论文部分内容阅读
目的:建立胞内杀菌和体外中和内毒素实验模型,检测muBPI25目的蛋白对胞内寄生菌的抑杀作用和对内毒素的中和作用。方法:将pcDNA3.1(+)-muBPI36-259质粒导入RAW264.7细胞,用胞内寄生G+/G-菌感染上述细胞建立muBPI25目的蛋白胞内杀菌实验模型;将pSecTag2B-muBPI36-259与双荧光素酶报告基因质粒共转染RAW264.7细胞,建立体外检测muBPI25蛋白中和内毒素实验模型。结果:胞内杀菌实验模型证实muBPI25目的蛋白对G-伤寒杆菌具有抑杀作用;中和内毒素实验模型证实muBPI25目的蛋白对内毒素具有中和作用。结论:首次证实小鼠BPI N端功能片段,即muBPI25目的蛋白对G-菌具有抑杀作用,对其裂解产物内毒素具有中和作用。
OBJECTIVE: To establish a model of intracellular bactericidal and endogenous neutralization of endotoxin in vitro and to detect the inhibitory effect of the target protein of muBPI25 against intracellular parasites and the neutralization of endotoxin. Methods: The pcDNA3.1 (+) - muBPI36-259 plasmid was introduced into RAW264.7 cells and infected with the intracellular parasitic G + / G-bacteria to establish the intracellular bacteriostatic experiment of muBPI25 protein. The pSecTag2B-muBPI36-259 The luciferase reporter plasmid was co-transfected into RAW264.7 cells to establish an in vitro experimental model of muBPI25 protein neutralization and endotoxin. Results: The intracellular bactericidal experimental model confirmed that muBPI25 protein had an inhibitory effect on G-typhoid bacillus. Neutralization of endotoxin experimental model confirmed that muBPI25 protein had a neutralizing effect on endotoxin. CONCLUSION: It was first demonstrated that the N-terminal functional fragment of mouse BPI, namely the muBPI25 protein, has an inhibitory effect on G-bacteria and neutralizes endotoxin of its cleavage product.