目的 研究左乙拉西坦对甲氨蝶呤在大鼠体内吸收的影响及相关机制,为临床合理使用两药提供参考。方法 ①药动学研究:将体重(250±20) g SD大鼠随机分为甲氨蝶呤单用组和甲氨蝶呤与左乙拉西坦合用组,每组5只,灌胃给药,在设计时间点眼底静脉丛取血后离心取血浆用于检测。②体外翻转肠实验:取大鼠10 cm空肠并用毛细管将肠段轻柔翻转,结扎肠肛侧端,向翻转后的肠腔内加入1 mL KRB缓冲液后将肠囊垂直放入含药溶液。③Caco-2细胞摄取实验:Caco-2细胞接种于24孔板培养15 d后进行摄取实验,按实验分组给药孵育30 min后终止摄取并清洗,裂解细胞用于甲氨蝶呤与蛋白的检测并计算药物摄取量。结果 ①甲氨蝶呤在单用组与合用组的主要药动学参数如下:ρmax(ng·mL-1):28.6±3.04、54.33±13.97;tmax(min):50±8.32、70±7.32;AUC0-t(ng·min·mL-1):8 230±2 274、15 003±3 359; AUC0-∞(ng·min·mL-1):8 450±2 125、15 108±3 371;CL(mL-1·min·kg-1):161.80±27.60、90.43±13.76。合用左乙拉西坦后,甲氨蝶呤的ρmax、tmax、AUC0-t、AUC0-∞值升高,CL值下降,差异均有统计学意义。②与单用组相比,合用组的左乙拉西坦显著增加翻转肠浆膜侧甲氨蝶呤的药物浓度,其作用表现为时间依赖性。③维拉帕米、地高辛和左乙拉西坦均可增加甲氨蝶呤在Caco-2细胞的摄取,且左乙拉西坦的作用表现为剂量依赖性。结论 当甲氨蝶呤和左乙拉西坦联合应用时,可能会在肠道发生由外排型转运体P-糖蛋白所介导的药物相互作用,使甲氨蝶呤外排减少、吸收增加、血药浓度增加。提示临床上联合使用甲氨蝶呤和左乙拉西坦时需调整甲氨蝶呤的剂量,必要时进行血药浓度监测。
Abstract
OBJECTIVE To study the effect of levetiracetam on the absorption of methotrexate in rats and related mechanisms, and provide reference for clinical rational use of two drugs. METHODS ①Pharmacokinetic study: Sprague Dawley (SD) Rats (250±20)g were randomly divided into control group and experimental group, 5 rats in each group, administered by intragastric administration. At the design time point, blood was taken from the fundus venous plexus and centrifuged. Supernatant plasma for detection. ②The intestine experiment was performed in vitro: the rat 10 cm jejunum was taken and the intestinal segment was gently inverted with a capillary tube, and the intestinal anal side was ligated. After the 1 mL KRB buffer was added to the inverted intestinal lumen, the intestinal sac was placed vertically into the drug-containing solution. ③Caco-2 Cells uptake experiments: The Caco-2 cells were inoculated in a 24-well plate for 15 days and then subjected to an uptake experiment. After the incubation for 30 minutes, the cells were stopped for uptake and washed, and the cells were lysed for the detection of methotrexate and protein. Calculate the amount of drug intake. RESULTS ①The main pharmacokinetic parameters of control group and experimental group were as follows: ρmax(ng·mL-1): 28.6±3.04、54.33±13.97; tmax(min): 50±8.32、70±7.32; AUC0-t(ng·min·mL-1): 8 230±2 274、 15 003±3 359; AUC0-∞(ng·min·mL-1): 8 450±2 125、15 108±3 371; CL(mL-1·min·kg-1): 161.8±27.6、 90.43±13.76. In the experimental group, the ρmax, tmax, AUC0-t and AUC0-∞ values of methotrexate increased, and the CL value descreased. The difference was statistically significant. ②Compared with the control group, levetiracetam in the experimental group significantly increased the drug concentration of the methotrexate on the serosal side of the intestine, and its effect was time-dependent. ③Verapamil, digoxin and levetiracetam increased the uptake of methotrexate in the Caco-2 cells, and the effect of levetiracetam was dose-dependent. CONCLUSION When methotrexate and levetiracetam were combined, the drug interactions mediated by the efflux transporter P glycoprotein in the intestinal tract. Methotrexate reduces efflux in the body, increases absorption, and increases plasma concentration. It is suggested that the dose of methotrexate should be adjusted when the combination of methotrexate and levetiracetam, and blood concentration should be monitored if necessary.
关键词
左乙拉西坦 /
甲氨蝶呤 /
大鼠 /
P-糖蛋白 /
药动学
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Key words
levetiraceta /
methotrexate /
rat /
P-glycoprotein /
pharmacokinetics
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中图分类号：
R969.1
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参考文献
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脚注
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基金
河北省医学科学研究重点课题计划项目资助(20160062);河北省政府资助临床医学优秀人才培养项目资助(361003)
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