清开灵注射液中非胆酸成分对胆酸和异去氧胆酸药代动力学影响

Effects of the Part of Non-Cholic Acid in Qingkailing Injection on Pharmacokinetics of Cholic Acid and Hyodeoxycholic Acid

目的研究清开灵注射液中非胆酸成分对胆酸(CA)和异去氧胆酸(HDCA)药代动力学过程及在主要脏器分布的影响,以阐明清开灵注射液的药理作用。方法实验动物分2组,清开灵注射液(10ml.kg-1)组和单用相同剂量的CA/HDCA组,给药前小鼠禁食不禁水12h,眼眶微量采血20μl,直接沉淀蛋白振荡后离心,取上清进样检测。取心、肺、肾、脑清洗后用滤纸吸干称量,用1∶4匀浆后离心取上清,处理方法同血样进样检测。使用液相色谱/质谱联用法检测血和组织样品中的CA和HDCA的含量。结果CA和HDCA在全血和组织中的药物浓度线性范围为4～4000μg.L-1,线性关系良好(r>0.999),最低检测浓度为2μg.L-1,日间和日内精密度的RSD均<5%。比较复方中和单用时CA和HDCA的药代动力学参数和在主要组织(心、肺、肾、脑)中的分布,清开灵注射液中的CA和HDCA的AUC、MRT、T1/2β、V均明显高于单用时(P<0.05),CL明显低于单用时(P<0.05);清开灵注射液中CA和HDCA的分布明显优于单用CA/HDCA,CA和HDCA在组织中的分布特点为:C肺>C肾>C心>C脑。结论本方法灵敏度高,特异性好,适用于胆酸类化合物的体内过程研究;清开灵注射液中非胆酸成分可延长胆酸成分在体内的滞留时间和促进胆酸成分在组织中的分布;CA和HDCA在脑中浓度低说明其不易透过血脑屏障;复方制剂中多种成分存在相互作用。

Aim To study pharmacokinetics and tissue distribution of cholic acid（CA） and hyodeoxycholic acid（ HDCA） in Qingkailing Injection and to explore the effects of the part of non-cholic acid in Qingkailing Injection on pharmacokinetics of CA and HDCA. Mehtods Mice were randomly divided into the Qingkailing Injection group（ 10 ml·kg^-1）and CA/HDCA group （at the same dosage as in Qingkailing Injection）. The concentrations of CA and HDCA in mice blood and tissues were measured by LC/ESI/MS. Results The standard curves of CA and HDCA in blood and tissues were linear in the range of 4 -4 000μg·L^-1 （ r 〉0.999）, the lowest limit of quantitation being 2μg·L^-1. There were significant differences （ P 〈0.05） between the Qingkailing Injection group and the CA/HDCA group in main parameters （AUC,MRT, T1/2β ,V and CL ） and distributions in heart, lung, kidney and brain. Conclusion The method is simple, rapid, sensitive,reliable and is applicable to assay CA and HDCA in biological samples. The parts of non-cholic acid in Qingkailing Injection can extend MRT and improve distribution of CA and HDCA. CA and HDCA can＇ t permeate blood brain barriers easily. The result showed that there were interactions between ingredients in compound preparations.