摘要
目的探讨运甲状腺素蛋白(transthyretin,TTR)基因及其蛋白在胆固醇结石患者胆囊组织中的表达情况,以及其在胆固醇结石形成中的促成核作用。方法收集笔者所在的两所医院施行择期胆囊切除术的25例胆固醇结石患者和9例肝移植供体的正常胆囊组织,采用逆转录-聚合酶链反应(RT-PCR)技术及Western blot法检测2组患者胆囊组织中TTR基因及其蛋白的表达。构建Small和综合模拟胆汁体系,分别加入TTR和白蛋白(ALB),再通过Holan成核时间法和Holzbach法分别检测成核时间和成核活性。结果胆固醇结石组TTR mRNA及其蛋白的表达水平分别为1.51±0.78和3.95±0.09,均高于正常对照组的0.85±0.63和1.53±0.08(P<0.05)。在Small模拟胆汁体系中,观察至21 d时,TTR组的成核时间为(14.5±1.3)d,ALB组为(18.0±0.8)d,TTR的成核活性为0.81;在综合模拟胆汁体系中,TTR组的成核时间为(13.5±0.6)d,ALB组为(18.5±1.3)d,TTR的成核活性为0.73。TTR组的成核时间均较短(P<0.01)。结论 TTR在胆固醇结石患者胆囊组织中的表达上调,并在体外模拟胆汁体系中表现出促成核活性,提示其参与了胆固醇结石的发生和发展过程。
Objective To explore the expressions of transthyretin (TTR) mRNA and its protein in tissues of human gallbladder with cholesterol gallstones, and to explore its role in the formation of cholesterol gallstones. Methods Gallbladder were got from cases of cholesterol gallstones (cholesterol gallstones group, n=25) and cases underwent liver transplantation with normal gallbladder (normal control group, n=9) respectively, who were treated in Ren ji Hospital and Huashan Hospital. Real time PCR (RT-PCR) and Western blot method were used to determine the expressions of TTR mRNA and its protein respectively. In addition, 2 kinds of artificial model bile system were established to test nucleation time (NT) and nucleation activity, which added TTR and albumin (ALB). Results The expression levels of TTR mRNA and protein in cholesterol gallstones group were 1.51±0.78 and 3.95±0.09 respectively, which were both higher than those of normal control group (P〈0.05). The NT were (14.5±1.3) d and (18.0±0.8) d in TTR group and ALB group in small model bile system (P〈0.01), which was similar with comprehensive model bile system [(13.5±0.6)d vs. (18.5±1.3)d] . The nucleation activity were 0.81 and 0.73 in small model bile system and comprehensive model bile system respectively. Conclusions Expression of TTR up-regulates in human gallbladder tissues of patients with cholesterol gallstones, and TTR plays role of nucleation in model bile system, which is related to the formation of cholesterol gallstones.
出处
《中国普外基础与临床杂志》
CAS
2014年第6期716-720,共5页
Chinese Journal of Bases and Clinics In General Surgery