RESUMO
This study aimed to explore the mechanism of n-butanol alcohol extract of Baitouweng Decoction(BAEB) in the treatment of vulvovaginal candidiasis(VVC) in mice based on the negative regulation of NLRP3 inflammasome via PKCδ/NLRC4/IL-1Ra axis. In the experiment, female C57BL/6 mice were divided randomly into the following six groups: a blank control group, a VVC model group, high-, medium-, and low-dose BAEB groups(80, 40, and 20 mg·kg~(-1)), and a fluconazole group(20 mg·kg~(-1)). The VVC model was induced in mice except for those in the blank control group by the estrogen dependence method. After modeling, no treatment was carried out in the blank control group. The mice in the high-, medium-, and low-dose BAEB groups were treated with BAEB at 80, 40, and 20 mg·kg~(-1), respectively, and those in the fluconazole group were treated with fluconazole at 20 mg·kg~(-1). The mice in the VVC model group received the same volume of normal saline. The general state and body weight of mice in each group were observed every day, and the morphological changes of Candida albicans in the vaginal lavage of mice were examined by Gram staining. The fungal load in the vaginal lavage of mice was detected by microdilution assay. After the mice were killed, the degree of neutrophil infiltration in the vaginal lavage was detected by Papanicolaou staining. The content of inflammatory cytokines interleukin(IL)-1ß, IL-18, and lactate dehydrogenase(LDH) in the vaginal lavage was tested by enzyme-linked immunosorbent assay(ELISA), and vaginal histopathology was analyzed by hematoxylin-eosin(HE) staining. The expression and distribution of NLRP3, PKCδ, pNLRC4, and IL-1Ra in vaginal tissues were measured by immunohistochemistry(IHC), and the expression and distribution of pNLRC4 and IL-1Ra in vaginal tissues were detected by immunofluorescence(IF). The protein expression of NLRP3, PKCδ, pNLRC4, and IL-1Ra was detected by Western blot(WB), and the mRNA expression of NLRP3, PKCδ, pNLRC4, and IL-1Ra was detected by qRT-PCR. The results showed that compared with the blank control group, the VVC model group showed redness, edema, and white secretions in the vagina. Compared with the VVC model group, the BAEB groups showed improved general state of VVC mice. As revealed by Gram staining, Papanicolaou staining, microdilution assay, and HE staining, compared with the blank control group, the VVC model group showed a large number of hyphae, neutrophils infiltration, and increased fungal load in the vaginal lavage, destroyed vaginal mucosa, and infiltration of a large number of inflammatory cells. BAEB could reduce the transformation of C. albicans from yeast to hyphae. High-dose BAEB could significantly reduce neutrophil infiltration and fungal load. Low-and medium-dose BAEB could reduce the da-mage to the vaginal tissue, while high-dose BAEB could restore the damaged vaginal tissues to normal levels. ELISA results showed that the content of inflammatory cytokines IL-1ß, IL-18, and LDH in the VVC model group significantly increased compared with that in the blank control group, and the content of IL-1ß, IL-18 and LDH in the medium-and high-dose BAEB groups was significantly reduced compared with that in the VVC model group. WB and qRT-PCR results showed that compared with the blank control group, the VVC model group showed reduced protein and mRNA expression of PKCδ, pNLRC4, and IL-1Ra in vaginal tissues of mice and increased protein and mRNA expression of NLRP3. Compared with the VVC model group, the medium-and high-dose BAEB groups showed up-regulated protein and mRNA expression of PKCδ, pNLRC4, and IL-1Ra in vaginal tissues and inhibited protein and mRNA expression of NLRP3 in vaginal tissues. This study indicated that the therapeutic effect of BAEB on VVC mice was presumably related to the negative regulation of NLRP3 inflammasome by promoting PKCδ/NLRC4/IL-1Ra axis.
