目的 使用原代培養(yǎng)的人肝細胞研究人參皂苷Rb₁和Rg₁、丹參素鈉、冰片對細胞色素P450酶（CYP450）的誘導(dǎo)作用。方法 分別將3批次的冷凍原代人肝貼壁細胞進行接種培養(yǎng)，使用人參皂苷Rb₁和Rg₁、丹參素鈉、冰片（30 μmol/L）對CYP1A2、CYP2B6和CYP3A4進行誘導(dǎo)，實時熒光定量PCR（qRT-PCR）法測定CYP450酶mRNA表達水平，評價4種單體成分對P450酶的誘導(dǎo)作用。陽性對照為20 μmol/L利福平（CYP3A4誘導(dǎo)劑）、50 μmol/L奧美拉唑鈉（CYP1A2誘導(dǎo)劑）和1 mmol/L苯巴比妥鈉（CYP2B6誘導(dǎo)劑）；陰性對照組為10 μmol/L紅霉素。冰片（30 μmol/L）與利福平聯(lián)合給藥，觀察冰片對利福平的CYP1A2、CYP2B6和CYP3A4誘導(dǎo)作用的影響。結(jié)果 3批次人肝細胞的誘導(dǎo)結(jié)果顯示，陽性誘導(dǎo)劑奧美拉唑、苯巴比妥、利福平分別顯著誘導(dǎo)CYP1A2、CYP2B6、CYP3A4的表達，陰性對照紅霉素對CYP1A2、CYP2B6及CYP3A4的mRNA表達水平未見明顯影響，人參皂苷Rb₁和Rg₁、丹參素鈉、冰片在30 μmol/L濃度給藥時，對3批次人肝細胞的CYP1A2、CYP2B6和CYP3A4 mRNA表達水平未見明顯影響；30 μmol/L冰片與利福平聯(lián)合給藥，與利福平單獨給藥比較，3批次肝細胞的CYP2B6、CYP3A4 mRNA表達水平均減少，對CYP3A4的mRNA表達水平影響尤為顯著。結(jié)論 人參皂苷Rb₁和Rg₁、丹參素鈉、冰片在30 μmol/L濃度給藥時，對3批次人肝細胞的CYP1A2、CYP2B6和CYP3A4均沒有誘導(dǎo)作用。冰片與利福平聯(lián)合用藥時，可以阻斷利福平對CYP2B6和CYP3A4的誘導(dǎo)作用，對CYP3A4的影響尤為顯著。

Objective In this study, human primary cultured hepatocytes were used to study the induction of P450 enzyme by ginsenoside Rb₁, Rg₁, Danshensu Sodium and Borneol. Methods The CYP1A2, CYP2B6 and CYP3A4 were induced by different inducers, and the real-time RT-PCR method was used to determine the mRNA expression level. The results of CYP450 mRNA expression level was used to evaluate the induction of P450 enzyme. The positive control group was 20 μmol/L rifampicin (CYP3A4 inducer), 50 μmol/L omeprazole sodium (CYP1A2 inducer) and 1 mmol/L phenobarbital sodium (CYP2B6 inducer); the negative control group was 10 μmol/L erythromycin. Borneol (30 μmol/L) combined with rifampicin was used to observe the effect of Borneol on the induction of CYP1A2, CYP2B6 and CYP3A4 of rifampicin. Results The results of three donors of induction showed that the expression of CYP1A2, CYP2B6 and CYP3A4 was significantly induced by omeprazole, phenobarbital sodium and rifampicin respectively, and the corresponding mRNA expression levels were significantly increased, all > 4; negative control erythromycin which levels of CYP1A2, CYP2B6 and CYP3A4 mRNA were not significantly change. The expression levels of CYP1A2, CYP2B6 and CYP3A4 mRNA were not significantly affected by ginsenoside Rb₁, Rg₁, danshensu sodium and borneol at the concentrations of 30 μmol/L. Comparedwithrifampinalone,when borneol (30 μmol/L) was combined with rifampicin (20 μmol/L), the expression of CYP2B6 and CYP3A4 mRNA in the three batches of hepatocytes decreased, and the effect on the expression of CYP3A4 mRNA was particularly significant. No significant effect of CYP1A2 mRNA expression levels was seen. Conclusion The expression levels of CYP1A2, CYP2B6 and CYP3A4 mRNA were not significantly affected by Rb₁, Rg₁, danshensu sodium and borneol at the concentrations of 30 μmol/L. When borneol (30 μmol/L) was combined with rifampicin (20 μmol/L), there was no effect on CYP1A2 in the three batches of hepatocytes, but the induction effect of rifampicin on CYP2B6 and CYP3A4 in three batches of hepatocytes were reduced while the decrease of CYP3A4 is particularly evident.

R969.1

國家“重大新藥創(chuàng)制”科技重大專項（2012ZX09304002）；國家自然科學(xué)基金青年科學(xué)基金項目（81503154）