RESUMO
We examined the variation in plasma levels of endothelin (ET), calcitonin gene-related peptide (CGRP), nitric oxide (NO), and malondialdehyde (MDA), as well as superoxide dismutase (SOD) activity, in acute myocardial ischemia reperfusion injury in a rabbit model. Seventy rabbits were randomly assigned into 3 groups. Open-chest surgery (OCS) was performed for all rabbits. Group A (N = 20) received sham-surgery, group B (N = 25) was the reperfusion group, and group C (N = 25) was the infarction group. At 12 h after chest clo-sure, plasma ET levels in groups B and C were clearly increased, while CGRP levels were clearly decreased, particularly in group B. At 24 h after chest closure, ET levels were higher than before OCS, while there was no significant difference between groups B and C. ET in group B was decreased, while that in group C was increased at 12 h. No significant difference in CGRP was observed between 12 and 24 h after chest closure. NO levels in groups B and C at 12 h after chest closure were significantly decreased compared to those before OCS. NO levels in group B at 24, 48, and 72 h were significantly lower than those at 12 h, while those of group C were not significantly changed after 12 h. Dynamic monitoring and comparison of plasma levels of ET, CGRP, NO, and MDA as well as SOD activity revealed that appropriate intervention of these factors may reduce reperfusion injury.
Assuntos
Peptídeo Relacionado com Gene de Calcitonina/sangue , Endotelinas/sangue , Malondialdeído/sangue , Traumatismo por Reperfusão Miocárdica/sangue , Óxido Nítrico/sangue , Animais , Modelos Animais de Doenças , Humanos , Traumatismo por Reperfusão Miocárdica/patologia , Miocárdio/patologia , Coelhos , Superóxido Dismutase/sangueRESUMO
Intercellular adhesion molecule-1 (ICAM-1) is an important factor in the progression of inflammatory responses in vivo. To develop a new anti-inflammatory drug to block the biological activity of ICAM-1, we produced a monoclonal antibody (Ka=4.19×10−8 M) against human ICAM-1. The anti-ICAM-1 single-chain variable antibody fragment (scFv) was expressed at a high level as inclusion bodies in Escherichia coli. We refolded the scFv (Ka=2.35×10−7 M) by ion-exchange chromatography, dialysis, and dilution. The results showed that column chromatography refolding by high-performance Q Sepharose had remarkable advantages over conventional dilution and dialysis methods. Furthermore, the anti-ICAM-1 scFv yield of about 60 mg/L was higher with this method. The purity of the final product was greater than 90%, as shown by denaturing gel electrophoresis. Enzyme-linked immunosorbent assay, cell culture, and animal experiments were used to assess the immunological properties and biological activities of the renatured scFv.
Assuntos
Animais , Feminino , Humanos , Masculino , Camundongos , Expressão Gênica/fisiologia , Fragmentos de Imunoglobulinas/biossíntese , Molécula 1 de Adesão Intercelular/imunologia , Redobramento de Proteína , Renaturação Proteica , Anticorpos de Cadeia Única/biossíntese , Complexo Antígeno-Anticorpo , Anti-Inflamatórios/farmacologia , Anticorpos Monoclonais/biossíntese , Adesão Celular , Cromatografia , Diálise , Ensaio de Imunoadsorção Enzimática , Pavilhão Auricular/efeitos dos fármacos , Escherichia coli/genética , Vetores Genéticos , Fragmentos de Imunoglobulinas/farmacologia , Corpos de Inclusão/metabolismo , Molécula 1 de Adesão Intercelular/efeitos dos fármacos , Leucócitos Mononucleares/metabolismo , Plasmídeos , Engenharia de Proteínas/métodos , Anticorpos de Cadeia Única/farmacologia , Xilenos/farmacologiaRESUMO
Intercellular adhesion molecule-1 (ICAM-1) is an important factor in the progression of inflammatory responses in vivo. To develop a new anti-inflammatory drug to block the biological activity of ICAM-1, we produced a monoclonal antibody (Ka=4.19 × 10(-8) M) against human ICAM-1. The anti-ICAM-1 single-chain variable antibody fragment (scFv) was expressed at a high level as inclusion bodies in Escherichia coli. We refolded the scFv (Ka=2.35 × 10(-7) M) by ion-exchange chromatography, dialysis, and dilution. The results showed that column chromatography refolding by high-performance Q Sepharose had remarkable advantages over conventional dilution and dialysis methods. Furthermore, the anti-ICAM-1 scFv yield of about 60 mg/L was higher with this method. The purity of the final product was greater than 90%, as shown by denaturing gel electrophoresis. Enzyme-linked immunosorbent assay, cell culture, and animal experiments were used to assess the immunological properties and biological activities of the renatured scFv.