RESUMO
We developed a drug delivery system for atherosclerotic lesions using immuno-liposomes. We focused on enhancing the delivery efficiency of the liposomes to macrophages in atherosclerotic lesions by antibody modification of lectinlike oxidized low-density lipoproteins (LDL) receptor 1 (LOX-1). The cellular accumulation of the liposomes in foam cells induced by oxidized LDL (oxLDL) in Raw264 mouse macrophages was evaluated. The cellular accumulation of LOX-1 antibody modified liposomes in oxLDL-induced foam cells and untreated Raw264 cells was significantly higher compared with that of unmodified liposomes. The liposomes were also administered intravenously to Apoeshl mice as an atherosclerosis model. Frozen sections were prepared from the mouse aortas and observed by confocal laser microscopy. The distribution of LOX-1 antibody modified liposomes in the atherosclerotic regions of Apoeshl mice was significantly greater compared with that of unmodified liposomes. The results suggest that LOX-1 antibody modified liposomes can target foam cells in atherosclerotic lesions, providing a potential route for delivering various drugs with pharmacological effects or detecting atherosclerotic foci for the diagnosis of atherosclerosis.
Assuntos
Aterosclerose , Lipossomos , Animais , Camundongos , Portadores de Fármacos , Anticorpos , Macrófagos , Apolipoproteínas E , Aterosclerose/tratamento farmacológico , Receptores Depuradores Classe ERESUMO
Objective: We aimed to investigate the involvement of efflux transporters, including multidrug resistant protein 1 (MDR1), multidrug resistance-associated protein 1 (MRP1), MRP2, and breast cancer resistance protein (BCRP), in the intracellular accumulation of the antifibrotic agent nintedanib in fibrotic lung cells. Methods: We used transforming growth factor-ß1 (TGF-ß1)-treated human lung fibroblasts (WI-38) and alveolar epithelial cells (A549) as in vitro models. The expression and activities of efflux transporters in TGF-ß1-treated WI-38 and A549 cells were evaluated using immunoblotting and flow cytometry. Cells were treated with nintedanib and then incubated with inhibitors of these transporters. The intracellular concentration of nintedanib was determined. Results: MDR1, MRP1, MRP2, and BCRP were found to be expressed in WI-38 and A549 cells with or without TGF-ß1 stimulation, with the exception of MRP2 in WI-38 cells. The efflux activities of these transporters were observed in these cells. MDR1 inhibitors significantly increased the intracellular accumulation of nintedanib, whereas MRP inhibitors did not show an effect. The BCRP inhibitor significantly increased the transporter activity in A549 cells but not in WI-38 cells. Conclusion: This study suggests that the efflux via MDR1 and BCRP is involved in the intracellular accumulation of nintedanib in fibrotic lung cells.
Assuntos
Fibrose Pulmonar Idiopática/tratamento farmacológico , Indóis/farmacocinética , Pulmão/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Células A549 , Células Epiteliais Alveolares/metabolismo , Linhagem Celular , Fibroblastos/metabolismo , Humanos , Indóis/administração & dosagem , Pulmão/citologia , Inibidores de Proteínas Quinases/administração & dosagem , Inibidores de Proteínas Quinases/farmacocinética , Distribuição Tecidual , Fator de Crescimento Transformador beta1/administração & dosagemRESUMO
We have shown that clarithromycin (CAM), a macrolide antibiotic, more highly distributes from plasma to lung epithelium lining fluid (ELF), the infection site of pathogens, than azithromycin (AZM) and telithromycin (TEL). Transporter(s) expressed on lung epithelial cells may contribute to the distribution of the compiunds to the ELF. However, distribution mechanisms are not well known. In this study, their transport characteristics in Calu-3 cell monolayers as model lung epithelial cells were examined. The basolateral-to-apical transport of CAM through Calu-3 cell monolayers was greater than that of AZM and TEL. Although verapamil and cyclosporine A as MDR1 substrates completely inhibited the basolateral-to-apical transport, probenecid as MRP1 inhibitor did not show an effect. These results suggest that the antibiotics are transported from plasma to ELF by MDR1 of lung epithelial cells. In addition, their affinity and binding rate to MDR1 was examined by ATP activity assay. The affinity and binding rate of CAM was greater than those of AZM and TEL. These corresponded with the distributions from plasma to ELF as described above. The present study suggests that the more highly distribution of CAM from plasma to ELF is due to the high affinity and binding rate to MDR1 of lung epithelial cells.