Assuntos
Candidíase Vulvovaginal , Medicamentos de Ervas Chinesas , Feminino , Animais , Humanos , Camundongos , Candidíase Vulvovaginal/tratamento farmacológico , Inflamassomos/genética , Interleucina-18 , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , 1-Butanol/farmacologia , Fluconazol/farmacologia , Fluconazol/uso terapêutico , Proteína Antagonista do Receptor de Interleucina 1/farmacologia , Proteína Antagonista do Receptor de Interleucina 1/uso terapêutico , Camundongos Endogâmicos C57BL , Candida albicans , Citocinas , Medicamentos de Ervas Chinesas/farmacologia , Etanol , RNA Mensageiro , Proteínas de Ligação ao Cálcio/farmacologia , Proteínas de Ligação ao Cálcio/uso terapêuticoRESUMO
This study aimed to explore the mechanism of n-butanol alcohol extract of Baitouweng Decoction(BAEB) in the treatment of vulvovaginal candidiasis(VVC) in mice based on the negative regulation of NLRP3 inflammasome via PKCδ/NLRC4/IL-1Ra axis. In the experiment, female C57BL/6 mice were divided randomly into the following six groups: a blank control group, a VVC model group, high-, medium-, and low-dose BAEB groups(80, 40, and 20 mg·kg~(-1)), and a fluconazole group(20 mg·kg~(-1)). The VVC model was induced in mice except for those in the blank control group by the estrogen dependence method. After modeling, no treatment was carried out in the blank control group. The mice in the high-, medium-, and low-dose BAEB groups were treated with BAEB at 80, 40, and 20 mg·kg~(-1), respectively, and those in the fluconazole group were treated with fluconazole at 20 mg·kg~(-1). The mice in the VVC model group received the same volume of normal saline. The general state and body weight of mice in each group were observed every day, and the morphological changes of Candida albicans in the vaginal lavage of mice were examined by Gram staining. The fungal load in the vaginal lavage of mice was detected by microdilution assay. After the mice were killed, the degree of neutrophil infiltration in the vaginal lavage was detected by Papanicolaou staining. The content of inflammatory cytokines interleukin(IL)-1β, IL-18, and lactate dehydrogenase(LDH) in the vaginal lavage was tested by enzyme-linked immunosorbent assay(ELISA), and vaginal histopathology was analyzed by hematoxylin-eosin(HE) staining. The expression and distribution of NLRP3, PKCδ, pNLRC4, and IL-1Ra in vaginal tissues were measured by immunohistochemistry(IHC), and the expression and distribution of pNLRC4 and IL-1Ra in vaginal tissues were detected by immunofluorescence(IF). The protein expression of NLRP3, PKCδ, pNLRC4, and IL-1Ra was detected by Western blot(WB), and the mRNA expression of NLRP3, PKCδ, pNLRC4, and IL-1Ra was detected by qRT-PCR. The results showed that compared with the blank control group, the VVC model group showed redness, edema, and white secretions in the vagina. Compared with the VVC model group, the BAEB groups showed improved general state of VVC mice. As revealed by Gram staining, Papanicolaou staining, microdilution assay, and HE staining, compared with the blank control group, the VVC model group showed a large number of hyphae, neutrophils infiltration, and increased fungal load in the vaginal lavage, destroyed vaginal mucosa, and infiltration of a large number of inflammatory cells. BAEB could reduce the transformation of C. albicans from yeast to hyphae. High-dose BAEB could significantly reduce neutrophil infiltration and fungal load. Low-and medium-dose BAEB could reduce the da-mage to the vaginal tissue, while high-dose BAEB could restore the damaged vaginal tissues to normal levels. ELISA results showed that the content of inflammatory cytokines IL-1β, IL-18, and LDH in the VVC model group significantly increased compared with that in the blank control group, and the content of IL-1β, IL-18 and LDH in the medium-and high-dose BAEB groups was significantly reduced compared with that in the VVC model group. WB and qRT-PCR results showed that compared with the blank control group, the VVC model group showed reduced protein and mRNA expression of PKCδ, pNLRC4, and IL-1Ra in vaginal tissues of mice and increased protein and mRNA expression of NLRP3. Compared with the VVC model group, the medium-and high-dose BAEB groups showed up-regulated protein and mRNA expression of PKCδ, pNLRC4, and IL-1Ra in vaginal tissues and inhibited protein and mRNA expression of NLRP3 in vaginal tissues. This study indicated that the therapeutic effect of BAEB on VVC mice was presumably related to the negative regulation of NLRP3 inflammasome by promoting PKCδ/NLRC4/IL-1Ra axis.
