基于FXR介导的胆汁酸重吸收探讨鲜地黄调节高脂高胆固醇饮食小鼠胆固醇代谢紊乱的作用机制  

Fresh Rehmanniae Radix regulates cholesterol metabolism disorder in mice fed with high-fat and high-cholesterol diet via FXR-mediated bile acid reabsorption

作　　者：孟鑫宇 陈燕 赵丽琴 刘庆普 靳永环 冯卫生[1,2] 郑晓珂 MENG Xin-yu;CHEN Yan;ZHAO Li-qin;LIU Qing-pu;JIN Yong-huan;FENG Wei-sheng;ZHENG Xiao-ke(School of Pharmacy,Henan University of Chinese Medicine,Zhengzhou 450046,China;Engineering and Technology Center for Chinese Medicine Development of Henan Province,Zhengzhou 450046,China;Henan Provincial Engineering Research Center for Quality Control and Evaluation of Traditional Chinese Medicine,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学药学院,河南郑州450046 [2]河南省中药开发工程技术研究中心,河南郑州450046 [3]河南省中药质量控制与评价工程技术研究中心,河南郑州450046

出　　处：《中国中药杂志》2025年第6期1670-1679,共10页China Journal of Chinese Materia Medica

基　　金：国家重点研发计划“中医药现代化”重点专项(2019YFC1708800);河南省高层次人才特殊支持计划项目(ZYQR201810080)。

摘　　要：明确鲜地黄水提物对高脂高胆固醇饮食诱导高胆固醇血症模型小鼠的改善作用,初步从胆汁酸重吸收角度探析其作用机制。将雄性C57BL/6小鼠随机分为正常组,模型组,鲜地黄低、高剂量(4、8 g·kg^(-1))组,阳性药辛伐他汀(0.05 g·kg^(-1))组。除正常组外,剩余组高脂高胆固醇饮食连续喂养6周建立高胆固醇血症模型小鼠,在第6周开始灌胃给药6周,期间继续高脂高胆固醇饮食喂养。检测小鼠血清总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、低密度动脉压脂蛋白胆固醇(low dendity lipoprotein-cholesterol,LDL-c)、高密度脂蛋白胆固醇(high dendity lipoprotein-cholesterol,HDL-c)、总胆汁酸(total bile acid,TBA)水平;检测肝脏TC、TG水平以及粪便TBA水平;采用苏木素-伊红(HE)、油红O染色以及透射电镜观察小鼠肝脏病理变化;随机选取正常组、模型组和鲜地黄高剂量组中每组3个肝组织样本进行高通量转录组测序,采用差异基因分析和KEGG通路富集分析筛选鲜地黄水提物治疗高胆固醇血症的基因和关键通路;采用RT-qPCR检测肝脏胆固醇7α-羟化酶(cholesterol 7α-hydroxylase,CYP7A1)、胆固醇27-羟化酶(cholesterol 27-hydroxylase,CYP27A1)的mRNA水平,Western blot检测肝脏CYP7A1、CYP27A1及回肠法尼醇X受体(farnesoid X receptor,FXR)、顶端钠依赖性胆盐转运体(apical sodium-dependent bile acid transporter,ASBT)、回肠胆汁酸结合蛋白(ileum bile acid binding protein,I-BABP)的蛋白表达。结果显示,鲜地黄水提物可明显降低小鼠血清和肝脏TC、TG水平以及血清LDL-c水平,升高血清HDL-c水平,并且能够增加粪便TBA水平,各组小鼠血清TBA水平无明显差异;HE、油红O染色以及透射电镜结果表明鲜地黄水提物可减轻肝脏脂质积累;进一步的机制探究发现,鲜地黄水提物能明显抑制FXR以及胆汁酸重吸收相关蛋白ASBT和I-BABP的表达水平,进而增强胆汁酸合成关键酶CYP7A1和CYP27This study aims to investigate the potential effect of the water extract of fresh Rehmanniae Radix on hypercholesterolemia in mice that was induced by a high-fat and high-cholesterol diet and explore its possible mechanism from bile acid reabsorption.Male C57BL/6 mice were randomly assigned into the following groups:control,model,low-and high-dose(4 and 8 g·kg^(-1),respectively)fresh Rehmanniae Radix,and positive drug(simvastatin,0.05 g·kg^(-1)).Other groups except the control group were fed with a high-fat and high-cholesterol diet for 6 consecutive weeks to induce hypercholesterolemia.From the 6th week,mice were administrated with corresponding drugs daily via gavage for additional 6 weeks,while continuing to be fed with a high-fat and high-cholesterol diet.Serum levels of total cholesterol(TC),triglycerides(TG),low density lipoprotein-cholesterol(LDL-c),high density lipoprotein-cholesterol(HDL-c),and total bile acid(TBA),as well as liver TC and TG levels and fecal TBA level,were determined by commercial assay kits.Hematoxylin-eosin(HE)staining,oil red O staining,and transmission electron microscopy were performed to observe the pathological changes in the liver.Three livers samples were randomly selected from each of the control,model,and high-dose fresh Rehmanniae Radix groups for high-throughput transcriptome sequencing.Differentially expressed genes were mined and KEGG pathway enrichment analysis was performed to predict the key pathways and target genes of the water extract of fresh Rehmanniae Radix in the treatment of hypercholesterolemia.RT-qPCR was employed to measure the mRNA levels of cholesterol 7α-hydroxylase(CYP7A1)and cholesterol 27α-hydroxylase(CYP27A1)in the liver.Western blot was employed to determine the protein levels of CYP7A1 and CYP27A1 in the liver as well as farnesoid X receptor(FXR),apical sodium-dependent bile acid transporter(ASBT),and ileum bile acid-binding protein(I-BABP)in the ileum.The results showed that the water extract of fresh Rehmanniae Radix significantly lowered the l

关 键 词：鲜地黄 高胆固醇血症 胆汁酸 胆固醇 

分 类 号：R285.5[医药卫生—中药学]