RESUMO
TNFa has been associated with both, tumor survival and apoptosis. This cytokine is also involved in promoting cell migration during wound healing and tumorigenesis. SW756 is a HPV18-positive cervical carcinoma cell line, which has been used to study different mechanisms of cervical cancer progression. An in vitro assay of scratch wound healing onto monolayers of SW756 cells was used to assess the effect of TNFa on cell migration into a wound space. It was found that SW756 cells have the ability to migrate, but not proliferate in response to scratch wounding in a serum-free medium supplemented with TNFa. RT-PCR analysis showed that SW756 cells express TNFa mRNA when incubated in medium with and without serum. Wound closure and migration rate of SW756 cells were significantly increased in the presence of serum-free media supplemented with TNFa (10 ng/mL) as compared to serum-free media, and media supplemented with either anti-TNFa antibody or both TNFa and anti-TNFa antibody (p<0.05). The results showed a stimulatory effect of TNFa on the migration of SW756 cervical carcinoma cells, suggesting a novel and important role for TNFa in cervical cancer progression.
Assuntos
Feminino , Humanos , Carcinoma/microbiologia , Movimento Celular/efeitos dos fármacos , /genética , Fator de Necrose Tumoral alfa/farmacologia , Neoplasias do Colo do Útero/microbiologia , Linhagem Celular Tumoral , Meios de Cultura Livres de Soro , Carcinoma/genética , Carcinoma/patologia , Proliferação de Células/efeitos dos fármacos , /isolamento & purificação , Processamento de Imagem Assistida por Computador , Cinética , Microscopia de Vídeo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , RNA Mensageiro/metabolismo , Proteínas Recombinantes/efeitos dos fármacos , Neoplasias do Colo do Útero/genética , Cicatrização/efeitos dos fármacosRESUMO
TNFalpha has been associated with both, tumor survival and apoptosis. This cytokine is also involved in promoting cell migration during wound healing and tumorigenesis. SW756 is a HPV18-positive cervical carcinoma cell line, which has been used to study different mechanisms of cervical cancer progression. An in vitro assay of scratch wound healing onto monolayers of SW756 cells was used to assess the effect of TNFalpha on cell migration into a wound space. It was found that SW756 cells have the ability to migrate, but not proliferate in response to scratch wounding in a serum-free medium supplemented with TNFalpha. RT-PCR analysis showed that SW756 cells express TNFalpha mRNA when incubated in medium with and without serum. Wound closure and migration rate of SW756 cells were significantly increased in the presence of serum-free media supplemented with TNFalpha (10 ng/mL) as compared to serum-free media, and media supplemented with either anti-TNFalpha antibody or both TNFalpha and anti-TNFalpha antibody (p < 0.05). The results showed a stimulatory effect of TNFalpha on the migration of SW756 cervical carcinoma cells, suggesting a novel and important role for TNFalpha in cervical cancer progression.
Assuntos
Carcinoma/microbiologia , Movimento Celular/efeitos dos fármacos , Papillomavirus Humano 18/genética , Fator de Necrose Tumoral alfa/farmacologia , Neoplasias do Colo do Útero/microbiologia , Carcinoma/genética , Carcinoma/patologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Meios de Cultura Livres de Soro , Feminino , Papillomavirus Humano 18/isolamento & purificação , Humanos , Processamento de Imagem Assistida por Computador , Cinética , Microscopia de Vídeo , RNA Mensageiro/metabolismo , Proteínas Recombinantes/efeitos dos fármacos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Neoplasias do Colo do Útero/genética , Cicatrização/efeitos dos fármacosRESUMO
TNFalpha has been associated with both, tumor survival and apoptosis. This cytokine is also involved in promoting cell migration during wound healing and tumorigenesis. SW756 is a HPV18-positive cervical carcinoma cell line, which has been used to study different mechanisms of cervical cancer progression. An in vitro assay of scratch wound healing onto monolayers of SW756 cells was used to assess the effect of TNFalpha on cell migration into a wound space. It was found that SW756 cells have the ability to migrate, but not proliferate in response to scratch wounding in a serum-free medium supplemented with TNFalpha. RT-PCR analysis showed that SW756 cells express TNFalpha mRNA when incubated in medium with and without serum. Wound closure and migration rate of SW756 cells were significantly increased in the presence of serum-free media supplemented with TNFalpha (10 ng/mL) as compared to serum-free media, and media supplemented with either anti-TNFalpha antibody or both TNFalpha and anti-TNFalpha antibody (p < 0.05). The results showed a stimulatory effect of TNFalpha on the migration of SW756 cervical carcinoma cells, suggesting a novel and important role for TNFalpha in cervical cancer progression.
RESUMO
Birth is the result of complex, well-defined, and coordinated events, that are tightly regulated by endocrine, nervous, and immune responses, and take place primarily in the female reproductive tract. Various mechanisms and mediators involved in pregnancy, labor, and delivery, are highly conserved among different mammalian species and mast cells emerge as potential and crucial participants in these processes, as it is discussed in this review.