Assuntos
Feminino , Animais , Humanos , Camundongos , Candidíase Vulvovaginal/tratamento farmacológico , Inflamassomos/genética , Interleucina-18 , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , 1-Butanol/farmacologia , Fluconazol/uso terapêutico , Proteína Antagonista do Receptor de Interleucina 1/uso terapêutico , Camundongos Endogâmicos C57BL , Candida albicans , Citocinas , Medicamentos de Ervas Chinesas/farmacologia , Etanol , RNA Mensageiro , Proteínas de Ligação ao Cálcio/uso terapêuticoRESUMO
Polcalcins are important respiratory panallergens, whose IgE-binding capacity depends on the presence of calcium. Since specific immunotherapy is not yet available for the treatment of polcalcin-sensitized patients, we aimed to develop a molecule for efficient and safe immunotherapy. We generated a hypoallergenic variant of the grass pollen polcalcin Phl p 7 by introducing specific point mutations into the allergen's calcium-binding regions. We thereby followed a mutation strategy that had previously resulted in a hypoallergenic mutant of a calcium-binding food allergen, the major fish allergen parvalbumin. Dot blot assays performed with sera from Phl p 7-sensitized patients showed a drastically reduced IgE reactivity of the Phl p 7 mutant in comparison to wildtype Phl p 7, and basophil activation assays indicated a significantly reduced allergenic activity. Rabbit IgG directed against mutant rPhl p 7 blocked patients' IgE binding to wildtype Phl p 7, indicating the mutant's potential applicability for immunotherapy. Mass spectrometry and circular dichroism experiments showed that the mutant had lost the calcium-binding capacity, but still represented a folded protein. In silico analyses revealed that the hypoallergenicity might be due to fewer negative charges on the molecule's surface and an increased molecular flexibility. We thus generated a hypoallergenic Phl p 7 variant that could be used for immunotherapy of polcalcin-sensitized individuals.
Assuntos
Antígenos de Plantas/uso terapêutico , Proteínas de Ligação ao Cálcio/uso terapêutico , Poaceae/imunologia , Pólen/imunologia , Rinite Alérgica Sazonal/terapia , Animais , Antígenos de Plantas/genética , Antígenos de Plantas/imunologia , Proteínas de Ligação ao Cálcio/genética , Proteínas de Ligação ao Cálcio/imunologia , Feminino , Humanos , Imunoglobulina E/imunologia , Imunoterapia , Masculino , Modelos Moleculares , Mutação Puntual , Engenharia de Proteínas , Coelhos , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/uso terapêutico , Rinite Alérgica Sazonal/imunologiaRESUMO
Vascular calcification was once regarded as an advanced stage of atherosclerosis only. However, calcification is currently considered as highly regulated and potentially reversible process.Matrix Gla protein (MGP) represents natural inhibitor of vascular calcification, whereas vitamin K is key co-factor of its maturation to the active form. There is accumulating evidence that vitamin K status and corresponding MGP activity may influence cardiovascular risk. This review summarizes pathophysiological mechanism and recent evidence relative to MGP. Moreover, available data concerning vitamin K supplementation are depicted.