Assuntos
Antibacterianos/metabolismo , Azitromicina/farmacocinética , Claritromicina/farmacocinética , Células Epiteliais/metabolismo , Cetolídeos/farmacocinética , Pulmão/metabolismo , Infecções Respiratórias/tratamento farmacológico , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/antagonistas & inibidores , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Adenosina Trifosfatases/metabolismo , Algoritmos , Animais , Área Sob a Curva , Transporte Biológico Ativo/efeitos dos fármacos , Linhagem Celular , Pulmão/citologia , Camundongos , Proteínas Associadas à Resistência a Múltiplos Medicamentos/antagonistas & inibidores , Proteínas Associadas à Resistência a Múltiplos Medicamentos/metabolismo , Ligação ProteicaRESUMO
Sperminated dextrans (SD) having different average molecular weights (MWs; 10, 40 and 70 kDa) and numbers of amino groups were prepared as cationized polymers for use as absorption enhancers. The absorption enhancing effects on the pulmonary absorption of insulin in rats and the permeation of FITC-dextran (MW 4,400, FD4) through calu-3 cell (human airway epithelial cell) monolayers by SD were evaluated. SD significantly enhanced the pulmonary absorption of insulin SD and the permeation of FD4 through calu-3 cells. The enhancing effects on the absorption insulin and permeation of FD4 through calu-3 cells increased with an increase in the molecular weigh of SD over the range 10-70 kDa. SD may interact directly with the luminal surface of mucus membranes via an ion-ion interaction and then induce signals that open tight junctions resulting in intercellular permeation of water soluble drugs. SD may be useful as an absorption enhancer for pulmonary delivery of peptide and protein drugs.
Assuntos
Dextranos/farmacologia , Excipientes/farmacologia , Pulmão/metabolismo , Absorção/efeitos dos fármacos , Administração por Inalação , Aminas/química , Animais , Glicemia/metabolismo , Linhagem Celular , Permeabilidade da Membrana Celular/efeitos dos fármacos , Dextranos/síntese química , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Excipientes/síntese química , Fluoresceína-5-Isotiocianato/análogos & derivados , Fluoresceína-5-Isotiocianato/farmacologia , Humanos , Hipoglicemiantes/administração & dosagem , Hipoglicemiantes/farmacocinética , Insulina/administração & dosagem , Insulina/farmacocinética , Pulmão/efeitos dos fármacos , Masculino , Peso Molecular , Peptídeos/administração & dosagem , Peptídeos/farmacocinética , Permeabilidade , Ratos , Ratos Wistar , Espermina/análogos & derivados , Espermina/síntese químicaRESUMO
Improving the quality of ulcer healing (QOUH) is one of the valid methods of prevention of relapse of gastric ulcers. We investigated the effect of lafutidine on the QOUH of gastric ulcer compared with famotidine in a randomized, multi-centre controlled trial. Consecutive 80 patients with a gastric ulcer were randomly assigned to receive twice daily either lafutidine (10 mg) or famotidine (20 mg) for 12 weeks. Esophagogastroduodenoscopy was performed to examine the ulcer healing rate and rate of flat type ulcer scars using dye-contrast. The gastric ulcer healing rate was 92.1% in the lafutidine group (35/38) and 94.7% in the famotidine group (36/38). The rate of flat-type ulcer scars was significantly higher in the lafutidine group (68.4%, 26/38) than in the famotidine group (42.1%, 16/38) (P = 0.021). In conclusion, the present study demonstrated that lafutidine, as compared to famotidine, yields a significantly superior QOUH in patients with gastric ulcers in the clinical setting.
