RESUMO
Most studies on the effect of tibolone on the uterus have focused on the endometrium dismissing the importance of the myometrium. The aim of the present study was to investigate some estrogen-like actions of tibolone in the uterus assessed by: 1) the expression of estrogen, progesterone, and serotonin receptors, and 2) the myometrial contraction induced by serotonin. Estradiol (250 µg), progesterone (50 mg), or testosterone (25 mg) pellets were implanted to ovariectomized rats. Tibolone (0.5 mg/day) was orally administered. An implanted pellet containing vehicle or an equivalent volume of water p.o., were used as controls. Sixty days after beginning the treatments, rats were killed and uterus removed. One horn was processed to evaluate estrogen-alpha, progesterone A and B, and serotonin-2A receptors expression, and the other one was used for studying contraction to serotonin and 60 mM potassium solution. The present data showed that tibolone-induced expression of estrogen, progesterone, and serotonin receptors, but did not induce uterine contractile response to either serotonin or potassium solution. These findings suggest that, in the uterus, tibolone may exert molecular estrogenic actions such as the induction of receptor expression, but not a physiological response as the estrogen-dependent contraction to serotonin.
Assuntos
Receptor alfa de Estrogênio/genética , Expressão Gênica/efeitos dos fármacos , Norpregnenos/farmacologia , Receptores de Progesterona/genética , Receptores de Serotonina/genética , Contração Uterina/efeitos dos fármacos , Útero/efeitos dos fármacos , Animais , Receptor alfa de Estrogênio/metabolismo , Estrogênios/farmacologia , Feminino , Ratos , Ratos Sprague-Dawley , Receptores de Progesterona/metabolismo , Receptores de Serotonina/metabolismo , Útero/fisiologiaRESUMO
BACKGROUND: A hallmark of airway remodelling in asthma is subepithelial fibrosis, but its relation with airway dysfunction is still controversial. OBJECTIVE: To describe airway functional abnormalities and subepithelial remodelling induced by repetitive antigenic challenges. METHODS: Nine inhaled antigenic challenges were applied every 10 days to guinea-pigs sensitized to ovalbumin (OVA). Antigen-induced airway hyperresponsiveness (AI-AHR) to histamine and its immunohistopathological relationship was evaluated at the first, third and ninth OVA challenges. RESULTS: From the first challenge on, OVA induced acute transient bronchoobstruction followed by development of AI-AHR. A progressive rise in baseline Penh (a bronchoobstruction index) and granulocyte airway infiltration was also observed. After the ninth OVA challenge, the amount of extracellular matrix in the subepithelial region (SER) of bronchi and bronchioles was increased. Immunohistochemistry analysis showed that this SER fibrosis was associated to beta1-integrin subunit overexpression, even in acellular areas. Immunoelectronmicroscopy corroborated the location of beta1-integrin in extracellular matrix, essentially in types l and II collagen fibres. Presence of alpha1- and alpha2-integrin subunits in these areas was also corroborated. AI-AHR was correlated with degree of SER increment, cell infiltration, and beta1-integrin expression. CONCLUSION: Our data suggested that beta1-integrin shedding produced by repetitive allergen challenges in guinea-pigs was associated with collagen deposition in SER of bronchi and bronchioles, along with inflammatory cells infiltration and AI-AHR development.
Assuntos
Asma/imunologia , Brônquios/imunologia , Integrina beta1/metabolismo , Administração por Inalação , Animais , Asma/patologia , Brônquios/ultraestrutura , Líquido da Lavagem Broncoalveolar/imunologia , Doença Crônica , Colágeno/imunologia , Colágeno/metabolismo , Modelos Animais de Doenças , Matriz Extracelular/imunologia , Matriz Extracelular/metabolismo , Cobaias , Integrina beta1/imunologia , Masculino , Microscopia Imunoeletrônica , Ovalbumina/imunologiaRESUMO
Di(2-ethylhexyl) phthalate (DEHP) is a widely used plasticizer that is metabolized to mono(2-ethylhexyl) phthalate (MEHP). Inhalation is an important exposure route for both phthalates, and their effects on lungs include inflammation, alteration of postnatal maturation (alveolarization), enlarged airspaces and cell differentiation changes, suggesting that alveolar epithelial cells-2 (AEC) are targets of phthalates. This study evaluated the cell progression, epithelial and mesenchymal markers, including surfactant secretion in A549 cells (AEC) that were exposed to DEHP (1-100⯵M) or MEHP (1-50⯵M) for 24-72â¯h. The results showed an increased cell proliferation at all concentrations of each phthalate at 24 and 48â¯h. Cell migration showed a concentration-dependent increase at 24 and 48â¯h of exposure to either phthalate and enlarged structures were seen. Decreased levels of both surfactants (SP-B/SP-C) were observed after the exposure to either phthalate at 48â¯h, and of SP-C positive cells exposed to MEHP, suggesting a loss of the epithelial phenotype. While a decrease in the epithelial marker E-cadherin and an increase in the mesenchymal marker fibronectin were observed following exposure to either phthalate. Our results showed that DEHP and MEHP altered the structure and migration of A549 cells and promoted the loss of the epithelial phenotype.