Assuntos
Proteínas de Ligação ao Cálcio/uso terapêutico , Proteínas da Matriz Extracelular/uso terapêutico , Calcificação Vascular/prevenção & controle , Vitamina K/uso terapêutico , Antifibrinolíticos/uso terapêutico , Suplementos Nutricionais , Humanos , Fatores de Risco , Proteína de Matriz GlaRESUMO
The major turnip (Brassica rapa) pollen allergen, belongs to a family of calcium-binding proteins (i.e., two EF-hand proteins), which occur as highly cross-reactive allergens in pollen of weeds, grasses and trees. In this study, the IgE binding capacity and allergenic activity of three recombinant allergen variants containing mutations in their calcium-binding sites were analyzed in sensitized patients with the aim to identify the most suitable hypoallergenic molecule for specific immunotherapy. Analysis of the wildtype allergen and the mutants regarding IgE reactivity and activation of basophils in allergic patients indicated that the allergen derivative mutated in both calcium-binding domains had the lowest allergenic activity. Gel filtration and circular dichroism experiments showed that both, the wildtype and the double mutant, occurred as dimers in solution and assumed alpha-helical fold, respectively. However, both fold and thermal stability were considerably reduced in the double mutant. The use of bioinformatic tools for evaluation of the solvent accessibility and charge distribution suggested that the reduced IgE reactivity and different structural properties of the double mutant may be due to a loss of negatively charged amino acids on the surface. Interestingly, immunization of rabbits showed that only the double mutant but not the wildtype allergen induced IgG antibodies which recognized the allergen and blocked binding of allergic patients IgE. Due to the extensive structural similarity and cross-reactivity between calcium-binding pollen allergens the hypoallergenic double mutant may be useful not only for immunotherapy of turnip pollen allergy, but also for the treatment of allergies to other two EF-hand pollen allergens.
Assuntos
Basófilos/efeitos dos fármacos , Brassica rapa/imunologia , Proteínas de Ligação ao Cálcio/imunologia , Proteínas de Ligação ao Cálcio/uso terapêutico , Dessensibilização Imunológica/métodos , Proteínas de Plantas/imunologia , Proteínas de Plantas/uso terapêutico , Rinite Alérgica Sazonal/terapia , Adulto , Alérgenos/genética , Alérgenos/imunologia , Alérgenos/uso terapêutico , Sequência de Aminoácidos , Animais , Formação de Anticorpos/efeitos dos fármacos , Antígenos de Plantas/genética , Antígenos de Plantas/uso terapêutico , Basófilos/imunologia , Proteínas de Ligação ao Cálcio/genética , Degranulação Celular/efeitos dos fármacos , Células Cultivadas , Reações Cruzadas , Feminino , Humanos , Imunoglobulina E/metabolismo , Masculino , Dados de Sequência Molecular , Mutação/genética , Proteínas de Plantas/genética , Pólen/efeitos adversos , Pólen/imunologia , Conformação Proteica , Engenharia de Proteínas , Coelhos , Rinite Alérgica Sazonal/imunologia , Adulto JovemRESUMO
A most common and trusted source of Ca is milk or other dairy products. However, some oriental people do not drink milk due to lactose indigestion and intolerance, which make them allergic to milk. There have been many studies on alternative calcium-rich diet or Ca supplements. Among them, teleost fish like anchovy and mola, which are commonly consumed in Asian countries, could be an important Ca dietary supplement, especially in population groups with low intakes of milk and dairy products. In this chapter, we summarize beneficial effects of teleost fish bone peptide (FBP) for Ca bioavailability and bone mineralization, based on our researches.
Assuntos
Conservadores da Densidade Óssea/uso terapêutico , Osso e Ossos/metabolismo , Proteínas de Ligação ao Cálcio/uso terapêutico , Suplementos Nutricionais , Proteínas de Peixes/uso terapêutico , Peixes/metabolismo , Fragmentos de Peptídeos/uso terapêutico , Animais , Conservadores da Densidade Óssea/isolamento & purificação , Conservadores da Densidade Óssea/metabolismo , Cálcio da Dieta/administração & dosagem , Cálcio da Dieta/metabolismo , Cálcio da Dieta/uso terapêutico , Proteínas de Ligação ao Cálcio/isolamento & purificação , Proteínas de Ligação ao Cálcio/metabolismo , Proteínas de Peixes/isolamento & purificação , Proteínas de Peixes/metabolismo , Humanos , Absorção Intestinal , Osteogênese , Fragmentos de Peptídeos/isolamento & purificação , Fragmentos de Peptídeos/metabolismoRESUMO
Distinct Notch ligands possess a characteristic ability in terms of functional T cell differentiation. However, the precise role or the therapeutic potential of each Notch ligand in autoimmune diseases is largely unknown. In this study, we examined whether Jagged1 modulates a collagen-induced rheumatoid arthritis (CIA) model by altering T cell responses. The injection of a soluble Jagged1-encoding plasmid, sJag1-P, before or even after initial type II collagen (CII) immunization suppressed the disease severity of CIA. However, this treatment did not suppress CII-specific CD4(+) T cell proliferation and CII-specific Ab production. Depletion of either CD4(+) or CD8(+) T cells ameliorated CIA severity and sJag1-P further improved CIA in CD4(+) but not CD8(+) T cell-depleted mice. Injection of OVA and Jagged1-encoding plasmids inhibited proliferation of OVA-specific granzyme B-producing CD8(+) T cells, although Jagged1 could not directly inhibit CD8(+) T cell proliferation in vitro. The blockade of Jagged1 by an anti-Jagged1 Ab exacerbated CIA, whereas this effect was not observed in the absence of CD8(+) T cells. These data indicate that Jagged1 is able to deliver an indirect negative signal into CD8(+) T cells in vivo, which suggests its therapeutic potential in the treatment of CD8(+) T cell-mediated diseases, including rheumatoid arthritis.