Assuntos
Acetamidas/uso terapêutico , Antiulcerosos/uso terapêutico , Piperidinas/uso terapêutico , Piridinas/uso terapêutico , Úlcera Gástrica/tratamento farmacológico , Acetamidas/administração & dosagem , Adulto , Idoso , Antiulcerosos/administração & dosagem , Esquema de Medicação , Famotidina/administração & dosagem , Famotidina/uso terapêutico , Feminino , Gastroscopia , Humanos , Masculino , Pessoa de Meia-Idade , Piperidinas/administração & dosagem , Piridinas/administração & dosagem , Úlcera Gástrica/diagnóstico , Resultado do Tratamento , Cicatrização/efeitos dos fármacosRESUMO
Ranitidine has been found to have anti-inflammatory action as well as antisecretory action in experimental models. However, there are no reports in human gastric ulcer. The aim of this study was to investigate the effects of ranitidine compared with those of famotidine on the quality of gastric ulcer healing. We randomly assigned 69 consecutive patients with gastric ulcers to ranitidine (n = 34) or famotidine (n = 35) for 12 weeks, with endoscopic assessment of the quality of gastric ulcer healing and histological assessment of gastric mucosa 12 weeks after treatment started. Ulcer healing rates of over 95% were very similar in the two groups. The rates of ulcer scars with a flat pattern (good-quality healing) were significantly higher in the ranitidine group than in the famotidine group (per protocol, 63.0% and 34.5%, p = 0.033). The neutrophil infiltration score in the body mucosa treated with famotidine, but not ranitidine, significantly increased after treatment. In contrast, the mononuclear cell infiltration score in the antral mucosa treated with ranitidine, but not in that treated with famotidine, had significantly decreased. In conclusion, initial therapy with ranitidine significantly improved the quality of gastric ulcer healing and the histological scores of gastric mucosa compared with famotidine.
Assuntos
Antiulcerosos/uso terapêutico , Famotidina/uso terapêutico , Antagonistas dos Receptores H2 da Histamina/uso terapêutico , Ranitidina/uso terapêutico , Úlcera Gástrica/tratamento farmacológico , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Mucosa Gástrica/efeitos dos fármacos , Mucosa Gástrica/imunologia , Mucosa Gástrica/patologia , Gastroscopia , Humanos , Contagem de Leucócitos , Masculino , Pessoa de Meia-Idade , Cicatrização/efeitos dos fármacosRESUMO
The mortality rate of recurrent esophageal carcinoma remains high because of its resistance to chemotherapy and radiation therapy. We present a patient with recurrent esophageal carcinoma, which dramatically disappeared after treatment with the combination of continuous infusion of 5-fluorouracil and low-dose cis-Diamminedichloroplatinum-II (cisplatin) infusion (FP therapy). Furthermore, we immunohistologically found that glutathione S-transferases (GST)-pi, a marker of resistance to cisplatin, was faintly expressed both in the endoscopical biopsy specimens of recurrent tumor and in the resected specimens of esophageal carcinoma and metastatic lymph nodes. FP therapy was suggested to be effective for recurrent esophageal carcinoma. Immunostaining for GST-pi might be a prospective marker for the sensitivity of esophageal carcinoma to FP therapy, particularly cisplatin.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Carcinoma/tratamento farmacológico , Carcinoma/enzimologia , Neoplasias Esofágicas/tratamento farmacológico , Neoplasias Esofágicas/enzimologia , Glutationa Transferase/metabolismo , Isoenzimas/metabolismo , Recidiva Local de Neoplasia/patologia , Antimetabólitos Antineoplásicos/administração & dosagem , Antineoplásicos/administração & dosagem , Carcinoma/diagnóstico por imagem , Carcinoma/patologia , Cisplatino/administração & dosagem , Relação Dose-Resposta a Droga , Neoplasias Esofágicas/diagnóstico por imagem , Neoplasias Esofágicas/patologia , Feminino , Fluoruracila/administração & dosagem , Glutationa S-Transferase pi , Humanos , Bombas de Infusão , Pessoa de Meia-Idade , Tomografia Computadorizada por Raios XRESUMO
