大黄灵仙方对胆固醇结石豚鼠模型胆囊Cajal间质细胞中scf/c-kit信号通路的影响  

Effect of Dahuang Lingxian prescription on the scf/c-kit signaling pathway in gallbladder interstitial cells of Cajal in a guinea pig model of cholesterol gallstone

作　　者：肖丽君 甘苡榕 刘春丽 李承积 杨文 庞浇安 滕金豪 俞渊[2] XIAO Lijun;GAN Yirong;LIU Chunli;LI Chengji;YANG Wen;PANG Jiaoan;TENG Jinhao;YU Yuan(Graduate School of Guangxi University of Chinese Medicine,Nanning 530001,China;Department of Hepatobiliary Surgery,The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,China)

机构地区：[1]广西中医药大学研究生学院,南宁530001 [2]广西中医药大学第一附属医院肝胆外科,南宁530023

出　　处：《临床肝胆病杂志》2023年第2期376-382,共7页Journal of Clinical Hepatology

基　　金：广西自然科学基金项目(2020GXNSFAA238012);2021年“岐黄工程”高层次人才团队培育项目(2021006);2020年广西中医药大学第一附属医院院级博士启动基金项目(2020BS004);广西中医药大学2021年研究生教育创新计划项目(YCXJ2021068)。

摘　　要：目的 观察在大黄灵仙方的调控下干细胞生长因子(scf)、酪氨酸激酶受体(c-kit)的表达水平,探讨其影响胆囊动力学改变的可能机制,为大黄灵仙方预防胆石症的发生及复发提供理论依据。方法 将45只SPF级健康雄性豚鼠随机分为正常组、模型组、中药组,其中正常组予正常饲料喂养,模型组、中药组予高脂致石饲料喂养,喂养8周后分别从各组中随机抽取5只豚鼠,肉眼观察下结石形成超过4只,则判断为模型建立成功。造模成功后中药组予大黄灵仙方灌胃,模型组予等体积生理盐水灌胃,连续灌胃给药8周后取豚鼠胆囊组织,HE染色观察观察胆囊组织的病理学改变,Western blot测定胆囊组织中TNFα的表达水平,免疫组化测定胆囊平滑肌组织中scf、c-kit的蛋白表达水平。符合正态分布的计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD多重比较法。结果 HE染色显示模型组胆囊组织炎症明显,中药组胆囊组织内炎症程度较模型组明显减轻。Western blot结果表明在模型组中,胆囊组织中TNFα的表达水平最高,中药组次之,正常组最低(P值均<0.05);免疫组化结果表明豚鼠胆囊平滑肌组织中scf、c-kit蛋白在正常组、中药组中的表达水平显著高于模型组(P值均<0.05)。结论 大黄灵仙方能增强胆囊动力功能,其作用机制可能与上调胆囊Cajal间质细胞中的scf、c-kit信号通路有关。Objective To investigate the expression levels of scf and c-kit under the regulation of Dahuang Lingxian prescription and the possible mechanism of its effect on gallbladder dynamics,and to provide a theoretical basis for Dahuang Lingxian prescription in preventing the development and recurrence of cholesterol gallstone.Methods A total of 45 specific pathogen-free healthy male guinea pigs were randomly divided into normal group,model group,and traditional Chinese medicine(TCM) group.The guinea pigs in the normal group were fed with normal diet,and those in the model group and the TCM group were fed with high-fat lithogenic diet.After 8 weeks of feeding,5 guinea pigs were randomly selected from each group,and successful modeling was determined if gallstone was observed with the naked eye in more than 4 guinea pigs.After successful modeling,the guinea pigs in the TCM group were given Dahuang Lingxian prescription by gavage,and those in the model group were given an equal volume of normal saline by gavage.After 8 consecutive weeks of administration by gavage,gallbladder tissue samples were collected,and HE staining was used to observe the pathological changes of gallbladder tissue;Western blot was used to measure the expression level of tumor necrosis factor-α(TNF-α) in gallbladder tissue;immunohistochemistry was used to measure the protein expression levels of scf and c-kit in gallbladder smooth muscle tissue.A one-way analysis of variance was used for comparison of normally distributed continuous data between multiple groups,and the least significant difference multiple comparison method was used for further comparison between two groups.Results HE staining showed marked inflammation of gallbladder tissue in the model group,and compared with the model group,the TCM group had a significantly lower degree of inflammation.Western blot showed that the model group had the highest expression level of TNF-α in gallbladder tissue,followed by the TCM group and the normal group(P<0.05);immunohistochemistry showed that co

关 键 词：大黄灵仙方 胆石 Cajal小肠细胞 信号传导 豚鼠 

分 类 号：R285.5[医药卫生—中药学]