Assuntos
Artrite Experimental/imunologia , Artrite Experimental/terapia , Linfócitos T CD8-Positivos/imunologia , Proteínas de Ligação ao Cálcio/fisiologia , Proteínas de Ligação ao Cálcio/uso terapêutico , Inibidores do Crescimento/uso terapêutico , Peptídeos e Proteínas de Sinalização Intercelular/fisiologia , Peptídeos e Proteínas de Sinalização Intercelular/uso terapêutico , Ativação Linfocitária/imunologia , Proteínas de Membrana/fisiologia , Proteínas de Membrana/uso terapêutico , Transdução de Sinais/imunologia , Animais , Artrite Experimental/patologia , Linfócitos T CD8-Positivos/patologia , Proteínas de Ligação ao Cálcio/administração & dosagem , Proteínas de Ligação ao Cálcio/genética , Linhagem Celular , Células Cultivadas , Regulação para Baixo/genética , Regulação para Baixo/imunologia , Inibidores do Crescimento/administração & dosagem , Inibidores do Crescimento/genética , Inibidores do Crescimento/fisiologia , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/administração & dosagem , Peptídeos e Proteínas de Sinalização Intercelular/genética , Proteína Jagged-1 , Células Jurkat , Ativação Linfocitária/genética , Depleção Linfocítica , Masculino , Proteínas de Membrana/administração & dosagem , Proteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos DBA , Plasmídeos/administração & dosagem , Plasmídeos/genética , Plasmídeos/uso terapêutico , Proteínas Serrate-Jagged , Transdução de Sinais/genética , Vacinas de DNA/administração & dosagem , Vacinas de DNA/genética , Vacinas de DNA/uso terapêuticoRESUMO
Human healthy skin is continuously exposed to bacteria, but is particularly resistant to the common gut bacterium Escherichia coli. We show here that keratinocytes secrete, as the main E. coli-killing compound, the S100 protein psoriasin in vitro and in vivo in a site-dependent way. In vivo treatment of human skin with antibodies to psoriasin inhibited its E. coli-killing properties. Psoriasin was induced in keratinocytes in vitro and in vivo by E. coli, indicating that its focal expression in skin may derive from local microbial induction. Zn(2+)-saturated psoriasin showed diminished antimicrobial activity, suggesting that Zn(2+) sequestration could be a possible antimicrobial mechanism. Thus, psoriasin may be key to the resistance of skin against E. coli.
