建立SP-A检测法测定体外循环前后猪SP-A含量变化

Detection of surfactant protein A in bronchoalveolar lavage fluid of pigs before and after CPB

目的 建立肺表面活性蛋白A(SP -A)的整套检测方法 ,并以之测定体外循环前后幼猪的支气管肺泡灌洗液 (BALF)中SP -A含量。方法 灌洗新鲜猪肺取得BALF ,经过抽提、透析、离心、亲和柱层析 ,获得SP -A。以之免疫大白兔 ,得到兔抗猪SP -A抗体。使用该抗体 ,以蛋白免疫印迹反应法检测 9头幼猪在体外循环 (CPB)前、CPB 1 2 0min、CPB后 1 80min采集的BALF中SP -A含量。结果 制备出猪SP -A和兔抗猪SP -A抗体。成功测定幼猪体外循环前后BALF中SP -A含量。结论 蛋白免疫印迹反应法测定SP -A方法的建立 。

Objective To establish methods for detection of surfactant protein A (SP-A) in bronchoalveolar lavage fluid (BALF) of young pigs before and after cardiovascular bypass (CPB). Methods Procine lungs were lavaged and BALF was collected. Pure SP-A was isolated from BALF by means of extration, dialysis, centrifugation and affinity column chromagraphy. Antibody against procine SP-A was raised by immunation of rabbits with SP-A. Such antibody was used in western-blotting, to detect SP-A in BALF samples of nine young pigs before CPB, CPB for 120 minutes and 180 minutes after CPB. Results Pure procine SP-A and antibody against SP-A were produced. SP-A in BALF of young pigs before and after CPB was successfully detected. Conclusion Detection of SP-A by Western-blotting facilitates studies on the effects of CPB on pulmonary surfactant.