Assuntos
Células Epiteliais Alveolares/efeitos dos fármacos , Desdiferenciação Celular/efeitos dos fármacos , Dietilexilftalato/análogos & derivados , Dietilexilftalato/toxicidade , Plastificantes/toxicidade , Proteína B Associada a Surfactante Pulmonar/antagonistas & inibidores , Proteína C Associada a Surfactante Pulmonar/antagonistas & inibidores , Células A549 , Células Epiteliais Alveolares/citologia , Células Epiteliais Alveolares/metabolismo , Antígenos CD , Biomarcadores/metabolismo , Caderinas/antagonistas & inibidores , Caderinas/metabolismo , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Forma Celular/efeitos dos fármacos , Tamanho Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Fibronectinas/agonistas , Fibronectinas/metabolismo , Humanos , Cinética , Proteína B Associada a Surfactante Pulmonar/metabolismo , Proteína C Associada a Surfactante Pulmonar/metabolismoRESUMO
We studied the effect of transections at the superior cerebellar peduncle during the evolution of amygdaloid kindling. Dentato- and interposito-thalamic pathways, including the ascending fastigial fibers, were transected in 10 rats at the contralateral side of the stimulated amygdala, and in other 8 at the ipsilateral side. A group of 18 rats was used as control. Contralateral lesion significantly slowed amygdala kindling, while ipsilateral lesion decreased kindled seizure duration. Furthermore, when kindled seizures were reached by 6 control rats, transection of the ipsilateral superior cerebellar peduncle led to reduction of subsequent seizures. These specific effects produced by transection of the superior cerebellar peduncle suggest that the cerebellum could exert a tonic effect over the participating circuitry used by the kindling process.
Assuntos
Tonsila do Cerebelo/fisiologia , Cerebelo/fisiologia , Excitação Neurológica , Convulsões/etiologia , Animais , Comportamento Animal/fisiologia , Eletrofisiologia , Masculino , Ratos , Ratos Endogâmicos , Convulsões/fisiopatologia , Fatores de TempoRESUMO
The effects of the exposure to ozone in the central nervous system are unknown, as it is doubtful if ozone enters beyond the respiratory tract. However, ozone exposure impairs human performance and induces subjective complaints such as fatigue, lethargy and headache. We studied electrographic aspects of sleep-wake organization in cats, and found that paradoxical sleep was promptly reduced during ozone exposure, followed by a dose-related increase of slow-wave sleep. These findings suggest that high concentrations of ozone or its reaction products induce striking changes in sleep patterns.
Assuntos
Ozônio/farmacologia , Sono/efeitos dos fármacos , Animais , Gatos , Relação Dose-Resposta a Droga , Masculino , Vigília/efeitos dos fármacosRESUMO
We evaluated the role of protein kinase C (PKC) in the sustained bronchial contraction (SBC) induced by carbachol (Cch) or histamine in a Ca2+-free medium and the possibility that each agonist uses a different Ca2+ store for this response. We studied third-order bronchi and airway smooth muscle (ASM) from first-order bronchi dissected free of cartilage and epithelium. Bronchial and ASM responsiveness to Cch or histamine were evaluated in Krebs solution (2.5 mM Ca2+) and in Ca2+-free medium. Cch and histamine induced an SBC in bronchial tissues in Ca2+-free medium. In ASM each agonist produced a transient contraction, but the response to histamine was much smaller. Cch induced a concentration-dependent accumulation of inositol phosphates (IPs) in both bronchi and ASM; however, histamine did not induce significant accumulation of IPs. Repeated exposure to histamine in bronchial rings abolished contractile responses in Ca2+-free media, but Cch added afterwards still produced a sustained contraction. This response was blocked when bronchial tissues were preincubated with 10 microM cyclopiazonic acid (CPA). Brief incubation of these preparations with a high EGTA concentration (1 mM) abolished the histamine-induced SBC. The SBC induced by Cch or histamine in Ca2+-free medium was not affected by the preincubation of the tissues with calphostin C, chelerythrine or staurosporine. We concluded that Cch mobilizes Ca2+ from two different sources during the SBC in Ca2+-free medium: from a CPA-sensitive one from sarcoplasmic reticulum (SR) and from a putative extracellular membrane Ca2+ pool sensitive to 1 mM EGTA, and neither process involved PKC activation. Histamine appeared to utilize the extracellular membrane pool only.