Assuntos
Antibacterianos/uso terapêutico , Proteínas de Ligação ao Cálcio/uso terapêutico , Infecções por Escherichia coli/prevenção & controle , Escherichia coli/efeitos dos fármacos , Antibacterianos/farmacologia , Proteínas de Ligação ao Cálcio/farmacologia , Escherichia coli/metabolismo , Humanos , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , Proteína A7 Ligante de Cálcio S100 , Proteínas S100 , Pele/microbiologia , Zinco/metabolismo , Zinco/farmacologiaRESUMO
Cardiovascular disease is the largest cause of mortality in hemodialysis patients. Cardiovascular mortality is fivefold to twentyfold higher in hemodialysis patients than in the general population. Atherosclerosis and vascular calcification are the characteristic complications in hemodialysis patients. Hemodialysis patients have traditional risk factors such as abnormal lipid metabolism and uremia-related risk factors such as oxidative stress and hyperphosphatemia. Oxidative stress takes place by increased production of oxidants by leukocytes and antioxidant loss of vitamin C and E. Oxidatively modified LDL exist in the circulation by excess of oxidative stress in hemodialysis patients. Oxidative stress is a major contributor to accelerated development atherosclerosis. Oxidative stress and hyperphosphatemia also influence vascular calcification. The pattern of vascular calcification in hemodialysis patient is characterized by mineral deposition in the tunica media. It is reported that the obvious calcification in aorta and artery of the MGP knockout mouse is recognized. It is indicated that MGP has the inhibitory effect of the calcification of vessel wall. Vitamin E protects atherosclerosis and vascular calcification in hemodialysis patients. It is also important to control hyperphosphatemia for vascular calcification.
Assuntos
Arteriosclerose/etiologia , Calcinose/etiologia , Doenças Cardiovasculares/etiologia , Diálise Renal/efeitos adversos , Animais , Arteriosclerose/prevenção & controle , Calcinose/prevenção & controle , Proteínas de Ligação ao Cálcio/fisiologia , Proteínas de Ligação ao Cálcio/uso terapêutico , Doenças Cardiovasculares/prevenção & controle , Causas de Morte , Proteínas da Matriz Extracelular/fisiologia , Proteínas da Matriz Extracelular/uso terapêutico , Humanos , Metabolismo dos Lipídeos , Doenças Metabólicas/complicações , Camundongos , Estresse Oxidativo/fisiologia , Fósforo/sangue , Distúrbios do Metabolismo do Fósforo/complicações , Diálise Renal/mortalidade , Fatores de Risco , Uremia/complicações , Vitamina E/uso terapêutico , Proteína de Matriz GlaRESUMO
Tumor growth depends upon an adequate supply of oxygen and nutrients achieved through angiogenesis and maintenance of an intact tumor vasculature. Therapy with individual agents that target new vessel formation or existing vessels has suppressed experimental tumor growth, but rarely resulted in the eradication of tumors. We therefore tested the combined anti-tumor activity of vasostatin and interferon-inducible protein-10 (IP-10), agents that differently target the tumor vasculature. Vasostatin, a selective and direct inhibitor of endothelial cell proliferation, significantly reduced Burkitt tumor growth and tumor vessel density. IP-10, an "angiotoxic" chemokine, caused vascular damage and focal necrosis in Burkitt tumors. When combined, vasostatin plus IP-10 reduced tumor growth more effectively than each agent alone, but complete tumor regression was not observed. Microscopically, these tumors displayed focal necrosis and reduction in vessel density. Combination therapy with the inhibitors of angiogenesis vasostatin and IP-10 is effective in reducing the rate of tumor growth but fails to induce tumor regression, suggesting that curative treatment may require supplemental drugs targeting directly the tumor cells.
Assuntos
Inibidores da Angiogênese/uso terapêutico , Antineoplásicos/uso terapêutico , Linfoma de Burkitt/tratamento farmacológico , Proteínas de Ligação ao Cálcio/uso terapêutico , Quimiocina CXCL10/uso terapêutico , Neovascularização Patológica/tratamento farmacológico , Fragmentos de Peptídeos/uso terapêutico , Ribonucleoproteínas/uso terapêutico , Actinas/análise , Animais , Apoptose/efeitos dos fármacos , Linfoma de Burkitt/prevenção & controle , Calreticulina , Divisão Celular/efeitos dos fármacos , Avaliação Pré-Clínica de Medicamentos , Sinergismo Farmacológico , Endotélio Vascular/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Músculo Liso Vascular/efeitos dos fármacos , Molécula-1 de Adesão Celular Endotelial a Plaquetas/análise , Proteínas Recombinantes de Fusão/uso terapêutico , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Calponin h1 (CNh1) is an actin-binding protein that is expressed mainly in smooth muscle cells and is known to regulate smooth muscle contraction. Recently, re-expression of CNh1 in leiomyosarcoma cell lines is reported to suppress cell proliferation and tumorigenicity. However, little is known about the associated cellular structural and functional changes. Since CNh1 is also detected in normal fibroblasts, we hypothesised that CNh1 would also inhibit cell proliferation of the fibrosarcoma cells, HT1080, in which CNh1 is suppressed. An expression vector of human CNh1 complementary DNA was transfected into human HT1080 cells by a calcium-phosphate precipitation method. CNh1-transfected cells exhibited a flattened morphology with organised actin filaments, a significant decrease in cell motility and enhancement in adhesion to fibronectin in association with an increase in integrin alpha5beta1 expression. Anchorage-independent growth and tumorigenicity in nude mice were suppressed in the CNh1-transfected cells. Our results suggest that CNh1 may have a role as a tumour suppressor in human fibrosarcoma by influencing cytoskeletal activities.
