胆石症病人和正常人胆汁泡蛋白成核活性及糖链结构的比较

Study of pro-nucleating activity and structure of sugar chain on 33. 5 kDa vesicular protein in bile from normal persons and cholesterol gallstone patients

目的比较胆石症病人和正常人胆汁泡蛋白促成核活性，并分析其糖链结构特点。方法进行胆固醇结晶生长试验，检测胆石症病人和正常人相对分子质量为33500的泡蛋白成核时间，并计算其促成核活性。采用HRP-凝集素探针介导的Western Blot印迹法分析不同来源泡蛋白糖链的结构特点。结果胆石症病人来源的33500泡蛋白具有明显的促成核活性，其平均成核时间为（3．75±0．66）d，成核活性为0．303±0．075；而正常人来源者则仅具较弱的促成核活性，平均成核时间为（8．38±0．86）d，成核活性为0．677±0．086，两者比较，差异显著（P〈0．05）。胆石症病人来源的33500泡蛋白DSA（＋）、ConA（-），属于复杂型多天线聚糖；而正常人来源者为ConA（＋）、DSA（-）的二天线糖链。结论胆石症病人来源的33500泡蛋白较正常人来源者具有更强的促成核活性，两者糖链结构特点亦有明显不同，提示泡蛋白促成核活性的改变可能由于其糖链结构特点的变化引起。

Objective To compare the pro-nucleating activities and analyze the similarity and difference of the structure of the sugar chain on 33.5 kilodalton （kDa） vesicular protein extracted from fresh native bile in different persons. Methods The 33.5 kDa vesicular protein was compared for pro moting activities with a sensitive cholesterol crystal growth assay and detected by Western blot meth od. Results The average nucleation time （NT） tested by Holan technique of the 33.5 kDa vesicular protein from cholesterol gallstone patients was 3. 75±0. 66 days, the pro nucleating activity was 0. 303. But the 33.5 kDa vesicular protein from normal person was weakly to nucleate, the average nucleation time （NT） was 8.38±0.86 days, and the pro-nucleating activity was only 0. 677. The Analytical results manifested that the number of the antennas in sugar chain of the 33.5 kDa vesicular protein was the main difference between cholesterol gallstone patients and healthy persons. Conclusion The pro-nucleating activity alteration of the vesicular protein may correspond closely with the increase of the number of the antennas in sugar chain on the 33.5 kDa vesicular protein when the normal bile transforms to pathological bile.