基于细胞色素P450 2C9的基因多态性研究丹参组分对氟伐他汀代谢影响及其机制  

Study on the effect of the components of salviae miltiorrhizae on the metabolism of fluvastatin and its mechanism based on the polymorphism of cytochrome P450 2C9

作　　者：马桂臣 段译斐 张红[1] 靳高凤 刘名义[1] 熊玉卿[1] 

机构地区：[1]南昌大学临床药理研究所,南昌330006

出　　处：《中国临床药理学杂志》2017年第21期2171-2174,共4页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81260508);江西省"赣鄱英才555工程"人才资助计划基金资助项目(18000046)

摘　　要：目的探讨丹参的6种组分(丹参素、原儿茶酸、丹酚酸B、隐丹参酮、二氢丹参酮Ⅰ、丹参酮ⅡA)对人肝微粒体代谢氟伐他汀的影响,并基于细胞色素P450 2C9(CYP2C9)*1,*2,*3基因多态性,研究隐丹参酮和二氢丹参酮Ⅰ对氟伐他汀代谢的影响。方法构建肝微粒/不同基因型重组酶孵育体系。用HPLC检测氟伐他汀浓度,通过软件求算酶动力学参数以及半数抑制浓度(IC_(50))和抑制常数(Ki)值,比较不同突变位点CYP2C9代谢氟伐他汀的能力及丹参组分对其的影响。结果人肝微粒体实验显示:在隐丹参酮和二氢丹参酮Ⅰ10μmol·L^(-1)时,其对氟伐他汀代谢的剩余酶活性分别为30.24%和14.52%,显示出了明显的抑制作用,然而其他丹参组分则没有显示出抑制作用。CYP2C9重组酶实验:CYP2C9*1,*2,*3代谢氟伐他汀的米氏常数(Km)分别为(1.56±0.21),(1.79±0.25)和(2.32±0.43)μmol·L^(-1),最大反应速率(Vmax)分别为(41.75±1.67),(26.55±1.08)和(12.85±0.79)pmol·min^(-1)·mg^(-1),内在清除率(CLint)分别为26.71,14.86和5.53μL·min^(-1)·mg^(-1),显示CYP2C9*1的代谢活性强于*2,*3;隐丹参酮和二氢丹参酮Ⅰ对CYP2C9*1,*2,*3代谢氟伐他汀的IC_(50)值分别是(1.74±0.41),(2.64±0.81),(3.17±0.32)μmol·L^(-1)和(0.19±0.09),(0.56±0.16),(1.52±0.34)μmol·L^(-1),CYP2C9*1的IC_(50)和Ki值与*3比较,差异均有统计学意义(P<0.05,P<0.01)。结论 CYP2C9*2(430 C>T)和*3(1075A>C)突变可影响其代谢活性。隐丹参酮和二氢丹参酮Ⅰ能明显抑制CYP2C9代谢氟伐他汀,且其对不同突变CYP2C9代谢氟伐他汀的抑制能力是有差别的。Objective To identify the effect of six components of salviae miltiorrhizae on the metabolism of fluvastatin in the human liver microsomes（ HLMs） and to further explore the effect of cryptotanshinone and dihydrotanshinone I on the metabolism of fluvastatin based on the polymorphism（ ＊ 1,＊ 2,＊ 3） of cytochrome P450 2 C9（ CYP2 C9）.Methods Establishing the incubation system of HLMs and recombinase and determining the concentration of fluvastatin by HPLC. To calculate the enzyme kinetic parameters, half maximal inhibitory concentration（ IC50） and inhibition constant（ Ki） so as to compare the impact of different CYP2 C9 mutation site on the metabolism of fluvastatin and the influence of components of salviae miltiorrhizae on the metabolismprocess. Results It was shown that cryptotanshinone and dihydrotanshinone Ⅰ exerted significant inhibition on the metabolism of fluvastatin and the residual activity were 30. 24%,14. 52% respectively when the concentrations were 10μmol·L^-1 in the HLMs. However,other exhibited no inhibitory effect. The michaelis constant（ Km）,maximum velocity（ Vmax） and intrinsic clearance（ CLint） of CYP2 C9 ＊ 1,＊ 2,＊ 3 recombinase-catalyzed fluvastatin was（ 1. 56 ± 0. 21）,（ 1. 79 ± 0. 25）,（ 2. 32 ± 0. 43） μmol·L^-1,（ 41. 75 ± 1. 67）,（ 26. 55 ± 1. 08）,（ 12. 85 ± 0. 79）pmol·min^-1·mg^-1 and 26. 71,14. 86,5. 53 μL·min^-1·mg^-1 respectively. It was indicated that the metabolic activity of CYP2 C9＊ 1 was stronger than ＊ 2,＊ 3. The IC50 value of CYP2 C9＊ 1,＊ 2,＊ 3 recombinase-catalyzed fluvastatin by cryptotanshinone and dihydrotanshinone I was（ 1. 74 ± 0. 41）,（ 2. 64 ± 0. 81）,（ 3. 17 ± 0. 32）μmol·L^-1 and（ 0. 19 ± 0. 09）,（ 0. 56 ± 0. 16）,（ 1. 52 ± 0. 34） μmol·L^-1. The IC50 and Kivalue of CYP2 C9＊ 1 was less than ＊ 2 and ＊ 3 but there was statistical significance between the ＊ 1 and ＊ 3 with respect to the difference of IC50 and Ki. Conclusion The mutation site of 4

关 键 词：丹参组分 氟伐他汀 细胞色素P450 2C9 基因多态性 代谢 

分 类 号：R972[医药卫生—药品]