33.5kDa胆汁泡蛋白促成核活性及其对泡形态学影响的研究

The potent cholesterol nucleation promoting activity and morphometric effect on the model biliary vesicles of 33.5kDa vesicular protein

目的探讨３３．５ｋＤａ泡蛋白在胆固醇成核过程中的作用。方法利用超速离心技术结合ＰＡＧＥ电泳及区带定位法制备３３．５ｋＤａ泡蛋白；并将其加入Ｓｍａｌ模拟胆汁及综合模拟胆汁作为试验组，相应地以人体白蛋白为对照组，用偏光显微镜观察胆固醇成核时间，在透射电镜下观察模拟胆汁泡的大小、形态及数量。结果Ｓｍａｌ试验组成核时间为３．８３±０．３１天，对照组为１２．３３±０．５６天（Ｐ＜０．００１）；综合试验组平均成核时间为３．１７±０．１７天，而对照组长达１０．３３±０．２１天（Ｐ＜０．００１）。随着时间的推移，Ｓｍａｌ试验组及综合试验组泡逐渐增大、聚集、融合，最终形成巨大复层泡。两组均较相应的对照组变化显著（Ｐ＜０．０５）。结论３３．５ｋＤａ泡蛋白具有很强的促进泡形成、聚集、融合及胆固醇单水结晶析出的作用（成核活性约为０．３１０）。

Objective:To examine the role of 33.5kDa vesicular protein in cholesterol nucleation process.Methods:Compare albumin with the vesicular protein mixed with Small artificial model bile or synthetic artificial model bile respectively.All bile samples were examined by polarizing microscopy to detect nucleation time,and to observe size,figure and quantity of model biliary vesicles under transmission electron microscopy.Results:33.5kDa vesicular protein shortened the nucleation time of cholesterol monohydrate crystals when added to Small artificial mode bile or synthetic artificial model bile,while albumin did not.(3.83±0.31vs 12.33±0.56 days,3.17±0.17vs 10.33±0.21 days respectively,P<0.001).Furthermore,a strikingly rapid de novo formation of unilamellar vesicles was found soon followed by massivevesicular aggregation,culminatingly rapid denovo formation of unilamellar vesicles was found,Soon followed by massive vesicular aggregation,culmination crystal formation.The potent cholesterol nucleation promoting actovities of 33.5kDa vesicular proteins were almost indiscriminate in the two different artificial modle biles.Conclusions:These data suggest that 33.5kDa vesicular proteins play an important role in vesicle formation,aggregation and fusion and accelerate the rate of formation of solid cholesterol crystals and thus may help to promote spontaneous cholesterol gallstone formation in humans.