【摘要】 目的 纯化并比较各种成核效应蛋白的活性,筛选最具病理学意义的促成核因子。方法 通过ConA-Sepharose 4B亲和层析和Sephadex G-200、Sephadex G-100分子筛凝胶过滤以及超速离心法从胆固醇性结石病人胆汁中分离提纯70 kDa、200 kDa促成核蛋白以及33.5 kDa泡蛋白,在模拟胆汁体系中观察成核因子促进胆固醇结晶生长、析出过程,比较其成核活性。结果 四种候选的成核效应蛋白中,33.5 kDa泡蛋白是活性最强的促成核因子,成核活性强弱依次为33.5 kDa泡蛋白>200 kDa糖蛋白≈α1-酸性糖蛋白>70 kDa糖蛋白,当浓度为100／μg／ml时,33.5 kDa泡蛋白成核活性指数Ig、Ic、It分别为1.52、1.63、0.57,并随着蛋白浓度的增加,促成核活性亦随之增强(F=678.18,P<0.05),且呈剂量依赖效应。结论 胆汁中存在多种促成核作用的糖蛋白,其中33.5 kDa泡蛋白可能与胆石形成关系最为密切。更多还原

【Abstract】 <abstract>jective To immunopurify and examine the comparative activities of the candidate pro-nucleating proteins. Methods The 70 kDa and 200 kDa glycoproteins were isolated by immu-naffinity chromatography using ConA-Sepharose 4B, Sephadex G-200 and Sephadex G-100, and the 33. 5 kDa vesicular protein was purified by ultracentrifugation from cholesterol gallstone bile. Comparative effect of adding crystallization-promoters was detected by cholesterol crystal growth. Results A-mongst the 4 glycoproteins, the 33. 5 vesicular protein showed the strongest promoting effects. The potency of these 4 proteins was in the following order: 33. 5 kDa vesicular protein > 200 kDa glyco-protein ≈α1-acid glycoprotein > 70 kDa glycoprotein. The activity indexes Ig, Ic and It of the 33. 5 kDa vesicular protein was 1. 52, 1.63 and 0. 57 respectively, at the concentration of 100 μg/mlo in model bile, and the pro-nucleating activity was enhanced along with increase in concentration of the protein. Conclusions Several glycoproteins exist in the bile may account for the gallstone formation and the 33. 5 kDa vesicular protein plays an important role in cholesterol crystallization.更多还原