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目的:探索补阳还五汤的主要成分在体内的代谢途径。方法:采用HPLC法对补阳还五汤中主要成分(毛蕊异黄酮苷、刺芒柄花苷、芍药苷、毛蕊异黄酮和刺芒柄花素)的含量进行测定,通过体外孵育方法考察补阳还五汤中主要成分在肝匀浆、肠道菌液、人工胃液和人工肠液中不同时间点的含量变化,以相对浓度的改变来反映各化合物量的变化。硝苯地平用来验证孵育体系中CYP450酶的活性。采用Shimadzu VP-ODS分析柱(250 mm×4.6 mm,5μm)色谱柱,柱温30℃,以0.1%甲酸水溶液(A)-乙腈(B)为流动相,梯度洗脱,流速0.8 m L·min～(-1),UV检测器波长254 nm,二极管阵列检测器检测波长范围200～500 nm,进样量10μL。结果:补阳还五汤中的5个主要成分在肝匀浆、肠道菌液、人工胃液和人工肠液中相对浓度均发生不同程度的改变,其中肝匀浆(毛蕊异黄酮含量增加超过500%)和肠道菌液(毛蕊异黄酮含量增加超过200%)孵育体系中相对浓度变化最为明显。结论:肠道菌群、消化道酶和肝药酶是补阳还五汤中主要成分的主要代谢因素,不同活性成分的主要代谢途径存在明显差异。
Objective: To explore the main components of Buyang Huanwu Decoction in vivo metabolic pathway. Methods: The contents of major constituents in BYHWD (Corydalis villosum, Thymus palmatum, Paeoniflorin, Calendula officinalis and Thorlastonin) in Buyang Huanwu Decoction were determined by HPLC. The main components of Wu Tang in liver homogenates, intestinal bacteria, artificial gastric juice and artificial intestinal fluid content changes at different time points, the relative concentration changes to reflect changes in the amount of each compound. Nifedipine was used to verify the activity of the CYP450 enzyme in the incubation system. A column with a Shimadzu VP-ODS analytical column (250 mm × 4.6 mm, 5 μm) was used. The column temperature was 30 ℃ and the mobile phase consisted of 0.1% formic acid in water (A) min ~ (-1), UV detector wavelength 254 nm, diode array detector detection wavelength range 200 ~ 500 nm, injection volume 10 μL. Results: The relative concentrations of five major components in Buyang Huanwu Decoction varied in varying degrees in the liver homogenates, gut bacteria, artificial gastric juice and artificial intestinal fluid. The liver homogenate (calycium isoflavone increased more than 500% ) And intestinal bacteria (calycosin content increased more than 200%) the relative concentration of the incubation system changes the most obvious. CONCLUSION: Intestinal flora, digestive tract enzyme and hepatic drug are the main metabolic factors of the main components in Buyang Huanwu Decoction. The main metabolic pathways of different active ingredients are obviously different.