上二黄丸通过调节胆汁酸代谢促进功能性便秘大鼠肠动力的配伍机制  

Compatibility Mechanism of Shangerhuang Wan in Enhancing Intestinal Motility in Rat Model of Functional Constipation by Regulating Bile Acid Metabolism

作　　者：曾耀莹 都广礼[1] ZENG Yaoying;DU Guangli(School of Traditional Chinese Medicine(TCM),Shanghai University of TCM,Shanghai 201203,China)

机构地区：[1]上海中医药大学中医学院,上海201203

出　　处：《中国实验方剂学杂志》2025年第6期1-8,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(81573862);2021年度高水平大学教学学术共同体项目(2021SHUTCMTD01)。

摘　　要：目的:基于胆汁酸代谢途径,探讨上二黄丸对洛哌丁胺诱导的功能性便秘模型大鼠的治疗作用及其配伍机制。方法:将60只大鼠随机分为6组,分别正常组、模型组、多潘立酮组(8 mg·kg^(-1))、升麻-柴胡组(1.33 g·kg^(-1))、黄芩-黄连组(1.20 g·kg^(-1))、上二黄丸全方组(3.33 g·kg^(-1)),每组10只。除正常组外其余大鼠每天2次皮下注射3 mg·kg^(-1)洛哌丁胺诱导7 d,即6 mg·kg^(-1)·d^(-1)建立功能性便秘模型,造模第3天开始相应地给药治疗,造模持续进行。采用炭末推进法检测大鼠小肠推进率,液相色谱-串联质谱法(LC-MS/MS)法检测粪便胆汁酸,微板法检测血清胆汁酸含量,酶联免疫吸附测定法(ELISA)检测结肠组织5-羟色胺(5-HT)和环磷酸腺苷(cAMP)水平,蛋白免疫印迹法(Western blot)检测回肠末端顶端Na+依赖性胆汁酸转运体(ASBT)和结肠跨膜G蛋白偶联受体5(TGR5)蛋白表达。结果:与正常组比较,模型组大鼠小肠推进率与粪便总胆汁酸显著降低(P<0.01),血清总胆汁酸升高,5-HT、cAMP水平显著降低(P<0.01),ASBT蛋白表达升高(P<0.01)、TGR5蛋白表达降低(P<0.05)。与模型组比较,多潘立酮组、全方组、芩连组小肠推进率升高(P<0.05,P<0.01),粪便总胆汁酸显著升高(P<0.01)、血清胆汁酸降低;与全方组比较,升柴组粪便总胆汁酸显著降低(P<0.01)、血清胆汁酸升高,芩连组粪便总胆汁酸显著升高(P<0.01)、血清胆汁酸降低。与模型组比较,多潘立酮组、全方组、芩连组ASBT蛋白表达明显降低(P<0.05,P<0.01);TGR5蛋白表达明显升高(P<0.05,P<0.01),升柴组ASBT蛋白表达差异无统计学意义,TGR5蛋白表达明显升高(P<0.05)。与模型组比较,多潘立酮组、全方组、芩连组5-HT水平和cAMP水平明显升高(P<0.05,P<0.01),升柴组5-HT水平差异无统计学意义,但cAMP水平显著升高(P<0.01)。结论:上二黄丸通过黄连-黄芩与升麻-柴胡配伍对胆汁酸代谢呈现正负相反性的调节,�Objective:To explore the mechanism of Shangerhuang Wan(SEHW)and its subdivisions in alleviating loperamide-induced functional constipation(FC)in rats by regulating bile acid metabolism.Methods:Sixty rats were randomly assigned into six groups(n=10):normal control,model,positive control(domperidone,8 mg·kg^(-1)),Cimicifugae Rhizoma-Bupleuri Radix(SC,1.33 g·kg^(-1)),Scutellariae Radix-Coptidis Rhizoma(QL,1.20 g·kg^(-1)),and SEHW(3.33 g·kg^(-1)).The remaining groups except the normal control group were subjected to subcutaneous injection with loperamide at 3 mg·kg^(-1)twice daily(total dose of 6 mg·kg^(-1)·d^(-1))for 7 days to induce a model of FC.Drug administration was initiated on day 3 of modeling,which continued throughout the modeling period.The small intestinal propulsion rate of each group was measured via the ink propulsion method.LC-MS/MS was employed to measure the fecal bile acid content in each group,and the serum bile acid level was measured by a microplate.The 5-hydroxytryptamine(5-HT)and cyclic adenosine monophosphate(cAMP)levels in the colon tissue of each group were measured by enzyme-linked immunosorbent assay(ELISA).Western blot was employed to determine the protein levels of apical sodium-dependent bile acid transporter(ASBT)in the ileum terminus and Takeda G protein-coupled receptor 5(TGR5)in the colon.Results:Compared with the normal control group,the model group exhibited decreases in the small intestinal propulsion rate and total fecal bile acid content(P<0.01),an elevation in the serum total bile acid level,lowered 5-HT and cAMP levels(P<0.01),up-regulation in the protein level of ASBT(P<0.01),and down-regulation in the protein level of TGR5(P<0.05).Compared with the model group,the positive control,SEHW,and QL groups showed increases in the small intestinal propulsion rate(P<0.05,P<0.01)and total fecal bile acid content(P<0.01)and a decline in the serum bile acid level.Compared with the SEHW group,the SC group had decreased total fecal bile acid content(P<0.01)and an elevated serum

关 键 词：上二黄丸 功能性便秘 胆汁酸代谢 肠动力 配伍 

分 类 号：R289[医药卫生—方剂学] R285[医药卫生—中药学] R259[医药卫生—中医学]