Assuntos
Proteínas de Ligação ao Cálcio/uso terapêutico , Fibrossarcoma/tratamento farmacológico , Animais , Divisão Celular/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Fibrossarcoma/patologia , Humanos , Imuno-Histoquímica , Camundongos , Camundongos Endogâmicos BALB C , Proteínas dos Microfilamentos , Transplante de Neoplasias , Células Tumorais Cultivadas , CalponinasRESUMO
Oral linomide, (quinoline-3-carboxamide), has been shown to prevent autoimmune insulitis, islet destruction, and diabetes in NOD mice treated at an early stage of the disease, but confers only partial protection in animals with advanced disease. Reg protein, the gene product of a complementary DNA isolated from a regenerating rat islet library, has been previously shown to induce expansion of beta-cell mass in pancreatectomized rats. To determine the effect of treatment combining immunomodulation and Reg protein on advanced autoimmune diabetes, we treated female NOD mice with oral linomide and i.p. Reg protein injections. In 14-week-old animals with less severe disease (glucose tolerant), treatment with each agent alone resulted in amelioration of diabetes, as did treatment with Reg alone in 5-week-old prediabetic mice. In 14-week-old animals with more severe disease (glucose intolerant), only treatment with the combination of both agents, but not that with each separately, resulted in amelioration of diabetes. Our study suggests that treatment aimed at abrogation of autoimmunity combined with expansion of beta-cell mass constitutes a potential therapeutic approach for treatment of insulin-dependent diabetes mellitus.
Assuntos
Adjuvantes Imunológicos/uso terapêutico , Proteínas de Ligação ao Cálcio/uso terapêutico , Diabetes Mellitus Tipo 1/terapia , Hidroxiquinolinas/uso terapêutico , Imunoterapia , Proteínas do Tecido Nervoso , Animais , Doenças Autoimunes , Glicemia/metabolismo , Proteínas de Ligação ao Cálcio/administração & dosagem , Diabetes Mellitus Tipo 1/imunologia , Diabetes Mellitus Tipo 1/patologia , Feminino , Teste de Tolerância a Glucose , Hidroxiquinolinas/administração & dosagem , Insulina/análise , Ilhotas Pancreáticas/patologia , Litostatina , Camundongos , Camundongos Endogâmicos NOD , Pâncreas/químicaAssuntos
Edema Encefálico/prevenção & controle , Isquemia Encefálica/complicações , Dexametasona/uso terapêutico , Animais , Anexinas , Barreira Hematoencefálica/efeitos dos fármacos , Edema Encefálico/etiologia , Proteínas de Ligação ao Cálcio/uso terapêutico , Gatos , Dexametasona/administração & dosagem , Esquema de Medicação , Manitol/uso terapêuticoRESUMO
Transport features of calcium in red cells of rats with spontaneous genetic hypertension (SHR) have been studied. It is shown that in the presence of calmodulin the rate of calcium accumulation by the inside-out vesicles of SHR red cell membranes is roughly twice less than in the control normotensive rats. It is suggested that upset interrelationship between calmodulin and Mg-Ca-ATPase of plasma membrane may be the cause of increase of intracellular calcium recorded in a number of tissues in primary hypertension.