达立通颗粒主要活性成分在正常及功能性消化不良模型大鼠胃肠道组织中差异分布研究  

Differential distribution of main active components in Dalitong Granules in gastrointestinal tissues of normal and functional dyspepsia model rats

作　　者：苏艳 康安[1] 赵岩 王晓霞 邝振英 盛先杰 王广基[3] 阿基业[3] SU Yan;KANG An;ZHAO Yan;WANG Xiaoxia;KUANG Zhenying;SHENG Xianjie;WANG Guangji;AA Jiye(School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023,China;Nanchang Helioeast Pharmaceutical Co.,Ltd.,Nanchang 330006,China;Jiangsu Key Laboratory of Drug Metabolism and Pharmacokinetics,China Pharmaceutical University,Nanjing 210009,China)

机构地区：[1]南京中医药大学药学院,江苏南京210023 [2]南昌弘益药业有限公司,江西南昌330096 [3]中国药科大学江苏省药物代谢动力学重点实验室,江苏南京210009

出　　处：《药物评价研究》2024年第5期1017-1029,共13页Drug Evaluation Research

基　　金：中国医学科学院医学与健康科技创新工程项目(CIFMS:2021-I2M-5-011);江苏省前沿引领技术基础研究专项拟立项目(BK20192005)。

摘　　要：目的研究达立通颗粒主要活性成分在正常和功能性消化不良(FD)模型大鼠胃肠道组织中的分布差异。方法采用夹尾刺激结合不规律饮食双因素干预方法建立FD模型。12只大鼠(对照、模型各6只)用于FD模型验证,进行胃、小肠排空实验,HE染色后进行胃肠组织病理学观察;36只大鼠(对照、模型各18只)用于胃肠道组织差异分布研究,禁食不禁水18 h,按22.4 g‧kg^(−1)剂量(8倍临床等效剂量)ig给予达立通颗粒1次,于给药后1、3、6 h收集各组大鼠的胃、十二指肠、空肠、回肠和结肠。应用超高效液相色谱-串联四级杆/线性离子阱质谱(UPLC-QTRAP-MS/MS)法检测各组织牡荆素-4''-O-葡糖苷、原阿片碱、黄连碱、小檗碱、甜橙黄酮、川陈皮素、桔皮素、去甲基川陈皮素、木香烃内酯、去氢木香内酯、原儿茶酸、伞形花内酯、木犀草苷、橙皮苷、木犀草素、柚皮素、橙皮素、白杨素含量,采用ACQUITY HPLCTM HSS T3(50 mm×2.1 mm,1.8μm)色谱柱,柱温为40℃,0.1%甲酸水-乙腈为流动相,梯度洗脱,体积流量为0.3 mL‧min^(−1),进样量为2μL。结果与对照组比较,模型组大鼠体质量显著降低(P<0.001),胃、小肠排空率显著降低(P<0.001),十二指肠肠黏膜的紧密连接被破坏,但未出现胃肠组织的器质性病变,符合FD的病理特征。建立的UPLC-QTRAP-MS/MS方法中,各化合物线性关系良好,R^(2)≥0.9900。对建立的检测方法进行专属性、精密度与准确度、提取回收率、稳定性和基质效应等方面的考察,均符合要求。各成分在对照和模型组大鼠胃肠道组织中浓度差异明显,且在各组织中达到最高浓度的时间点差异较大,模型组出现达峰时间延后的现象;与对照组比较,模型组大鼠胃组织中化合物的暴露水平降低,可能与FD引起胃肠道蠕动减慢有关。橙皮苷、橙皮素、柚皮素、原儿茶酸、去氢木香内酯、川陈皮素、去甲基川陈皮素、小�Objective The aim of this study was to explore the gastrointestinal tissue distribution differences of the main active compounds from Dalitong granules in normal and functional dyspepsia model rats.Method The rat model of FD was established by tail clipping stimulation combined with irregular diet(alternate day fasting).12 rats(half of them as controls and half as models)were used to validate the FD model,and gastric and small intestinal emptying experiments were conducted.After HE staining,pathological observations were made on the gastrointestinal tissues.36 rats(half as controls and half as models)were used for the study of differential distribution of gastrointestinal tissues,and they were fasted but not water-restricted for 18 hours.They were given Daliting Granules at a dose of 22.4 g·kg^(−1)(8 times the clinical equivalent dose)once,and the gastric,duodenum,jejunum,ileum,and colon were collected from each group of rats at 1,3,and 6 hours after the administration of the drug.The contents of vitexin-4''-O-glucoside,protopine,coptisine,berberine,sinensetin,nobiletin,tangeretin,5-O-demethylnobiletin,costunolide,dehydrocostuslactone,protocatechuic acid,umbelliferone,cynaroside,hesperidin,luteolin,naringin,hesperetin,and chrysin were detected by UPLC-QTRAP-MS/MS method in the gastric,duodenum,jejunum,ileum,and colon of each group of rats.UPLCQTRAP-MS/MS was used.Column was an ACQUITY HPLCTM HSS T3(50 mm×2.1 mm,1.8μm)and set at 40℃.Gradient elution was performed using 0.1%formic acid water and acetonitrile as the mobile phase,the flow rate was 0.3 mL‧min^(-1) and the injection volume was 2μL.Results Compared with the control group,the body mass of rats in the model group was significantly decreased(P<0.001),the emptying rate of the stomach and small intestine was significantly decreased(P<0.001),the close connection of the duodenal mucosa was destroyed,but no organic lesions of the gastrointestinal tissue were found,which was consistent with the pathological characteristics of FD.For the established UPL

关 键 词：达立通颗粒 功能性消化不良 UPLC-QTRAP-MS/MS 橙皮苷 橙皮素 柚皮素 原儿茶酸 去氢木香内酯 川陈皮素 去甲基川陈皮素 小檗碱 桔皮素 

分 类 号：R969.1[医药卫生—药理学]