小鼠灌胃赤芍单味药和桃红四物汤后体内芍药苷的水平比较(英文)

Intragastric administration of Radix Paeoniae Rubra Extracts versus Taohong Siwu Tang for the serum concentration of peoniflorin in mice

背景:桃红四物汤由桃仁、红花、当归、赤芍、生地、川芎组成。芍药苷为赤芍单味药和中药复方桃红四物汤的主要有效成分之一,具有活血化瘀,抗急性心肌缺血的作用。目的:比较小鼠灌胃赤芍和桃红四物汤后血清中芍药苷的质量浓度。设计:随机对比观察。单位:河北北方学院药学系,河北北方学院基础医学部。材料:实验于2005-02/06在河北北方学院药学系第一实验室完成。选用18～22g昆明种小鼠80只,雌雄兼有,清洁级,购自北京中国科学院遗传与发育生物学研究所实验动物中心。6种中药桃仁、红花、当归、赤芍、生地、川芎均购自张家口中医研究院。方法:①取8只小鼠空腹12h后分离空白血清,于空白血清中加芍药苷对照品溶液,乙腈沉淀蛋白,离心分离,取上清液20μL,得回归方程。②将72只昆明种小鼠按随机数字表法分为赤芍组和桃红四物汤组,每组36只。空腹12h后,分别灌胃赤芍提取物及桃红四物汤提取物,以芍药苷计300mg/kg,灌胃后于15,30,60,90,120,180min取血1mL(每组每个时间点各6只),制备血清样品。比较小鼠灌胃赤芍提取物与桃红四物汤提取物后血清中芍药苷达峰时间和达峰浓度的差异。高效液相色谱法测定芍药苷含量,色谱条件为:色谱柱YWG-C18(250×4.6mm,10μm),流动相甲醇-0.05mol/LKH2PO4(38∶62);流速1.0mL/min;检测波长230nm;柱温25℃。主要观察指标:赤芍和桃红四物汤在小鼠血清中芍药苷的质量浓度。结果:72只小鼠用于赤芍和桃红四物汤实验,全部进入结果分析,无脱失。①色谱分离结果:芍药苷与血清中其他成分能很好分离,在5.40～646.0μg/mL范围内线性关系良好,平均回收率为93.6%,最低监测浓度1.08mg/L。②灌胃不同时间两组小鼠血清中芍药苷的质量浓度比较:赤芍组和桃红四物汤组分别在灌胃60,30min后达峰值(46.82±5.29),(36.27±5.72)mg/L,桃红四物汤组的峰值浓度低于赤芍组(P<0.01)。结论:复方配伍后,血清中芍药苷浓度明显降低,提示配伍组方后其他化学成分可能促进了芍药苷的代谢转化,从而降低了血清中原形药的浓度。

BACKGROUND： Taohong Siwu Tang is made up of Taoren, Honghua, Danggui, Chishao and Chuangxiong. Paeoniflorin is one of the major effective components in Radix Paeoniae Rubra Extracts and Taohang Siwu Tang, and it can promote blood circulation and remove blood stasis and has the effect of anti-acute myocardial ischemia. OBJECTIVE： To compare the serum concentration of paeoniflorin in mice after intragastric administration （i.g.） of Radix Paeoniae Rubra extracts and Taohong Siwu Tang. DESIGE： Rndomized controlled observation. SETTING： Department of Pharmacy and Department of Basic Medicine, Hebei North University. MATERIALS： The trial was performed in the first laboratory of Department of Pharmacy, Hebei North University during February to June 2005. Eighty involved Kunming mice of either gender and of clean grade, with body mass of 18 to 22 g, were purchased from Experimental Animal Center, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences. Taoren, Honghua, Danggui, Chishao, Shengdi and Chuanxiong 6 Chinese herbs were all purchased from Zhangjiakou Institute of Traditional Chinese Medicine. METHODS：①Blank serum was isolated from eight mice which were fasted for 12 hours. Control sample solution of paeoniflorin was added into blank serum. Acetonitrile was used to deposit protein, the solution was centrifugated and isolated and 20μL supernatant was taken to obtain regression equation. ② Seventy-two Kunming mice were randomized into Radix Paeoniae Rubra extracts group and Taohong Siwu Tang group, with 36 mice in each. After being fasted for 12 hours, the mice in each group were intragastric administration of Radix Paeoniae Rubra extracts and Taohong Siwu Tang, respectively and the concentration of paeoniflorin was 300 mg/kg. 1 mL blood was taken respectively at 15, 30, 60, 90, 120 and 180 minutes after intragastric administration and prepared into serum sample （6 mice at each time point in each group）. The time to peak and difference in peak concentration of serum paeoniflorin were compared between intragastric administration of Radix Paeoniae Rubra extracts and Toohong Siwu Tang. The concentration of paeoniflorin was measured with high-performance liquid chromatography （HPLC）. Chromatographic condition： chromatographic column YWG-C18 （250×4.6 mm,10μm）. The mobile phase was methanolKH2PO4 （0.05 mol/L） （38：62） at a flow rate of 1.0 mL/min. The detective wavelength was set at 230 nm. The column temperature was 25 . MAIN OUTCOME MEASURES： Serum concentration of paeoniflorin after administration of Radix Paeoniae Rubra extracts and Taohong Siwu Tang in mice. RESULTS： Seventy-two mice were used for Radix Paeoniae Rubra extr acts and Taohong Siwu Tang tests, and all of them entered the stage of result analysis without deletion, ① Chromatographic isolation results： Paeoniflorin could be well isolated from other components. The linear range for detection of paeoniflorin was 5.40-646.0μg/mL. The mean recovery rate was 93.6% and the limit of detection was determined as 1.08 mg/L. Comparison of concentration of serUm paeoniflorin of mice between two groups： The concentration of serum paeoniflorin in the Radix Paeoniae Rubra extracts group and Taohang Siwu Tang group reached the peak at 30 and 60 minutes after intragastric administration, respectively. The peak concentration of paeoniflorin of Taohong Siwa Tang group was lower than that of Radix Paeoniae Rubra extracts group [（36.27±5.72） mg/L vs. （46.82±5.29） mg/L, P 〈 0.01 ）]. CONCLUSION： The concentration of serum paeoniflorin is significantly decreased after intragastric administration, It was further confirmed the decreased concentration of the crude paeoniflorin in the mice sera might result from promotion of its metabolic conversion by other chemical components in the complex prescription. This result needs further investigation,