Assuntos
Humanos , Feminino , Gravidez , Mastócitos/metabolismo , Parto/fisiologia , Útero/metabolismo , Contração Muscular/fisiologia , Hormônio Liberador da Corticotropina/metabolismo , Hormônios Esteroides Gonadais/metabolismo , Mastócitos/citologia , Músculo Liso/fisiologia , Ocitocina/metabolismo , Útero/citologiaRESUMO
Birth is the result of complex, well-defined, and coordinated events, that are tightly regulated by endocrine, nervous, and immune responses, and take place primarily in the female reproductive tract. Various mechanisms and mediators involved in pregnancy, labor, and delivery, are highly conserved among different mammalian species and mast cells emerge as potential and crucial participants in these processes, as it is discussed in this review. (AU)
Assuntos
Humanos , Feminino , Gravidez , RESEARCH SUPPORT, NON-U.S. GOVT , Mastócitos/metabolismo , Parto/fisiologia , Útero/metabolismo , Hormônio Liberador da Corticotropina/metabolismo , Hormônios Esteroides Gonadais/metabolismo , Mastócitos/citologia , Contração Muscular/fisiologia , Músculo Liso/fisiologia , Ocitocina/metabolismo , Útero/citologiaRESUMO
The purpose of this review, based on studies from our laboratory as well as from others, is to summarize salient features of mast cell immunobiology and to describe their associations with gastrointestinal mucosal defense. Gastrointestinal mast cells are involved in many pathologic effects, such as food hypersensitivity. On the other hand, they also play a protective role in defense against parasitic and microbial infections. Thus, they have both positive and negative effects, but presently the mechanisms that control the balance of these various effects are poorly known. It has been suggested that stabilization of mast cells may be a key mechanism to protect the gastrointestinal tract from injury. Few molecules are known to possess both mast cell stabilizing and gastrointestinal cytoprotective activity. These include zinc compounds, sodium cromoglycate, FPL 52694, ketotifen, aloe vera, certain flavonoids such as quercetin, some sulfated proteoglycans such as chondroitin sulfate and dehydroleucodine. Dehydroleucodine, a sesquiterpene lactone isolated from Artemisia douglasiana Besser, exhibits anti-inflammatory and gastrointestinal cytoprotective action. The lactone stimulates mucus production, and inhibits histamine and serotonin release from intestinal mast cells. The lactone could act as a selective mast cell stabilizer by releasing cytoprotective factors and inhibiting pro-inflammatory mast cell mediators.
Assuntos
Humanos , Sistema Digestório , Mastócitos/citologia , Mastócitos/imunologia , Anti-Inflamatórios , Imunidade nas Mucosas/imunologia , Inflamação/tratamento farmacológico , Inflamação/imunologia , Inflamação/fisiopatologia , Lactonas/farmacologia , Lactonas/uso terapêutico , Mastócitos/efeitos dos fármacos , Mediadores da Inflamação/antagonistas & inibidores , Mediadores da Inflamação/metabolismo , Mucosa Gástrica/citologia , Mucosa Gástrica/efeitos dos fármacos , Mucosa Gástrica/metabolismo , Mucosa Intestinal/citologia , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/imunologia , Sesquiterpenos/farmacologia , Sesquiterpenos/uso terapêuticoRESUMO
The purpose of this review, based on studies from our laboratory as well as from others, is to summarize salient features of mast cell immunobiology and to describe their associations with gastrointestinal mucosal defense. Gastrointestinal mast cells are involved in many pathologic effects, such as food hypersensitivity. On the other hand, they also play a protective role in defense against parasitic and microbial infections. Thus, they have both positive and negative effects, but presently the mechanisms that control the balance of these various effects are poorly known. It has been suggested that stabilization of mast cells may be a key mechanism to protect the gastrointestinal tract from injury. Few molecules are known to possess both mast cell stabilizing and gastrointestinal cytoprotective activity. These include zinc compounds, sodium cromoglycate, FPL 52694, ketotifen, aloe vera, certain flavonoids such as quercetin, some sulfated proteoglycans such as chondroitin sulfate and dehydroleucodine. Dehydroleucodine, a sesquiterpene lactone isolated from Artemisia douglasiana Besser, exhibits anti-inflammatory and gastrointestinal cytoprotective action. The lactone stimulates mucus production, and inhibits histamine and serotonin release from intestinal mast cells. The lactone could act as a selective mast cell stabilizer by releasing cytoprotective factors and inhibiting pro-inflammatory mast cell mediators. (AU)
Assuntos
