猪中性粒细胞抑菌蛋白的分离及对细菌影响

Purification and identification of neutrophil defensins in porcine blood

目的 提取、分离、纯化、高效高活性杀烧伤创面病原菌的猪血中性粒细胞防御素。方法 用 5 %乙酸提取猪血中性粒细胞溶酶体可溶性成分 ,经制备性聚丙烯酰胺凝胶电泳纯化猪中性粒细胞防御素 ,用SDS PAGE检测防御素的分子质量 ,以体外杀烧伤创面白色念珠菌试验、体外抑制烧伤创面金黄色葡萄球菌和绿脓杆菌的生长作用检测其活性。结果 可从提取物中纯化出 7种多肽 (ND1、ND2、ND3、ND4、ND5、ND6、ND7) ,它们的分子质量在 2 80 0～ 680 0之间。体外杀白色念珠菌试验和体外抑制金黄色葡萄球菌及绿脓杆菌的生长作用证实 ,ND1、ND2有高效杀菌活性 ,ND3也有较强的杀菌活性。结论 通过猪血中性粒细胞的分离、纯化 。

Objective To extract, isolate and purify highly effective neutrophil defensins(ND) which can kill bacteria in burn wound from porcine blood. Methods The soluble part of the neutrophil lysosome in porcine blood was extracted by using 5% ethanoic acid. Preparative SDS PAGE was employed to purify neutrophil defensins. The molecular weight (MW) of defensins was detected by SDS PAGE. The activity of defensins was determined by tests of killing of candida albicans from burn wound and inhibition of staphylococcus aeurus and pseudomonas aeruginosa growth in vitro . Results Seven kinds of polypeptides (ND1～ND7 with MW at 2 800～6 800) could be purified from the extracted substance. The above in vitro killing and inhibition tests confirmed that ND1, ND2 and ND3 (less strong) exhibited highly effective bactericidal activity. Conclusion Defensins with highly effective bactericidal activity can be obtained by the isolation and purification of neutrophils in porcine blood.