Assuntos
Brônquios/efeitos dos fármacos , Broncoconstrição/efeitos dos fármacos , Cálcio/farmacologia , Animais , Brônquios/metabolismo , Brônquios/fisiologia , Cálcio/metabolismo , Carbacol/farmacologia , Cardiotônicos/farmacologia , Cães , Inibidores Enzimáticos/farmacologia , Feminino , Histamina/farmacologia , Técnicas In Vitro , Fosfatos de Inositol/metabolismo , Masculino , Contração Muscular/efeitos dos fármacos , Músculo Liso/efeitos dos fármacos , Músculo Liso/metabolismo , Músculo Liso/fisiologia , Naftalenos/farmacologia , Proteína Quinase C/antagonistas & inibidoresRESUMO
Bordetella pertussis (BP) has been used as adjuvant for experimental animal immunization, but its effects on airway responsiveness are uncertain. Three groups of guinea pigs were used: animals with a single exposure to inhaled BP vaccine (strain 134, total dose 1.24 x 10(12) germs), animals submitted to a sensitization procedure through inhalation of ovalbumin plus BP, and healthy control animals. Four weeks after inhalation of BP or after the beginning of sensitization, dose- or concentration-response curves to histamine were constructed in vivo and in vitro (tracheal and parenchymal preparations). We found that BP alone produced lower responses to histamine than control guinea pigs in vivo (insufflation pressure, p = 0.0003) and in tracheal tissues (p = 0.04), but not in parenchymal preparations. Sensitization did not modify the responsiveness compared with their respective controls. These results suggest that some BP component(s), probably pertussis toxin, causes a long lasting airway hyporesponsiveness in guinea pigs.
Assuntos
Bordetella pertussis/imunologia , Hiper-Reatividade Brônquica/fisiopatologia , Vacina contra Coqueluche/administração & dosagem , Vacina contra Coqueluche/farmacologia , Adjuvantes Imunológicos/farmacologia , Administração por Inalação , Animais , Hiper-Reatividade Brônquica/etiologia , Relação Dose-Resposta a Droga , Cobaias , Histamina/farmacologia , Imunização , MasculinoRESUMO
Prejunctional and postjunctional effects of several ozone (O3) concentrations, including those found in highly polluted cities, were evaluated in guinea pig airways. Animals bred in O3-free conditions were exposed to air or O3 (0.3, 0.6 or 1.2 ppm) during 4 h, and studied 16-18 h later. Tracheal and bronchial rings were studied in organ baths. Electrical field stimulation (EFS) (100 V, 2 ms, 10 s) was given at increasing frequencies (0.25-16 Hz). Some tissues received atropine (2 microM) and/or propranolol (10 microM). Concentration-response curves to carbachol, isoproterenol, nitroprusside, and substance P were constructed. In tracheas, almost all O3 concentrations decreased the relaxation at low EFS frequencies, but had no effect on the propranolol-resistant (i-NANC) relaxation, suggesting that only adrenergic relaxation was affected. This was a prejunctional effect, since O3 did not modify the responses to isoproterenol. Relaxation induced by a nitric oxide (NO) donor, nitroprusside, was not affected by O3, which agrees with the lack of O3-effect on i-NANC system. O3 did not modify the EFS-induced e-NANC contraction in atropine-treated bronchi, nor the contraction caused by exogenous substance P. By contrast, in bronchi without atropine, 1.2 ppm O3 increased the e-NANC contraction induced by the highest EFS (16 Hz). O3 increased the maximum responses to carbachol in tracheas (1.2 ppm) and bronchi (0.6 and 1.2 ppm). In conclusion, we found that: a) O3 decreased adrenergic relaxation in guinea pig tracheas at low EFS frequencies through a prejunctional alteration; b) O3 did not modify the i-NANC relaxation in tracheas, at least the NO-mediated; c) O3 added a cholinergic component to the bronchial slow-phase (e-NANC) contraction evoked by EFS; and d) O3 enhanced the cholinergic responses in trachea and bronchi by a postjunctional mechanism.
