摘要
目的:研究炒莱菔子醇提物中芥子酸在正常大鼠体内的药代动力学特征以及在哮喘大鼠病理状态下的药代动力学-药效动力学(PK-PD)相关性。方法:正常大鼠分别单次灌胃低、中、高剂量(4.5,9,18 g·kg^-1)的炒莱菔子醇提物,利用UHPLC-MS/MS测定不同时间点血浆中芥子酸含量,采用DAS 3.2.8软件进行数据分析以获取药代动力学参数;采用腹腔注射卵清蛋白佐以氢氧化铝建立大鼠哮喘模型,分别灌胃低、高剂量(4.5,9 g·kg^-1)炒莱菔子醇提物,3周后眼底静脉丛取血,分离血浆和血清,UHPLC-MS/MS测定不同时间点血浆中芥子酸含量,利用酶联免疫吸附测定法(ELISA)测定血清中免疫球蛋白E(Ig E),白细胞介素-5(IL-5),肿瘤坏死因子-α(TNF-α)活性,采用DAS 3.2.8软件进行PK-PD模型拟合以及数据分析。结果:正常大鼠灌胃给予低、中、高剂量炒莱菔子醇提物后,血浆中芥子酸药峰浓度(Cmax)分别为(29.35±10.32),(62.70±27.47),(137.33±40.95)μg·L^-1;药时曲线下面积(AUC0-t)分别为(92.83±27.16),(240.74±75.09),(633.95±195.88)μg·L^-1·h;达峰时间(Tmax)分别为(2.58±0.80),(3.00±0),(5.50±1.23)h;与炒莱菔子醇提物低剂量组比,炒莱菔子醇提物中、高剂量组的AUC0-t,平均滞留时间(MRT0-t)均显著增加(P<0.05,P<0.01);芥子酸体内AUC0-t在4.5~18 g·kg-1线性依赖性良好。哮喘大鼠分别灌胃低、高剂量炒莱菔子醇提物后0.083,0.167 h,与模型组比较,血清中IL-5,Ig E,TNF-α含量均有不同程度地降低(P<0.05,P<0.01);血浆中芥子酸Cmax分别为(58.43±27.94),(61.16±18.79)μg·L^-1;AUC0-t分别为(188.75±37.07),(247.90±36.89)μg·L^-1·h;AUC0-t,表观分布容积(Vz/F)和清除率(CLz/F)均随剂量增加而增加,最佳药动学模型拟合为血管外给药的一房室模型,PK-PD模型可能适用于间接连接模型。结论:哮喘大鼠体内芥子酸血药浓度与血清中IL-5,Ig E,TNF-α含量存在一定的相关性,给药剂量和机能状态(病理或生理状态)均会影响炒莱菔子中芥子酸在大鼠体内的药代动力学行为,并与其平喘药效有一定的关联性。
Objective:To study the pharmacokinetics of sinapic acid from stir-fried Raphani Semen in normal rats and the correlation between pharmacokinetics-pharmacodynamics(PK-PD)in asthma rats.Method:Normal rats received 4.5,9,18 g·kg^-1 of stir-fried Raphani Semen by oral administration,respectively.Blood was taken from ophthalmic venous plexus at different time points according to the experimental design,the plasma concentration of sinapic acid was analyzed by UHPLC-MS/MS,and data analysis was performed using DAS 3.2.8 software to obtain the pharmacokinetic parameters.Rat asthma model was established by intraperitoneal injection of ovalbumin with aluminum hydroxide,and treated with ethanol extract of stir-fried Raphani Semen(low and high doses of 4.5,9 g·kg^-1).After treatment for 3 weeks,taking blood at different time points,plasma and serum were separated.UHPLC-MS/MS was established for the determination of plasma concentration of sinapic acid,contents of interleukin-5(IL-5),immunoglobuin E(IgE),tumor necrosis factor-α(TNF-α)in serum at different time points were detected by enzyme-linked immunosorbent assay(ELISA),DAS 3.2.8 software was used for PK-PD model fitting and data analysis.Result:After normal rats were administrated with low,medium and high doses of stir-fried Raphani Semen,the peak concentration(Cmax)of sinapic acid in plasma were(29.35±10.32),(62.70±27.47),(137.33±40.95)μg·L^-1,its area under the curve(AUC0-t)were(92.83±27.16),(240.74±75.09),(633.95±195.88)μg·L^-1·h,its peak time(Tmax)were(2.58±0.80),(3.00±0),(5.50±1.23)h,respectively.Compared with the low dose group,AUC0-t and mean retention time(MRT0-t)were all increased in the medium and high dose groups,showing statistical differences(P<0.05,P<0.01).The linear relationship of AUC0-t in sinapic acid was good within the dose range of 4.5-18 g·kg-1.After treating with ethanol extract of stir-fried Raphani Semen for 0.083,0.167 h,compared with the model group of asthmatic rats,serum levels of IL-5,IgE,TNF-αof the medication groups were decreased to different degrees(P<0.05,P<0.01).Cmax of sinapic acid in the low and high dose groups were(58.43±29.94),(61.16±18.79)μg·L^-1,its AUC0-twere(188.75±37.07),(247.90±36.89)μg·L^-1·h,respectively.AUC0-t,apparent volume of distribution(Vz/F)and clearance rate(CLz/F)all increased significantly with the increase of dose.The best pharmacokinetic model of sinapic acid was fitted as a one-compartment model for extravascular administration,PKPD model may be applicable to indirect connection model.Conclusion:The plasma concentration of sinapic acid is correlated with contents of IL-5,IgE and TNF-α,dosage and functional state(pathological or physiological state)can affect the pharmacokinetic behavior of sinapic acid from stir-fried Raphani Semen in rats,and it has a certain correlation with the anti-asthmatic effect.
作者
李丽
殷小杰
李丽
王岚
贡磊磊
梁日欣
LI Li;YIN Xiao-jie;LI Li;WANG Lan;GONG Lei-lei;LIANG Ri-xin(Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;Beijing Obstetrics and Gynecology Hospital,Capital Medical University,Beijing 100026,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2020年第7期109-116,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金面上项目(81473355).
