骨桥蛋白在不同胆汁体系中的成核作用研究
The role of osteopontin in nucleation in different bile systems
陈进宏 1杨林 1蔡端 2王丽颖 3查锡良3
作者信息
- 1. 200040上海,复旦大学附属华山医院普外科;西安交通大学医学院第一附属医院外科
- 2. 200040上海,复旦大学附属华山医院普外科
- 3. 复旦大学医学院生化系
- 折叠
摘要
目的 探讨骨桥蛋白( Osteopontin,OPN)在不同胆汁体系中的成核作用,研究其与胆结石形成的关系.方法 应用成核时间实验检测OPN在模拟胆汁及人胆汁中的成核时间.胆固醇结晶生长实验检测OPN对模拟胆汁胆固醇结晶生长的影响,电镜观察OPN对模拟胆汁及人胆汁中胆汁泡的影响,并检测胆结石患者胆汁和正常胆汁中OPN和钙离子的浓度.结果 OPN在模拟胆汁及人胆汁中均可延长胆固醇成核时间,其作用具有剂量依赖性并和钙离子相关(P＜0.05).50 μg/ml和100 μg/ml的OPN分别使结石胆汁的成核时间延长1.50倍和1.93倍(P＜0.01)；使正常胆汁的成核时间延长1.17倍和1.33倍(P＜0.01)；使模拟胆汁的成核时间延长1.29倍和1.48倍(P＜0.01).在模拟胆汁中,OPN可抑制胆固醇结晶生长,延缓胆固醇结晶出现,使胆固醇成核时间指数和胆固醇结晶生长指数延长,并可抑制由钙离子诱导的促成核效应.电镜观察发现OPN可使胆固醇磷脂泡更加稳定,抑制泡的动态变化及胆固醇析出.结石胆汁中OPN[(0.53±0.08)mg/ml比(0.65±0.14) mg/ml,P＜0.05]和钙离子浓度均低与对照组[(0.71±0.17) mmol/L比(0.84±0.08) mmol/L,P＜0.05].结论 OPN在不同胆汁体系中均可抑制胆固醇成核,从而抑制胆固醇结石形成.
Abstract
Objective To investigate the role of osteopontin (OPN) in the pathogenesis ot cholesterol gallstone formation in bile.Methods The nucleation time of OPN in model bile and human gallbladder bile was studied by the nucleation time assay,the effect of OPN on cholesterol crystal growth in model bile was examined by the cholesterol crystal growth assay.The effect of OPN on vesicle was detected by the transmission electron microscopy in model bile and gallbladder bile; then the content of OPN and calcium were detected via the commercial kits in human bile.Results Osteopontin prolonged nucleation time in a dose dependent manner in model bile and human bile,and this effect was correlated with calcium.Compared with control group,the nucleation times were prolonged by 1.50and 1.93 times in lithogenic bile at the concentration of osteopontin 50 μg/ml and 100 μg/ml (P＜0.01),respectively. Nucleation time were prolonged by 1.17 and 1.33 times in normal bile (P＜0.01) and by 1.29 and 1.48 times in model bile (P＜0.01),respectively.The rate of cholesterol crystals growth was not influenced by calcium ions,but inhibited by osteopontin in a dose dependent manner in the model bile.Furthermore,the formation,aggregation and fusion of vesicles were delayed by osteopontin in bile samples as indicated by the transmission electron microscopy.The concentration of osteopontin [(0.53± 0.08) mg/ml vs. (0.65 ± 0.14) mg/ml,P＜0.05] and the calcium ions [ (0.71 ± 0.17) mmol/L vs. ( 0.84 ± 0.08 ) mmol/L,P ＜ 0.05 ] were lower in lithogenic bile than in control.Conclusions Osteopontin can inhibit the cholesterol gallstone formation in model and human gallbladder bile as the anti nucleating factor.
关键词
胆结石/骨桥蛋白/成核作用/胆汁Key words
Cholelithiasis/Osteopontin/Nucleation role/Bile引用本文复制引用
出版年
2012
NETL
NSTL
维普
万方数据