Assuntos
Músculo Liso/efeitos dos fármacos , Junção Neuromuscular/efeitos dos fármacos , Oxidantes Fotoquímicos/toxicidade , Ozônio/toxicidade , Fibras Adrenérgicas , Agonistas Adrenérgicos beta/farmacologia , Antagonistas Adrenérgicos beta/farmacologia , Animais , Atropina/farmacologia , Brônquios , Carbacol/farmacologia , Fibras Colinérgicas , Relação Dose-Resposta a Droga , Estimulação Elétrica , Cobaias , Técnicas In Vitro , Isoproterenol/farmacologia , Masculino , Agonistas Muscarínicos/farmacologia , Antagonistas Muscarínicos/farmacologia , Contração Muscular/efeitos dos fármacos , Relaxamento Muscular/efeitos dos fármacos , Propranolol/farmacologia , TraqueiaRESUMO
BACKGROUND: In airway smooth muscle (ASM), Ca2+ influx in response to the Ca2+ depletion of the sarcoplasmic reticulum (SR) seems to play a role in the regulation of intracellular free Ca2+ concentrations ([Ca2+](i)). This study evaluates some possible Ca2+ entry pathways activated during SR-Ca2+ depletion induced by 10 mM caffeine. METHODS: Enzymatically dispersed bovine ASM cells were loaded with Fura-2/AM to permit measurement of [Ca2+](i) changes in single cells. RESULTS: Caffeine (10 mM) induced a transient increase in [[Ca2+](i) that depleted SR-Ca(2)+ content. After caffeine washout, a decrease in basal [Ca2+](i) (undershoot) was invariably observed, followed by a slow recovery. This phenomenon was inhibited by cyclopiazonic acid (5 microM). External Ca(2)+ removal in depolarized and nondepolarized cells induced a decrease in basal [Ca2+](i) that continued until depletion of the SR-Ca2+ content. The decrease in [Ca2+](i) induced by Ca2+-free physiological saline solution (PSS) was accelerated in caffeine-stimulated cells. Recovery from undershoot was not observed in Ca2+-free PSS. Depolarization with KCl and addition of D600 (30 microM) did not modify recovery. Similar results were obtained when the Na(+)/Ca2+ exchanger was blocked by substituting NaCl with KCl in normal PSS (Na(+)-free PSS) or by adding benzamil amiloride (25 microM). CONCLUSIONS: SR-Ca2+ content plays an important role in the Ca2+ leak induced by Ca2+-free medium, and does not depend on membrane potential. Additionally, recovery from undershoot after caffeine depends on extracellular Ca2+, and neither voltage-dependent Ca2+ channels nor the Na(+)/Ca2+ exchanger are involved.
Assuntos
Cafeína/farmacologia , Canais de Cálcio/efeitos dos fármacos , Sinalização do Cálcio/efeitos dos fármacos , Cálcio/metabolismo , Músculo Liso/efeitos dos fármacos , Retículo Sarcoplasmático/efeitos dos fármacos , Traqueia/citologia , Amilorida/análogos & derivados , Amilorida/farmacologia , Animais , Canais de Cálcio/fisiologia , Carbacol/farmacologia , Bovinos , Células Cultivadas , Galopamil/farmacologia , Indóis/farmacologia , Transporte de Íons/efeitos dos fármacos , Ionomicina/farmacologia , Ionóforos/farmacologia , Músculo Liso/citologia , Cloreto de Potássio/farmacologia , Retículo Sarcoplasmático/metabolismo , Trocador de Sódio e Cálcio/efeitos dos fármacos , Trocador de Sódio e Cálcio/fisiologiaRESUMO
Ozone (O(3)) causes airway hyperresponsiveness, but few studies have evaluated this effect at urban concentrations. In this work dose-response curves to intravenous acetylcholine, histamine or substance P were performed in guinea pigs with or without previous exposure to O(3) (0.15, 0.3, 0.6 or 1.2 ppm for 4 h, 16-18 h before the studies). We found airway hyperresponsiveness to histamine, but not to acetylcholine, only after 1.2 ppm O(3). By contrast, airway hyperresponsiveness to substance P was developed at O(3) levels encountered in highly-polluted cities (0.3 ppm). These results suggest that excitatory non-adrenergic non-cholinergic responses could be affected by air pollution, and that substance P is a useful pharmacological tool for evaluating the airway hyperresponsiveness induced by low O(3) concentrations.