Humanos , Sistema Digestório/citologia , Sistema Digestório/imunologia , Mastócitos/citologia , Mastócitos/imunologia , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Mucosa Gástrica/citologia , Mucosa Gástrica/efeitos dos fármacos , Mucosa Gástrica/metabolismo , Imunidade nas Mucosas/imunologia , Inflamação/tratamento farmacológico , Inflamação/imunologia , Inflamação/fisiopatologia , Mediadores da Inflamação/antagonistas & inibidores , Mediadores da Inflamação/metabolismo , Mucosa Intestinal/citologia , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/imunologia , Lactonas/farmacologia , Lactonas/uso terapêutico , Mastócitos/efeitos dos fármacos , Sesquiterpenos/farmacologia , Sesquiterpenos/uso terapêuticoRESUMO
OBJECTIVE AND DESIGN: DhL, a lactone isolated from Artemisia douglasiana, prevents gastrointestinal damage elicited by necrosis-inducing agents and exhibits antiinflammatory action. This work examines the effect of DhL on compound 48/80-induced histamine and serotonin release in the isolated mouse jejunum, to determine whether DhL inhibits mediator release from mast cells at the enteric level. MATERIAL: Thirty jejuna from male Balb-c mice were used for the studies. TREATMENT: Samples were incubated sequentially in 9 test tubes containing RBS or 10 microg/ml compound 48/80 or 1.6 mmol/l + 10 microg/ml compound 48/80 at 37 degrees C for 90 minutes (10 min per tube). METHODS: Histamine and serotonin release studies, quantification of granulated mast cells, and evaluation of mast cell ultrastructure were carried out. Differences between groups were determined using analysis of variance followed by Tukey-Kramer multiple comparisons test. RESULTS: Compound 48/80 increased histamine and serotonin release by the tissue (141.95 +/- 62.58 pg/mg tissue vs basal 5.45 +/- 1.04, P<0.01 and 20.04 +/- 2.81 vs basal 9.24 +/- 1.56 ng/ mg tissue, P<0.01, respectively), decreased the number of granulated submucosal mast cells (0.077 +/- 0.0035 vs basal 0.14 +/- 0.015, P<0.05), and elicited evident granule ultrastructural changes. These effects were reduced by dehydroleucodine (19.51 +/- 7.88, P<0.01; 12.69 +/- 1, P<0.05 and 0.143 +/- 0.014, P<0.05, respectively). CONCLUSION: The lactone inhibits compound 48/80-induced histamine and serotonin release from mast cells in the isolated mouse jejunum.
Assuntos
Antiulcerosos/farmacologia , Histamina/metabolismo , Jejuno/metabolismo , Lactonas/farmacologia , Mastócitos/metabolismo , Serotonina/metabolismo , Sesquiterpenos/farmacologia , Animais , Antagonistas dos Receptores Histamínicos H1/farmacologia , Processamento de Imagem Assistida por Computador , Técnicas In Vitro , Jejuno/citologia , Jejuno/efeitos dos fármacos , Cetotifeno/farmacologia , Masculino , Mastócitos/efeitos dos fármacos , Mastócitos/ultraestrutura , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Eletrônica , Microscopia de Vídeo , p-Metoxi-N-metilfenetilamina/farmacologiaRESUMO
We have demonstrated that dehydroleucodine (DhL), a lactone isolated from Artemisia douglasiana Besser, prevents gastroduodenal damage induced by necrosis-inducing agents such as absolute ethanol. We have also reported, in a qualitative study, that this effect is related to the ability of the drug to stimulate mucus production. The present study was designed to quantitatively evaluate the effect of DhL on adherent mucus layer thickness, to obtain a more objective approach to the mechanism of action of the drug. Mice were divided into two groups: (I) controls were treated with orally administered carboxymethylcellulose (CMC) and (II) experimental animals received DhL in CMC. The thickness of the mucus gel layer was measured in unfixed stomachs and duodena, using an image analysis system. We observed an increase in the adherent mucus layer thickness in the experimental samples. This confirms that one of the main mechanisms involved in the cytoprotective action of the drug is mucus secretion.
Assuntos
Antiulcerosos/farmacologia , Mucosa Gástrica/efeitos dos fármacos , Mucosa Intestinal/efeitos dos fármacos , Lactonas/farmacologia , Muco/metabolismo , Sesquiterpenos/farmacologia , Animais , Mucosa Gástrica/metabolismo , Mucosa Intestinal/metabolismo , Masculino , CamundongosRESUMO
In previous work we have demonstrated that dehydroleucodine (DhL) prevents gastric damage induced by necrosis-inducing agents such as absolute ethanol (EtOH). In this study we examine the effects of DhL on gastroduodenal morphology and monoamine levels by histological and biochemical methods, respectively, as an approach to elucidating the cytoprotective mechanism of the drug. Histological evidence shows that DhL prevents formation of gastroduodenal mucosal lesions induced by EtOH and that this protective effect is related to the ability of the drug to stimulate mucus production. DhL itself does not affect the tissue concentration of NE, DA and 5-HT. However, it prevents the depletion of DA and 5-HT provoked by EtOH. We propose that the abundant mucoid blanket secreted after treatment with DhL acts as a diffusion barrier against EtOH. It is also possible that DhL could act as a "cell stabilizer," by inhibiting the degranulation of cells containing monoamines.