目的 考察烏頭堿配伍人參皂苷Rb₁、甘草苷后對HepG2藥物代謝酶（Cytochrome P450，CYP450）中3A4亞型的報告基因熒光活性、mRNA轉錄及蛋白翻譯水平的影響。方法 將pGLuc-CYP3A4報告基因質粒與pcDNA3.1-hPXR表達質粒共轉染HepG2細胞，檢測烏頭類生物堿、人參皂苷和甘草的單體成分對CYP3A4的激活效應；并利用實時熒光定量PCR（qRT-PCR）及Western blotting技術檢測人參皂苷Rb₁、甘草苷與烏頭堿對CYP3A4 mRNA及蛋白水平的影響。結果 報告基因模型檢測結果顯示，與對照組比較，烏頭堿、新烏頭堿、次烏頭堿和乙酰烏頭堿能下調CYP3A4報告基因熒光強度（P<0.05），其中烏頭堿下調能力最強，人參皂苷Rb₁、Rc、Re、Rg1以及甘草苷、異甘草苷、甘草素和甘草酸均能上調報告基因的熒光強度（P<0.05、0.01），其中人參皂苷Rb₁和甘草苷上調能力最明顯；同時烏頭堿能下調CYP3A4 mRNA與蛋白表達水平（P<0.05、0.01），人參皂苷Rb₁和甘草苷能逆轉烏頭堿下調CYP3A4的能力（P<0.05、0.01）。結論 人參皂苷Rb₁、甘草苷與烏頭堿配伍后可上調CYP3A4的表達，減少烏頭堿在體內蓄積時間，起到減毒的作用。

Objective To investigate the effects of aconitine combined with ginsenoside Rb₁ and liquiritin on the fluorescence activity, transcription level and protein translation level of 3A4 subtype of hepatocyte cytochrome P450 (CYP450) in HepG2. Methods The pGLuc-CYP3A4 reporter gene plasmid and pcDNA3.1-hPXR expression plasmid were co-transfected into HepG2 cells to detect the activation effect of aconitines, ginsenoside and components of Glycyrrhiza uralensis on CYP3A4. The expression of ginsenoside Rb₁, liquiritin and aconitine was detected by qRT-PCR and Western blotting. Resluts The results of reporter gene model test showed that aconitine, mesaconitine, hypaconitine and aconitine 3-acetate could down-regulate the fluorescence activity of CYP3A4 reporter gene (P<0.05). Aconitine had the strongest down-regulation ability. Ginsenoside Rb₁, Rc, Re, Rg1 and liquiritin, iso liquiritin, liquiritigenin and glycyrrhiza acid could up-regulate the fluorescence activity of reporter gene (P<0.05 and 0.01), among which ginsenoside Rb₁ and liquiritin were more up-regulation. At the same time, aconitine can down-regulate the expression of CYP3A4 and ginsenoside Rb₁ and liquiritin can reverse the ability of aconitine to down-regulate CYP3A4 (P<0.05, 0.01). Conclusion Ginsenoside Rb₁, liquiritin and aconitine can up-regulate the expression of CYP3A4, reduce the accumulation time of aconitine in vivo and play a role in reducing toxicity.

R285.5

國家重點研發(fā)計劃資助（2018YFC1708204）