CAJ | 학술논문

目的：研究四物汤及其有效成分对大鼠肝脏 P450酶的影响，从药物代谢酶角度为四物汤的配伍规律提供实验依据。方法大鼠口服灌胃四物汤10 g·kg －1·d －1及其有效成分果糖0.334 g·kg －1·d －1，阿魏酸0.002 g·kg －1·d －1，川芎嗪0.011 g·kg －1·d －1，芍药苷0.022 g·kg －1·d －1，连续给药1周后处死，用生理盐水灌流肝脏，制备肝微粒体，采用混合探针与肝微粒体体外孵育法考察四物汤及其有效成分对大鼠肝脏细胞色素 P450酶的影响。利用实时定量聚合酶链式反应（Q-PCR）检测四物汤及其有效成分对大鼠肝脏P450 mRNA 表达的影响，利用 Western blot 法检测四物汤及其有效成分对大鼠肝脏 CYP2B1蛋白表达的影响。结果肝微粒体孵育结果显示，果糖对 CYP1A2、CYP2B6、CYP2C9、CYP2D6酶活性具有抑制作用，阿魏酸对 CYP2C9、CYP2B6酶活性具有抑制作用，川芎嗪对 CYP1A2、CYP2C9、CYP2B6的酶活性具有抑制作用，芍药苷对 CYP2D6、CYP2B6酶活性具有抑制作用。阿魏酸、果糖、芍药苷组对 CYP2B1mRNA 的表达具有抑制作用，与酶活性水平一致。4种单体成分对CYP2B1蛋白表达具有抑制作用。结论阿魏酸、果糖、芍药苷组在 CYP450酶活性、mRNA 基因转录水平及蛋白表达水平对 CYP2B1均有抑制作用。
목적：연구사물탕급기유효성분대대서간장 P450매적영향，종약물대사매각도위사물탕적배오규률제공실험의거。방법대서구복관위사물탕10 g·kg －1·d －1급기유효성분과당0.334 g·kg －1·d －1，아위산0.002 g·kg －1·d －1，천궁진0.011 g·kg －1·d －1，작약감0.022 g·kg －1·d －1，련속급약1주후처사，용생리염수관류간장，제비간미립체，채용혼합탐침여간미립체체외부육법고찰사물탕급기유효성분대대서간장세포색소 P450매적영향。이용실시정량취합매련식반응（Q-PCR）검측사물탕급기유효성분대대서간장P450 mRNA 표체적영향，이용 Western blot 법검측사물탕급기유효성분대대서간장 CYP2B1단백표체적영향。결과간미립체부육결과현시，과당대 CYP1A2、CYP2B6、CYP2C9、CYP2D6매활성구유억제작용，아위산대 CYP2C9、CYP2B6매활성구유억제작용，천궁진대 CYP1A2、CYP2C9、CYP2B6적매활성구유억제작용，작약감대 CYP2D6、CYP2B6매활성구유억제작용。아위산、과당、작약감조대 CYP2B1mRNA 적표체구유억제작용，여매활성수평일치。4충단체성분대CYP2B1단백표체구유억제작용。결론아위산、과당、작약감조재 CYP450매활성、mRNA 기인전록수평급단백표체수평대 CYP2B1균유억제작용。
Aim To study the influence of Si-Wu De-coction (SWD ) and its active components on cyto-chrome P450 activity and mRNA expression in rats in order to provide an experimental basis for compatibility of SWD.Methods SWD and its active components were intragastrically administrated for seven days,the doses of SWD was 10 g · kg -1 · d -1 ,the doses of fructose,ferulic acid,ligustrazine,peoniflorin were 0.334,0.002,0.011 and 0.022 g·kg -1 ·d -1 ,re-spectively.After administration for seven days,rats were executed,and liver microsomes were prepared. The effects of SWD and its active components on cyto-chrome P450 in rats were investigated by hybrid probe and liver microsomes incubation method.The level of mRNA expression in liver was detected by real-time quantitative polymerase chain reaction using specific target primers for CYP450 genes.The level of protein expression of CYP2B1 was detected by Western blot. Results Compared with the control group,fructose significantly decreased the activity of CYP1A2, CYP2B6,CYP2C9,CYP2D6;ferulic acid significantly decreased the activity of CYP2C9,CYP2B6;ligus-trazine significantly decreased the activity of CYP1A2, CYP2C9,CYP2B6;peoniflorin significantly decreased the activity of CYP2D6,CYP2B6;fructose,ferulic acid,peoniflorin inhibited the mRNA expression of CYP2B1;fructose,ferulic acid,ligustrazine and peon-iflorin also inhibit the protein expression of CYP2B1. Conclusion Fructose,ferulic acid,peoniflorin inhib-it the activity of CYP2B1,decrease the expression lev-els of mRNA and protein of CYP2B1.