目的 考察烏頭堿配伍人參皂苷Rb₁、甘草苷后對(duì)HepG2藥物代謝酶（Cytochrome P450，CYP450）中3A4亞型的報(bào)告基因熒光活性、mRNA轉(zhuǎn)錄及蛋白翻譯水平的影響。方法 將pGLuc-CYP3A4報(bào)告基因質(zhì)粒與pcDNA3.1-hPXR表達(dá)質(zhì)粒共轉(zhuǎn)染HepG2細(xì)胞，檢測(cè)烏頭類生物堿、人參皂苷和甘草的單體成分對(duì)CYP3A4的激活效應(yīng)；并利用實(shí)時(shí)熒光定量PCR（qRT-PCR）及Western blotting技術(shù)檢測(cè)人參皂苷Rb₁、甘草苷與烏頭堿對(duì)CYP3A4 mRNA及蛋白水平的影響。結(jié)果 報(bào)告基因模型檢測(cè)結(jié)果顯示，與對(duì)照組比較，烏頭堿、新烏頭堿、次烏頭堿和乙酰烏頭堿能下調(diào)CYP3A4報(bào)告基因熒光強(qiáng)度（P<0.05），其中烏頭堿下調(diào)能力最強(qiáng)，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(diào)報(bào)告基因的熒光強(qiáng)度（P<0.05、0.01），其中人參皂苷Rb₁和甘草苷上調(diào)能力最明顯；同時(shí)烏頭堿能下調(diào)CYP3A4 mRNA與蛋白表達(dá)水平（P<0.05、0.01），人參皂苷Rb₁和甘草苷能逆轉(zhuǎn)烏頭堿下調(diào)CYP3A4的能力（P<0.05、0.01）。結(jié)論 人參皂苷Rb₁、甘草苷與烏頭堿配伍后可上調(diào)CYP3A4的表達(dá)，減少烏頭堿在體內(nèi)蓄積時(shí)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(guó)家重點(diǎn)研發(fā)計(jì)劃資助（2018YFC1708204）