豚鼠胆囊胆固醇结石形成过程中肠道传输功能下降的细胞及分子机制探讨  

Cellular and molecular mechanism of slow intestinal transit during formation of cholesterol gallstones in guinea pig model

作　　者：范莹[1] 吴硕东[1] 殷振华[1] 付倍蓓[2] 

机构地区：[1]中国医科大学附属盛京医院普外二科,辽宁沈阳110004 [2]中国医科大学附属盛京医院小儿血液内科,辽宁沈阳110004

出　　处：《生物医学工程与临床》2013年第1期67-71,共5页Biomedical Engineering and Clinical Medicine

基　　金：国家自然科学基金资助项目(81000183);沈阳市科学技术计划资助项目(F11-264-1-24)

摘　　要：目的探讨豚鼠胆囊胆固醇结石形成过程中肠道传输功能下降的细胞和分子机制及其与胆石形成的关系。方法健康雄性豚鼠40只,4周龄,体质量120～125g。将其随机分为实验组与对照组,每组20只。实验组给予致石饲料(胆固醇含量2%),对照组给予正常颗粒饲料。8周造模结束后,用逆转录聚合酶链反应(RT-PCR)检测小肠组织中c-kit及scf的mRNA的表达情况,利用末端回肠全层铺片免疫荧光化学染色及激光共聚焦显微镜观察各组Cajal样间质细胞(ICCs)数量的变化。结果 RT-PCR结果显示,与对照组相比,实验组豚鼠小肠c-kit(0.316±0.056vs0.912±0.103;t=6.582,P<0.01)和scf(0.499±0.012 vs 0.899±0.124;t=6.163,P<0.01)的mRNA水平的表达量下降;对照组豚鼠回肠ICCs平均阳性面积为(56.24±2.68)%,实验组为(22.26±1.14)%,较对照组明显降低(t=15.256,P<0.01)。结论饮食诱导的豚鼠胆囊胆固醇结石形成过程中,小肠c-kit和scf基因mRNA表达水平下降,ICCs数量明显减少。c-kit/scf通路抑制可能参与胆囊胆固醇结石的形成过程中肠道传输功能下降的发生。Objective To investigate the cellular and molecular mechanism of slow intestinal transit during the formation of cholesterol gallstones in guinea pig model,and study its relationship with gallstone formation.Methods Forty healthy male guinea pigs of 4-week old and 120-150 g body weight were randomly divided into 2 groups,namely experimental group and control group（n = 20）.The experimental group was fed high cholesterol diet（cholesterol 2 %）,and control group fed normal diet.The guinea pigs were sacrificed after 8 weeks.Reverse transcription polymerase chain reaction（RT-PCR） was used to determine the expression of c-kit and scf mRNA.Change in the numbers of interstitial Cajal cells（ICCs） in terminal ileum was observed by immunohistochemical method.Results Compared with the control group,the mRNA expression of c-kit（0.316 ± 0.056 vs 0.912 ± 0.103;t = 6.582,P 0.01） and scf（0.499 ± 0.012 vs 0.899 ± 0.124;t = 6.163,P 0.01） was decreased significantly in the experimental group.Compared with the control group,the average positive area of ICCs was decreased obviously in the experimental group（22.26 % ± 1.14 % vs 56.24 % ± 2.68 %;t = 15.256,P 0.01）.Conclusion It is demonstrated that the expression of c-kit/scf mRNA and numbers of ICCs in the small intestine are decreased during the formation of cholesterol gallstones in guinea pigs fed on high cholesterol diet.Inhibition of c-kit/scf pathway may play a role in the pathogenesis of slow intestinal transit during the formation of cholesterol gallstones.

关 键 词：胆囊胆固醇结石 肠道传输 CAJAL间质细胞 C-KIT 干细胞因子 

分 类 号：R575.62[医药卫生—消化系统]