| 米琼宇,吉金子,邰 婷,谷彤彤,周 缓.黄芪甲苷IV调节MRP3的表达及功能研究[J].,2019,19(23):4407-4410 |
| 黄芪甲苷IV调节MRP3的表达及功能研究 |
| The Molecular Mechanism Underlying the Induction of MRP3 Expression and Function by Astragaloside IV |
| 投稿时间：2019-06-21 修订日期：2019-07-17 |
| DOI：10.13241/j.cnki.pmb.2019.23.002 |
| 中文关键词: 黄芪甲苷IV 多药耐药相关蛋白-3 核因子E2相关因子2 药物-药物相互作用 |
| 英文关键词: Astragaloside IV MRP3 Nrf2 Drug-drug interactions |
| 基金项目:国家自然科学基金青年科学项目（81503144）；南京市医学科技发展资金项目（QRX17069） |
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| 中文摘要: |
| 摘要 目的：研究黄芪甲苷IV（AS-IV）对人肝癌细胞HepG2中多药耐药相关蛋白-3（MRP3）的调节作用，为AS-IV可能引发的药物相互作用阐明机制。方法：培养HepG2细胞，用不同浓度AS-IV进行干预，Western blotting方法检测细胞中MRP3的蛋白水平，流式细胞术检测胞内5-羧基荧光素（5-CF）的荧光强度，反映MRP的功能；给予MRP3 siRNA干扰，观察AS-IV诱导的MRP3蛋白表达是否增加其外排转运功能；提取胞核、总蛋白，检测胞核核因子E2相关因子2（Nrf2）、总Nrf2蛋白水平；给予Nrf2 siRNA干扰，观察AS-IV诱导的MRP3蛋白表达是否依赖于Nrf2。结果：200 μmol/L AS-IV作用48 h后，细胞MRP3的蛋白水平显著增加（P<0.05），胞内5-CF的平均荧光强度（MFI）明显低于对照组（P<0.05），与单独给予AS-IV组相比，给予MRP3 siRNA干扰显著增加了胞内5-CF的MFI（P<0.05）。AS-IV增加了胞核内Nrf2的蛋白水平（P<0.01），亦增加了总Nrf2的蛋白表达（P<0.01）。给予Nrf2 siRNA干扰后，AS-IV诱导上调的MRP3被显著抑制（P<0.05）。结论： |
| 英文摘要: |
| ABSTRACT Objective: To investigate the regulation of astragaloside IV (AS-IV) on multidrug resistance associated protein 3 (MRP3) in HepG2 cells. Results would provide information for AS-IV-induced drug-drug interactions (DDIs) during clinical application. Methods: Cultured HepG2 cells were administered with AS-IV at different concentrations. Protein levels of MRP3 were determined by Western blotting. Mean fluorescence intensity (MFI) of 5-carboxyfluorescein (5-CF) was measured by flow cytometry to determine MRP function. MRP3 siRNA knock-down was used to determine whether the MRP3 protein induced by AS-IV possessed an enhanced efflux transport. Nuclear and total protein were extracted, and both nuclear and total nuclear factor erythroid 2-related factor 2 (Nrf2) were determined by Western blotting. Nrf2 siRNA knock-down was used to determine whether the induction of MRP3 protein by AS-IV was Nrf2 dependent. Results: 48 h treatment of AS-IV at the dose of 200 μmol/L caused a significant increase in MRP3 protein levels(P<0.05). The MFI value of 5-CF retained in cells treated with AS-IV was lower than that in vehicle-treated cells(P<0.05). Pretreatment with MRP3 siRNA led to an increased MFI, when compared with AS-IV alone(P<0.05). Furthermore, AS-IV remarkably increased both nuclear (P<0.01) and total Nrf2(P<0.01) protein levels in cells. AS-IV-induced increase in MRP3 protein expression was attenuated by pretreatment with Nrf2 siRNA(P<0.05). Conclusion: AS-IV induces MRP3 protein expression and enhances MRP3 efflux pumping activity. Activation of Nrf2 plays an important role in induction of MRP3 protein levels by AS-IV. These results suggested that potentially DDIs likely occurred when AS-IV was used concomitantly with other drugs that are substrates of MRP3. |
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