玳玳果黄酮降脂提取物效应组分大鼠肝肠微粒体代谢特性研究

Metabolic characteristics of active parts of lipid-lowering flavonoid extract of Daidai in liver and intestinal microsomes of rats

该文比较玳玳果黄酮降脂提取物效应组分新橙皮苷、柚皮苷在大鼠肝脏及肠道微粒体中的代谢差异,探讨效应组分在肝脏及肠道的Ⅰ相代谢特性。采用UPLC-MS/MS技术建立效应组分在肝肠微粒体Ⅰ相代谢孵育体系中的定量分析方法,并以差速离心法制备大鼠肝、肠微粒体,构建Ⅰ相代谢孵育体系,通过将各时间点药物剩余百分率的自然对数ln(X)对时间t作图计算代谢消除半衰期,比较效应组分的代谢特征。结果表明,所建立的UPLC-MS/MS定量分析方法专属性、标准曲线及线性范围良好,方法准确度与精密度、定量下限均符合规定,该方法能够满足大鼠肝肠微粒体Ⅰ相代谢孵育体系下效应组分的定量检测需要;大鼠肝微粒体Ⅰ相孵育体系中新橙皮苷代谢消除半衰期为(2. 20±0. 28) h,大于柚皮苷的底物消除半衰期(1. 97±0. 28) h,但差异无统计学意义;大鼠肠微粒体Ⅰ相孵育体系中新橙皮苷的代谢消除半衰期为(3. 68±0. 54) h,大于柚皮苷的底物消除半衰期(2. 26±0. 13) h,且差异有统计学意义(P<0. 05);提取物效应组分在肝微粒体中的消除半衰期均小于肠微粒体。该研究发现,玳玳果黄酮降脂提取物效应组分在大鼠肝、肠微粒体Ⅰ相代谢孵育体系中具有不同的消除半衰期,提示二者在大鼠肝脏及肠道中具有不同的代谢特征,肝脏可能为玳玳果黄酮降脂提取物效应组分的主要代谢部位,且提取物效应组分柚皮苷的Ⅰ相代谢强度大于新橙皮苷的代谢强度,推测导致该代谢特征差异的原因可能与黄酮苷B环羟基结合位点及甲氧基数量有关。为进一步指导玳玳果黄酮降脂提取物制剂的深度开发及临床开发应用等提供重要的实验依据。

The paper studies and compares the metabolic difference of active ingredients of lipid-lowering flavonoid extract of Daidai in rat livers and intestinal microsomes, in order to explore the phase Ⅰ metabolism characteristics of active ingredients in livers and intestines. UPLC-MS/MS was used to establish a quantitative analysis method for active ingredients, neohesperidin and narngin, in a phase Ⅰ metabolism incubation system of liver and intestinal microsomes. Differential centrifugation was used to make liver and intestinal microsomes of rats. A phase Ⅰ metabolism incubation system was established, and the concentrations of the residual at different incubation time points were analyzed. Graphs were plotted to calculate the metabolic elimination half-life of the main active parts, with the natural logarithm residual percentage values In(X) at different time points as the y axis, and time t as the x axis. The metabolism characteristics of the active ingredients were compared. The established UPLC-MS/MS quantitative analysis method has a good specialization ,standard curve and linear range, accuracy and precision, with a satisfactory lower quantitative limit. The method allows quantitative detection of the active ingredients in a phase Ⅰ metabolism incubation system of liver and intestinal microsomes of rats. In the rats liver microsomes incubation system, the metabolic elimination half-life of neohesperidin and narngin were ( 2. 20 ±0. 28) h and (1. 97±0. 28) h respectively. The elimination half-life of neohesperidin was larger than that of narngin, but with no statistically significant difference. In the rats intestinal microsomes incubation system, the metabolic elimination half-lives of neohesperidin and narngin were ( 3. 68±0. 54) h and (2. 26±0. 13) h respectively. The elimination half-life of neohesperidin was larger than that of narngin, with statistically significant differences ( P<0. 05). The elimination half-lives of the active ingredients in liver microsomes were smaller than those in intestinal microsomes. The experiment results showed that the active ingredients of lipid-lowering flavonoid extract of Daidai had different elimination half-lives in phase Ⅰ rats liver and intestinal microsomes incubation system. This implied that they had different metabolic characteristics in rats liver and intestine, and liver may be the main metabolism site of the active ingredients. The phase Ⅰ metabolism of narngin was stronger than that of neohesperidin. the differences between their metabolic characteristics may be related to the binding sites of B-ring hydroxyl in flavonoid glycosides and the number of methoxyl group. The results provided an important experimental basis for further development and clinical application of lipid-lowering flavonoid extract preparation of Daidai.