加减大柴胡汤对胆固醇结石湿热证小鼠的作用机制  

Mechanism of Modified Dachaihu Decoction on Cholesterol Gallstones Mice with Dampness-heat Syndrome

作　　者：闵莉[1] 王素英 章小燕 梁文娜[1] 沈建英[1] 周智慧 MIN Li;WANG Su-ying;ZHANG Xiao-yan;LIANG Wen-na;SHEN Jian-ying;ZHOU Zhi-hui(College of Traditional Chinese Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou,350122)

机构地区：[1]福建中医药大学中医学院,福州350122

出　　处：《中国中西医结合杂志》2023年第8期975-983,共9页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金项目(No.81603541);福建省自然科学基金项目(No.2021J01928,No.2017J01543)。

摘　　要：目的研究加减大柴胡汤对胆固醇结石(CS)湿热证小鼠胆固醇-胆汁酸代谢相关因子的影响,探讨其作用机制。方法将48只SPF级小鼠随机分为空白组、模型组、大柴胡汤组和熊去氧胆酸组,每组12只;空白组作为对照,其他3组采用“内湿+外湿+高胆固醇致石饲料”的模式造模12周,建立CS湿热证小鼠模型。造模成功后大柴胡汤组和熊去氧胆酸组分别给予加减大柴胡汤和熊去氧胆酸灌胃,模型组和空白组给予等量的生理盐水灌胃,连续干预4周。观察各组小鼠的结石及一般情况,进行旷场实验,NPP底物-AMP缓冲液法、GCNA底物法与钒盐酸氧化法分别检测血清碱性磷酸酶(ALP)、γ-谷氨酰基转移酶(GGT)、总胆红素(TBIL)含量,酶联免疫吸附测定法(ELISA)检测胆汁中总胆固醇(TC)、总胆汁酸(TBA)含量;实时荧光定量聚合酶链式反应(RT-PCR)及蛋白免疫印迹法(Western Blot)检测小鼠回肠组织中法尼酯衍生物X受体(FXR)、成纤维细胞生长因子15(FGF15)和肝脏组织中成纤维细胞生长因子受体4(FGFR4)、细胞外信号调节激酶(ERK)1/2、c-Jun氨基末端激酶(JNK)mRNA及蛋白表达水平。结果与空白组比较,模型组小鼠胆囊结石状况及湿热证候表现明显,胆汁TC增多、TBA减少(P<0.01),肝功能指标(ALP、GGT、TBIL)均显著升高(P<0.01),肝脏组织明显的脂肪样变性,回肠结构完整性被破坏,回肠FXR、FGF15与肝脏FGFR4、ERK1/2、JNK mRNA及蛋白表达均显著升高(P<0.05,P<0.01)。与模型组比较,大柴胡汤组小鼠胆囊结石状况、肝脏和回肠组织病理形态以及湿热证候表现改善明显;大柴胡汤组和熊去氧胆酸组小鼠胆汁内TC含量均显著降低(P<0.01)、TBA含量均显著增高(P<0.01),血清ALP、GGT、TBIL含量均显著降低(P<0.01)。与熊去氧胆酸组比较,大柴胡汤组血清肝功能指标和胆汁成分改善更显著(P<0.01)。与模型组比较,两用药组小鼠FXR、FGF15、FGFR4、ERK1/2、JNK mRNA和�Objective To study the effects of Modified Dachaihu Decoction(MDD)on related factors of cholesterol gallstones(CS)mice with dampness heat syndrome,and to explore its mechanism of action.Methods Forty-eight SPF grade mice were randomly divided into blank group,model group,Dachaihu Decoction group,and ursodeoxycholic acid group,12 mice in each group.The blank group was set as the control group,and the other 3 groups were modeled with the model of"internal dampness+external dampness+high cholesterol stone-induced forage"for 12 successive weeks to establish CS dampness-heat syndrome model.After successful modeling mice in Dachaihu Decoction group and ursodeoxycholic acid group were respectively administered with MDD and ursodeoxycholic acid by gastrogavage for 4 successive weeks.Mice in model group and blank group were administered with the same amount of normal saline by gastrogavage for 4 successive weeks.The calculi and general conditions of mice were observed,and the open field test was conducted.The contents of alkaline phosphatase(ALP),γ-glutamyltransferase(GGT),and total bilirubin(TBIL)in serum were detected by NPP substrate-AMP buffer method,GCNA substrate method and vanadium hydrochloric acid oxidation method,respectively.The contents of total cholesterol(TC)and total bile acid(TBA)in bile were detected by enzyme-related immunosorbent assay(ELISA).Real-time fluorescence quantitative polymerase chain reaction(RT-PCR)and Western Blot were used to detect the mRNA expression levels and protein expression levels of FXR and fibroblast growth factor(FGF)15 in mouse ileum and fibroblast growth factor receptor 4(FGFR4),extracellular regulated protein kinase 1/2(ERK1/2)and C-Jun N-terminal protein kinase(JNK)in liver.Results Compared with blank group,gallstone conditions and manifestations of dampness-heat syndrome were obvious in model group,bile TC increased,TBA decreased(P<0.01),liver function indices(ALP,GGT,TBIL)increased significantly(P<0.01),fatty degeneration of liver tissue was obviously seen,ilum structural

关 键 词：胆囊胆固醇结石 湿热证 大柴胡汤 胆固醇-胆汁酸代谢 中药复方 

分 类 号：R285.5[医药卫生—中药学]