For the purpose to investigate the immunological anti-tumor function of lymphocytes in the regional lymph nodes of gastric cancer, a double staining procedure using several monoclonal antibodies against the surface membranes of T and NK cells with fluorescent antibody technique was conducted. The number of CD11b+ cells out of CD8+ cells was small with almost all being CD11b- belonging to cytotoxic T cells. Leu8+ cells and Leu8- cells out of CD4+ cells were recognized in equal numbers and were reciprocal. The ratio of CD4+ Leu8-/CD4+ demonstrated an increase in the group injected with OK-432. The ratio of OKT9+ or OKIa1+ occupied in CD8+ or CD4+ cells was only several percent, and few in number. An increase for the ratio of CD4+ OKT9+/CD4+ was observed in the group injected with IL-2. Similar increases for the ratio of CD4+ OKIa1+/CD4+ were obtained in the group injected with OK-432 and in the metastatic group, respectively. The Leu7+ CD16+ cells were not observed. The Leu7- CD16+ cells were observed in a part where metastases were focused. These results indicated that lymphocytes in regional lymphocyte of gastric cancer might hold the hidden anti-tumor effect, but did not display the full function without preoperative intratumor injection of BRM such as IL-2 or OK-432.
Assuntos
Linfonodos/patologia , Coloração e Rotulagem/métodos , Neoplasias Gástricas/imunologia , Linfócitos T/imunologia , Anticorpos Monoclonais , Imunofluorescência , Humanos , Células Matadoras Naturais/imunologia , Metástase Linfática , Ativação Linfocitária , Neoplasias Gástricas/patologia , Linfócitos T Auxiliares-Indutores/imunologiaRESUMO
An immunohistochemical study was performed on human lymphocytes in the tissue of gastric cancer, and also in the regional lymph nodes, by double fluorescent staining, using monoclonal antibodies. Leu3a+8+ cells (inducer T cells) which consist about 30 per cent of Leu 3a+ cells were seen in the tissue surrounding the gastric cancer. The other 70 per cent of Leu 3a+ cells were Leu3a+8- cells (helper T cells). In the lymph nodes they were noted in T cell areas in almost the same proportions, while in germinal centers, only Leu3a+8- cells were found. On the other hand, OKT8+Leu15- cells (cytotoxic T cells) were noted in a large number, while OKT8+Leu15+ cells (suppressor T cells) were few. Further, an increase of OKT8+Leu15- cells was seen around gastric cancer or metastatic cancer in lymph nodes. These immunohistochemical findings suggest that cytotoxic T cells are the main component in the tissue of gastric cancers and the regional lymph nodes. Increases in inducer T cells and helper T cells are probably required to induce cytotoxic T cells around the cancer tissue.
Assuntos
Linfonodos/patologia , Neoplasias Gástricas/patologia , Linfócitos T/patologia , Anticorpos Monoclonais , Imunofluorescência , Humanos , Estômago/patologia , Linfócitos T/classificaçãoRESUMO
To assess how endogenous prostaglandin (PG) in gastric mucosa acts against ulcer formation, we determined the mucosal prostacyclin (PGI2), PGE2, PGF2 alpha, and thromboxane A2(TXA2) concentrations before and after polypectomy in 6 patients in whom gastric ulcers were produced by electric burning resection of gastric polyps. These artificially induced ulcers all healed within short periods (25.7 +/- 7.4 days, mean +/- SE). Of the PGs assayed, the level of PGI2 was highest. The pG levels were increased at 4 and 7 days post-polypectomy; the most remarkable increase took place in the mucosa along the ulcer margin rather than the mucosa far from the ulcer site. We suggest that the observed increase in endogenous PGs represents a physiological response against polypectomy-induced ulcer formation.