基于组分配伍的马钱子碱与芍药苷经皮给药大鼠体内药代动力学及脑组织分布研究

Pharmacokinetics and brain distribution of brucine combined with paeoniflorin in rats after percutaneous administration based on component compatibility

目的:建立快速灵敏的LC-MS/MS分析方法同时测定大鼠血浆及脑组织中马钱子碱及芍药苷的含量,比较芍药苷与正常剂量及毒性剂量下马钱子碱配伍经皮给药前后大鼠体内药物动力学行为及脑组织分布情况。方法:以苦参碱为内标,流动相为乙腈-水(含10mmol/L乙酸铵+0.04%甲酸),梯度洗脱,对待测物进行分离。将大鼠随机分为6组,马钱子碱正常剂量组(18mg/kg)、芍药苷-1组(180mg/kg)、马钱子碱正常剂量-180mg/kg芍药苷配伍组、马钱子碱毒性剂量组(45mg/kg)、芍药苷-2组(450mg/kg)、马钱子碱毒性剂量-450mg/kg芍药苷配伍组,给药后分别于眼眶后静脉丛取血测定血药浓度及于2、6、12、24h处死大鼠,迅速分离大鼠脑组织,测定组织浓度。结果:与马钱子碱毒性剂量组比较,马钱子碱毒性剂量-450mg/kg芍药苷配伍组中马钱子碱的AUC_((0-t))、AUC_((0-∞))、C_(max)均显著减小(P<0.01,P<0.05),分别下降了45.79%、48.40%、49.70%,马钱子碱的t_(1/2z)及MRT均有所延长;配伍组中芍药苷的AUC_((0-t))、AUC_((0-∞))、C_(max)均显著增大(P<0.05,P<0.01),分别增加了64.66%、67.78%、85.96%。与马钱子碱正常剂量组比较,马钱子碱正常剂量组-180mg/kg芍药苷配伍组中马钱子碱的C_(max)、AUC_((0-t))及AUC_((0-∞))均有所减少,t_(1/2z)和MRT均有所延长,但无显著性差异;配伍组中芍药苷C_(max)显著增加(P<0.01),AUC_((0-t))及AUC_((0-∞))较配伍前分别增大了60.32%及49.61%,表明配伍极大地促进了芍药苷的经皮吸收。除24h外,配伍后马钱子碱脑部含量均有所下降,芍药苷脑组织浓度均有所上升。结论:芍药苷与正常剂量及毒性剂量下马钱子碱配伍,可抑制马钱子碱的经皮吸收,同时促进芍药苷的经皮渗透,具有一定的“增效减毒”效应,但具体的吸收机制还有待进一步深入研究。

Objective:To establish a rapid and sensitive LC-MS/MS analysis method for simultaneous determination the contents of brucine and paeoniflorin in plasma and brain tissues of rats,and then to compare the pharmacokinetic behavior and brain tissue distribution of rats before and after the combination of brucine with normal dose and toxic dose under percutaneous administration.Methods:The target compounds were eluted by gradient with matrine as internal standard and acetonitrile-water as mobile phase(containing 10mmol/L ammonium acetate+0.04%formic acid).Rats were randomly divided into 6 groups:normal dose of brucine group(18mg/kg),paeoniflorin-1 group(180mg/kg),normal dose of brucine and 180mg/kg paeoniflorin compatibility group,toxic dose of brucine group(45mg/kg),paeoniflorin-2 group(450mg/kg),and toxic dose of brucine and 450mg/kg paeoniflorin compatibility group.Blood samples were collected from the posterior orbital venous plexus in each group to determine the blood drug concentration,and the rats were sacrificed at 2h,6h,12h and 24h after administration,and the brain tissues of the rats were rapidly isolated to determine the tissue concentration.Results:The AUC_((0-t)),AUC_((0-∞))and C_(max) of brucine in the compatibility group decreased significantly(P<0.05)compared with the brucine toxicity dose group,which decreased by 45.79%,48.40%and 49.70%respectively.The t_(1/2) and MRT of brucine were prolonged,and the AUC_((0-t)),AUC_((0-∞))and C_(max) of paeoniflorin in the compatibility group were significantly increased(P<0.05),which increased by 64.66%,67.78%and 85.96%,respectively.The AUC_((0-t)),AUC_((0-∞))and C_(max) of brucine in the compatibility group decreased compared with the brucine normal dose group,and both t_(1/2) and MRT were extended,but there was no significant difference between the data.In the compatibility group,the C_(max) of paeoniflorin increased significantly(P<0.01),and the AUC_((0-t))and AUC_((0-∞))increased by 60.32%and 49.61%respectively compared with those before the compatibility,indicating that the compatibility greatly promoted the percutaneous absorption of paeoniflorin.In addition to the results of the 24 hours,the brain content of brucine decreased and the brain concentration of paeoniflorin increased after compatibility.Conclusion:The combination of strychnine and paeoniflorin under normal dose and toxic dose can inhibit the percutaneous absorption of strychnine and promote the percutaneous penetration of paeoniflorin.The compatibility of the two may have a certain‘synergistic and detoxifying’effect,but the related mechanism needs further study.