葛根素在大鼠体内的药代动力学特征和安全性的研究

Study on pharmacokinetic characteristics and safety of puerarin in rats

目的评价葛根素在大鼠体内的药代动力学特征和安全性,为葛根素的临床用药方案提供参考。方法色谱柱Agilent proshell 120 EC-C18柱(3 mm×100 mm,2.7μm);流动相乙腈(B)-水(A),梯度洗脱,流速0.3 mL·min^(-1);柱温30℃。每组5只SD大鼠分别灌胃给药低剂量葛根素(100 mg·kg^(-1))和高剂量葛根素(400 mg·kg^(-1)),并于给药0、5、10、15、30、60、90、120、180、240、360、480和600 min后经眼眶静脉丛采血约0.5 mL,使用高效液相色谱法(HPLC)检测血药浓度,用DAS 2.0软件算出葛根素在SD大鼠体内的药动学参数。另外,高剂量葛根素(400 mg·kg^(-1))灌胃4周后,检测SD大鼠肝功能指标谷草转氨酶(GOT)和谷丙转氨酶(GPT)、肾功能指标血肌酸酐(SCr)和尿素氮(BUN)的变化。结果葛根素的线性范围为1.875~7680.00μg·mL^(-1),回收率、精密度和稳定性均良好且达到测定要求。低剂量葛根素组t_(max)、C_(max)、t_(1/2)、t_(1/2)β、AUC_(0-t)、AUC_(0-∞)、MRT_(0-t)、MRT_(0-∞)分别为(0.38±0.16)h、(1.40±0.37)μg·mL^(-1)、(0.53±0.47)h、(0.78±0.28)h、(1.97±1.11)mg·h·L^(-1)、(2.02±1.13)mg·h·L^(-1)、(1.17±0.06)min、(1.30±0.13)min,高剂量葛根素组tmax、Cmax、t1/2、t1/2β、AUC_(0-t)、AUC_(0-∞)、MRT_(0-t)、MRT_(0-∞)分别为(0.67±0.31)h、(2.79±1.26)μg·mL^(-1)、(0.48±0.10)h、(0.95±0.25)h、(4.30±0.58)mg·h·L^(-1)、(4.31±0.57)mg·h·L^(-1)、(1.26±0.36)min、(1.30±0.36)min;高剂量葛根素组的C_(max)、AUC_(0-t)明显高于低剂量葛根素组。高剂量葛根素组的GOT、GPT、SCr和BUN水平未见明显变化。结论口服高剂量葛根素在大鼠体内吸收好,且安全性良好。

Objective To evaluate the pharmacokinetic characteristics and safety of puerarin in rats and to provide a reference for formulation and selection of clinical puerarin medication scheme.Methods Chromatographic column:Agilent proshell 120 EC-C18 column(3 mm×100 mm,2.7μm);mobile phase acetonitrile(B)-water(A),gradient elution,flow rate 0.3 mL min^(-1);column temperature 30℃.SD rats(n=5 in each group)were given low-dose puerarin(100 mg·kg^(-1))and high-dose puerarin(400 mg·kg^(-1))by intragastric administration.Blood(0.5 mL)was collected through the orbital venous plexus after treatment with puerarin at 0,5,10,15,30,60,90,120,180,240,360,480 and 600 min.The blood drug concentration was detected by high performance liquid chromatography(HPLC),and the pharmacokinetic parameters were calculated by DAS 2.0 software.In addition,after gavage of high-dose puerarin(400 mg·kg^(-1))for 4 weeks,the liver function indexes of glutamic oxalacetic transaminase(GOT)and glutamic-pyruvic transaminase(GPT),the renal function indexes of serum creatinine(SCr)and urea nitrogen(BUN)in SD rats were detected.Results The linear range of puerarin was 1.875-7680.00μg·mL^(-1),the recovery rate,precision and stability were good and met the determination requirements.The pharmacokinetic parameters t_(max),C_(max),t_(1/2),t_(1/2)β,AUC_(0-t),AUC_(0-∞),MRT_(0-t),MRT_(0-∞)were(0.38±0.16)h,(1.40±0.37)μg·mL^(-1),(0.53±0.47)h,(0.78±0.28)h,(1.97±1.11)mg·h·L^(-1),(2.02±1.13)mg·h·L^(-1),(1.17±0.06)min,(1.30±0.13)min in low-dose puerarin group,while which were(0.67±0.31)h,(2.79±1.26)μg·mL^(-1),(0.48±0.10)h,(0.95±0.25)h,(4.30±0.58)mg·h·L^(-1),(4.31±0.57)mg·h·L^(-1),(1.26±0.36)min,(1.30±0.36)min in high-dose puerarin group.C_(max)and AUC_(0-t)of the high-dose puerarin group were significantly higher than those of the low-dose puerarin group.After 4 weeks of high-dose puerarin intervention,the plasma levels of GOT,GPT,SCr and BUN did not change significantly,indicating that the oral dose is safe.Conclusion Oral high-dose puerarin is well absorbed and safe in rats.