基于肝微粒体代谢的盐酸小檗胺对盐酸小檗碱的酶动力学研究  

Study on the enzyme kinetics of berbamine hydrochloride on berberine hydrochloride based on metabolism of liver microsomes

作　　者：毕若红 冯慧 潘琳 高洁 黎世玲 唐茜 赖先荣[1] BI Ruo-hong;FENG Hui;PAN Lin;GAO Jie;LI Shi-ling;TANG Xi;LAI Xian-rong(School of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,Sichuan)

机构地区：[1]成都中医药大学药学院,四川成都611137

出　　处：《中药与临床》2021年第4期31-35,共5页Pharmacy and Clinics of Chinese Materia Medica

摘　　要：目的:基于高效液相色谱法建立底物盐酸小檗碱的定量分析方法,研究盐酸小檗胺对盐酸小檗碱在大鼠和人肝微粒体中代谢的酶动力学特征。方法:考察酶反应体系中孵育时间、肝微粒体蛋白浓度、盐酸小檗碱浓度以及盐酸小檗胺对盐酸小檗碱代谢反应的影响,并采用双倒数作图法计算盐酸小檗碱的酶促反应动力学参数Km和Vmax。结果:盐酸小檗碱在1-25 μg-mL^(-1)范围内有良好的线性关系。日内、日间精密度均符合要求,平均回收率在96.61%-97.55%,室温、4℃冰箱中放置24 h下稳定性良好;在此基础上确定肝微粒体蛋白浓度为0.5 g.L^(-1),盐酸小檗碱质量浓度为2.5-20 μg·mL^(-1)时,在37℃,120 r·min^(-1)条件下孵育60 min为最佳孵育条件。同时计算酶动力学参数,大鼠肝微粒体中的Km为1.6157 μg·mL^(-1),Vmax的值 12.8535μg.g-1.min^(-1),人肝微粒体的Km值为 1.4775 μg.mL^(-1),Vmax值 14.9701 μg.g-1·mL^(-1)。结论:盐酸小檗胺与盐酸小檗碱在适宜的比例及浓度范围内,盐酸小檗胺能够促进盐酸小檗碱在大鼠和人肝微粒体中的代谢速率。Objective:The quantitative analysis method of berberine hydrochloride based on high performance liquid chromatography(HPLC) was established to study enzyme kinetic characteristics of berbamine hydrochloride metabolism in rat and human liver microsomes.Method:The effects of incubation time,liver microsome protein concentration,berberine hydrochloride concentration and the effect of berbamine hydrochloride on the metabolic rate of berberine hydrochloride were investigated.The kinetic parameters Km and Vmax of the enzymatic reaction of berberine hydrochloride were calculated by Lineweaver-Burk plot.Result:The linear range of berberine hydrochloride was 1~25 μg·mL^(-1).The precision of intra-day and intra-day met the requirement.The average recoveries were in the range of 96.61% ~97.55%.Berberine hydrochloride was stable after 24 h storage at room temperature and 4℃.On this basis,the optimal incubation conditions were determined as follows:berberine hydrochloride was incubated at 37℃ and 120 r·min^(-1) for 60 min when the concentration of liver microsomal protein was 0.5 g·L-1,and the concentration of berberine hydrochloride was 2.5~20 μg·mL^(-1).At the same time,the kinetic parameters of enzyme were calculated.In rat liver microsomes,Km was 1.6157 μg·mL^(-1) and Vmax was 12.8535 μg·g-1·min^(-1).In human liver microsomes,Km value was 1.4775 μg·mL^(-1) and value was 14.9701 μg·g-1·min^(-1).Conclusion:Berbamine hydrochloride can promote the metabolic rate of berberine hydrochloride in rat and human liver microsomes in appropriate ratio and concentration range.

关 键 词：糖尿病 盐酸小檗碱 盐酸小檗胺 肝微粒体代谢 酶促动力学 

分 类 号：R285.5[医药卫生—中药学]