黄连素对CYP3A的体内外作用及相关性研究  

Effects of berberine hydrochloride on CYP3A and its correlation in vitro and in vivo

作　　者：唐霞[1,2] 张茂[1] 欧阳萌[2] TANG Xia;ZHANG Mao;OUYANG Meng(Department of Pharmacy,the First People s Hospital of Yibin City,Sichuan Province,Yibin 644000,China;Department of Clinical Pharmacology,Wuhan General Hospital of PLA,Wuhan 430070,China)

机构地区：[1]四川省宜宾市第一人民医院药学部,宜宾644000 [2]中国人民解放军武汉总医院临床药理科,武汉430070

出　　处：《西北药学杂志》2022年第1期45-50,共6页Northwest Pharmaceutical Journal

摘　　要：目的通过体外研究黄连素(BBR)对人肝癌细胞株HepG_(2)肝药酶CYP3A4在mRNA及蛋白水平表达的影响和在大鼠体内研究BBR对大鼠肝药酶CYP3A的底物咪达唑仑(MDZ)的药物代谢动力学(药动学)的影响,初步探讨BBR对CYP3A的体内外作用以及其是否存在相关性。方法体外实验用系列质量浓度BBR组、空白对照组分别与对数生长期人肝癌HepG2细胞孵育48 h后,提取细胞RNA和总蛋白,分别采用荧光定量PCR法(qRT-PCR)、Western Blot法检测HepG2细胞中CYP3A4 mRNA水平和蛋白水平的表达。体内实验以不同质量浓度的BBR组作为实验组,实验组与空白对照组大鼠分别连续灌胃10 d后,进行大鼠腹股沟动脉插管采血,以底物MDZ作为CYP3A的探针,采用高效液相色谱法(HPLC)测定大鼠血浆中MDZ及其代谢物1′-羟基咪达唑仑(1′-OH-MDZ)的质量浓度,以研究MDZ在大鼠体内的代谢。结果qRT-PCR法分析结果表明,与空白对照组比较,10、20、40μg·mL^(-1) BBR均对HepG2细胞CYP3A4的mRNA表达有显著抑制作用(P<0.05);Western Blot结果表明,与空白对照组比较,0.1、0.5、2.0μg·mL^(-1) BBR对HepG2细胞CYP3A4蛋白的表达有诱导作用(P<0.05),但10、40μg·mL^(-1) BBR对HepG2细胞CYP3A4蛋白的表达有显著抑制作用(P<0.01)。在体内,大鼠口服不同质量浓度药物后MDZ的主要药动学参数为:空白对照组、BBR 50 mg·kg^(-1)组、BBR 100 mg·kg^(-1)组、BBR 200 mg·kg^(-1)组和阳性对照酮康唑75 mg·kg^(-1)组的药时曲线下面积(AUC_((0~t)))分别为(1.25±0.29)、(1.69±0.31)、(2.03±0.44)、(2.12±0.66)、(2.47±0.49)μg·mL^(-1)·h;血药达峰质量浓度(C_(max))分别为(1.13±0.25)、(1.69±0.23)、(1.84±0.29)、(2.12±0.53)、(2.21±0.29)μg·mL^(-1)。各组1′-OH-MDZ的主要药动学参数为:AUC_((0~t))分别为(0.66±0.28)、(0.46±0.19)、(0.42±0.11)、(0.36±0.09)、(0.37±0.10)μg·mL^(-1)·h;C_(max)分别为(0.51±0.16)、(0.44±0.13)、(0.40±0.08)、(0.32±0.07)、(0.33±0.09)μg·mL^(-Objective To study the effects of berberine hydrochloride(BBR)on the mRNA and protein expression of cytochrome P_(450)3A4(CYP3A4)in HepG2 cells and to make sure whether BBR can modify the pharmacokinetic profiles in rats of midazolam(MDZ),a substrate of CYP3A,then to evaluate the correlation in vitro and in vivo.Methods HepG2 cells and control group were treated with different concentrations of BBR for 48 hours.The total proteins and RNA were extracted and the mRNA levels of CYP3A4 in HepG2 cells were assayed with qRT-PCR,and the protein levels of CYP3A4 in HepG2 cells were assayed with Western Blot.The rats were orally administrated with different mass concentration of BBR for 10 days.By using single-pass duodenum perfusion of MDZ,the metabolism of MDZ was investigated in inguinal artery-cannulated rats.The plasma concentrations of MDZ and 1′-OH-MDZ were analyzed by high performance liquid chromatography(HPLC).Results qRT-PCR analysis indicated that the groups administrated with 10,20 and 40μg·mL^(-1) BBR could strongly down-regulate the CYP3A4 mRNA expression in HepG2 cells(P<0.05).Western Blot analysis showed that the groups administrated with 0.1,0.5 and 2μg·mL^(-1) BBR significantly induced the expression of CYP3A4 proteins in HepG2 cells(P<0.05).However,the 10,40μg·mL^(-1) BBR groups could markedly inhibit the expression of CYP3A4 proteins in HepG2 cells(P<0.01).The pharmacokinetic parameters of MDZ after coadministration with BBR were as follows:AUC_((0-t)):(1.25±0.29),(1.69±0.31),(2.03±0.44),(2.12±0.66),(2.47±0.49)μg·mL^(-1)·h.C_(max):(1.13±0.25),(1.69±0.23),(1.84±0.29),(2.12±0.53),(2.21±0.29)μg·mL^(-1).The pharmacokinetic parameters of 1′-hydroxy MDZ after coadministration with BBR were as follows:AUC_((0-t)):(0.66±0.28),(0.46±0.19),(0.42±0.11),(0.36±0.09),(0.37±0.10)μg·mL^(-1)·h;C_(max):(0.51±0.16),(0.44±0.13),(0.40±0.08),(0.32±0.07),(0.33±0.09)μg·mL^(-1).Conclusion In vitro BBR had a dual roles on the expression of CYP3A.Induction was led by low dose of BBR,and i

关 键 词：黄连素(BBR) CYP3A 药代动力学 药物相互作用 

分 类 号：R965[医药卫生—药理学]