質譜條件:以ESI源正離子MRM方式檢測,毛細管溫度320 ℃,毛細管電壓+4 000V,霧化電壓172.375 kPa,幹燥氣流速10 L·min-1,其他質譜分析參數見表2。
2、6 肝微粒體孵育體係孵育體係包括肝微粒體(0、5 g·L-1),NADPH(1 mmol·L-1),CYP特異性探針底物非那西丁(10 μmol·L-1)、甲苯磺丁脲(25 μmol·L-1)、安非他酮(25 μmol·L-1)、美芬妥因(50 μmol·L-1)、右美沙芬(5 μmol·L-1)和咪達唑侖(2、5 μmol·L-1),K2HPO4/KH2PO4緩衝液(0、05 mol·L-1,pH 7、4)補足體係至200 μL。37 ℃水浴預孵育5 min後加入同樣預孵育5 min的NADPH啟動反應,37 ℃水浴孵育30 min,加入200 μL的甲醇-乙腈(1∶1)含有內標鹽酸普奈洛爾100 μg·L-1的溶液終止反應,4 ℃13 000×g離心10 min,取上清液進樣定量分析各底物的代謝產物生成率,測得酶活性。
2、7 數據分析實驗數據以±s表示,對照組和各給藥組之間差異的比較用t檢驗,P1, 2, MA Zeng-chun2*, YI Jian-feng2, WANG Yu-guang2, TAN Hong-ling2,
XIAO Cheng-rong2, LIANG Qian-de2, TANG Xiang-lin2, LI Hua3, SHEN Guo-lin3, GAO Yue2*
(1、 School of Pharmacy, Guangxi Medical University, Nanning , China;
2、 Research Institute of Radiation Medicine, Academy of Military Medical Science, Beijing , China;
3、 Research Institute of Toxicology and Pharmacology, Academy of Military Medical Science, Beijing , China)
[Abstract] To study the effect of Siwu decoction (SWD) compound and its combined administration on hepatic P450 enzymatic activity and mRNA expression in rats、 Rats were orally administered with SWD and water decoction combined with other medicines for two weeks, and then sacrificed、 Their livers were perfused with normal saline to prepare liver micrisomes、 Mixed probe and liver microsome in vitro incubation method were adopted to detect the effect of SWD on hepatic cytochrome P450、 The real-time quantitative polymerase chain reaction (Q-PCR) was used to detect the effect of SWD on the expression of hepatic cytochrome P450、 Compared with the control group, the SWD compound group showed higher CYP1A2 enzymatic activity (P
[Key words] Siwu decoction; liver microsome; cytochrome CYP450 enzyme
doi:10.4268/cjcmm
[責任編輯張寧寧]