基于远志酮在记忆障碍模型大鼠体内药代动力学特性的远志及开心散药物属性研究

Study on medical attribute of Yuanzhi(Radix Polygalae) and Kaixin San based on pharmacokinetics features of polygalaxanthone Ⅲ in rats with dysmnesia

目的建立RP-HPLC测定大鼠血清中远志酮Ⅲ浓度的方法,考量远志及其经典方开心散中远志酮Ⅲ在记忆障碍模型大鼠体内的药动学特性,研究远志及其复方在治疗痴呆中的药物属性。方法大鼠腹腔注射东莨菪碱致记忆障碍模型,分别灌胃给予远志水提物和开心散水提物,腹主动脉采血,离心,取血清适量,加0.1 mmol磷酸二氢钾-乙腈沉淀蛋白,取上清液氮气吹干,水溶解,过滤,用HPLC分析,以C18为固定相,流动相为乙腈-0.1%磷酸水溶液梯度洗脱,在254 nm检测远志酮Ⅲ血药浓度,Kinetica 4.4软件处理数据。结果远志酮Ⅲ血清在0.041 3～1.65 mg/L线性关系良好,最低定量限为41.3μg/L。记忆障碍模型大鼠灌胃给予远志水提物和开心散水提物后的远志酮Ⅲ药-时曲线均使用非房室模型处理,远志酮Ⅲ主要药动学参数曲线下面积(AUC),平均滞留时间(MRT)在远志提取物和开心散组间有差异(P<0.05)。口服开心散全方和远志提取物后远志酮Ⅲ呈现双峰吸收,达峰时间分别为15 min和150 min、15 min和180 min,口服开心散全方的远志酮ⅢAUC值是单味药远志的1.20倍,MRT是单味药远志的1.30倍。结论本试验建立的RP-HPLC测定法,专属、准确、灵敏,适用于远志酮Ⅲ在大鼠体内的药动学研究。大鼠口服远志药味提取物及其复方开心散提取物后远志酮Ⅲ的药代动力学结果表明通过复方配伍,可在提高生物利用度、加速吸收、延长有效血药浓度时间诸方面,调节其药动学特性,从而更有利于发挥其药效作用。

Objective To establish a method to determine the concentration of serum polygalaxanthone Ⅲ in rats,observe the pharmacokinetic features of Yuanzhi（Radix Polygalae） and polygalaxanthone Ⅲ in classic formula-Kaixin San in rats with dysmnesia,and study the medicinal properties of Yuanzhi and its compound formulas in the treatment of dysmnesia.Methods The model of dysmnesia was established by intraperitoneal injection of scopolamine in rats,and then the rats were intragastrically given the aqueous extracts of Yuanzhi and Kainxin San respectively.The blood samples were collected form rat abdominal aortic for separating serum samples with centrifugation.Monopotassium phosphate（0.1 mmol） and acetonitrile were added to deposit proteins,and then supernatant was collected,dried under nitrogen,dissolved in water,filtrated and analysed with HPLC.The concentration of polygalaxanthone Ⅲ was detected taking C18 as stationary phase and acetonitrile-0.1 % phosphoric acid solution as mobile phase in gradient elution with 254 nm detection wavelength.The data was treated with Kinetics 4.4 software.Results The medical serum of polygalaxanthone Ⅲ showed a good linear relationship at a range from 0.0413 mg/L to 1.65 mg/L.The lowest limit of quantification was 41.3 μg/L.The curve of medicinal-time of polygalaxanthone Ⅲ was treated by using non-compartment model after intragastrical giving of the aqueous extracts of Yuanzhi and Kainxin San.There was a difference in pharmacokinetic parameters AUC and MRT between extract of Yuanzhi and extract of polygalaxanthone Ⅲ（P0.05）.After orally giving whole formula of Kaixin San and extract of Yuanzhi,polygalaxanthone Ⅲ showed a bimodal absorption and the peaking time was,respectively,15 min and 150 min,and 15 min and 180 min.The value of AUC of polygalaxanthone Ⅲ was 1.20 times and MRT was 1.30 times higher than those of Yuanzhi after orally giving whole Kaixin San.Conclusion RP-HPLC determination established in this study is exclusive,accurate and sensitive,which is suitable for the pharmacokinetic research of polygalaxanthone Ⅲ in rats.The results of pharmacokinetic research of polygalaxanthone Ⅲ in rats after orally giving whole formula of Kaixin San and extract of Yuanzhi indicate that the combinations of compound formula can improve bioavailability,absorption and effective time of medicinal serum concentration,regulate the pharmacokinetic features and promote the effect.