目的：黄连碱是黄连中一种主要的异喹啉类生物碱，具有较好的抗糖尿病药理活性。为了探讨黄连碱在正常和糖尿病大鼠体内的代谢产物及其代谢途径是否存在差异。方法：本实验采用SD大鼠禁食12h后腹腔注射链脲佐菌素（STZ）制备糖尿病大鼠模型。造模成功后分别单次灌胃正常和糖尿病大鼠黄连碱（剂量20 mg/kg），收集给药后0~48 h的尿液和粪便以及0~36 h的胆汁样品，采用高分辨HPLC-MS/MS技术对生物样品中的药物原型及代谢物进行鉴定。选用 Agilent TC-C18 色谱柱（4.6 mm×150 mm，5μm），以乙腈- 0.1%甲酸水溶液为流动相梯度洗脱，流速 1.0 mL·min-1，柱温 25 ℃。质谱使用电喷雾离子源（ESI），正、负离子检测模式，扫描范围 m/z 50~700。对各色谱峰质谱图进行分析，根据准分子离子峰判断相对分子质量，进一步根据各色谱峰的主要碎片离子、保留时间等信息以及相关文献数据，推测化合物的结构。结果：黄连碱在正常和糖尿病大鼠体内可发生较广泛的Ⅰ相和Ⅱ相代谢，在大鼠尿液、粪便、胆汁共鉴定出14个代谢产物，其中尿液10中个、粪便中11个、胆汁中8个；Ⅰ相代谢物5个，主要为羟基化物和去甲基化物，Ⅱ相代谢物9个，主要为葡萄糖醛酸化物，但由COP脱氢后生成的M1再次硫酸酯化的产物M4在糖尿病大鼠胆汁中未检出。结论：我们推测M1在糖尿病和正常大鼠胆汁中代谢过程的差异是黄连碱产生降糖作用的关键，但其他代谢产物量上的差异有待进一步研究确定。本研究可为黄连碱的药效学和药理学研究提供一定的物质基础。
Objective:Coptisine is one of the main isoquinoline alkaloids in rhizoma coptidis，which has good anti-diabetic pharmacological activity.To investigate whether there are differences in metabolites and metabolic pathways of coptidine in normal and diabetic rats. Methods: In this study, the diabetic rat model was prepared by intraperitoneal injection of STZ after 12h fasting.After successful modeling, the rats of normal and diabetic rats were dosed with berberine (20 mg / kg), and urine and feces from 0 to 48 h and bile samples from 0 to 36 h were collected.The drug prototypes and metabolites in the biological samples were identified by high resolution HPLC-MS /MS technique.Agilent TC-C18 column (4.6 mm×150 mm,5μm) was selected, useing acetonitrile-0.1% formic acid solution as mobile phase gradient elution, the flow rate was 1.0ml ·min-1, and the column temperature was 25℃. Mass spectrometry uses electrospray ion source (ESI), positive and negative ion detection mode, scanning range m/z 50~700.The mass spectrum of each chromatographic peak was analyzed, and the relative molecular mass was determined based on the excimer ion peak.The structure of the compound was inferred based on information such as the major fragment ions and retention times of each chromatographic peak and related literature data.Results:Coptisine can undergo extensive phase Ⅰ and phase Ⅱ metabolism in normal and diabetic rats.A total of 14 metabolites were identified in rat urine, stool, and bile. Among them, 10 were in urine, 11 were in feces, and 8 were in bile. There were 5 phase I metabolites, mainly hydroxyl and demethylates, and 9 phase II metabolites, mainly glucuronide. However, M4, the product of M1 resulfate after COP dehydrogenation, was not detected in bile of diabetic rats.Conclusion:We speculate that the difference in the metabolic processes of M1 in bile of diabetic and normal rats is the key to the hypoglycemic effect of berberine, but the differences in the amount of other metabolites need to be determined by further research. This study can provide a certain material basis for the pharmacodynamic and pharmacological studies of berberine.