuno-Histoquímica , Injeções Intraventriculares , Masculino , Eminência Mediana/metabolismo , Núcleo Hipotalâmico Paraventricular/metabolismo , Ratos , Ratos Sprague-Dawley , Núcleo Supraquiasmático/metabolismo , Núcleo Supraóptico/metabolismoRESUMO
OBJECTIVE: To investigate the effect of Niupo Zhibao Pellet (NPZBP) on the expression of neuronal nitric oxide synthase (nNOS) in the brain of endotoxin-induced shock rats. METHODS: SD rats were randomly divided into normal control group, endotoxin-induced shock model group and NPZBP-treated group. Lipopolysaccharide (LPS) (1.5 mg/kg i.v.) and tsD-galactosamine (D-GalN) (100 mg/kg i.p.) were administered to the rats in endotoxin-induced shock model group, as well as to the rats in NPZBP-treated group after seven-day treatment, to induce the shock. The expression of nNOS in the brain of the rats in each of the 3 groups was measured by immunohistochemical methods. RESULTS: In the 3 groups, nNOS immuno-positive cells distributed widely in layer II, III, IV of the cerebral cortex, the molecular layer of hippocampus, the polymorphic layer of the dentate gyrus, the reticular formation of brain stem, and the molecular, granular and Purkinje cell layer of the cerebellar cortex. The number of immuno-positive cells in the NPZBP-treated group was slightly higher than that of the normal control group, and significantly lower than that of the model group (P<0.05) in many regions of the brain, including cerebral cortex, hippocampus, brain stem and cerebellar cortex. CONCLUSION: NPZBP can inhibit the over-expression of nNOS in wide area of the brain in endotoxin-induced shock rats.