Astragali radix and its main bioactive compounds activate the Nrf2-mediated signaling pathway to induce P-glycoprotein and breast cancer resistance protein
Astragali radix and its main bioactive compounds activate the Nrf2-mediated signaling pathway to induce P-glycoprotein and breast cancer resistance protein
复制标题
黄芪及其主要生物活性化合物激活Nrf2介导的信号通路诱导P-糖蛋白和乳腺癌耐药蛋白
DOI：
10.1016/j.jep.2018.09.026
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发表时间：
2019-01-10
影响因子：
5.4
通讯作者：
Wu, Jinjun
中科院分区：
文献类型：
--
作者：
Lou, Yanmei;Guo, Zhenzhen;Wu, Jinjun
民族药理学相关性：黄芪是一种著名的中草药，广泛与许多其他药物联合用药治疗疾病。潜在的草药-药物相互作用（HDI）可能发生在联合治疗期间。 P-糖蛋白（P-gp）和乳腺癌抗性蛋白（BCRP）是介导HDI产生的关键靶标。我们之前观察到HQ及其三种主要生物活性化合物，包括黄芪甲苷IV（AS-IV）、毛蕊异甙（CS）和芒柄花素（FMNT），能够在体外显着诱导HepG2细胞中P-gp和BCRP的表达。然而，它们对 P-gp 和 BCRP 功能的调节仍然未知。它们对这两种蛋白体内表达的影响尚不清楚；研究目的：本研究旨在探讨HQ、AS-IV、CS和FMNT对体内P-gp和BCRP的影响及其具体调控机制。还研究了HQ及这些化合物对P-gp和BCRP功能的影响。材料和方法：野生型C57BL/6小鼠和核因子E2相关因子2敲除(Nrf2(-/-))C57BL/6小鼠口服HQ、AS-IV、CS或FMNT。采用Western blot和免疫组化方法检测小鼠肝脏中P-gp和BCRP的蛋白水平。通过使用实时PCR测量mRNA水平。通过在稳定转染的 HepG2-C8 细胞中使用报告基因测定来研究药物对抗氧化反应元件 (ARE)-荧光素活性的激活。采用流式细胞仪和典型探针检测HepG2细胞中P-gp和BCRP的外排活性。结果：HQ、AS-IV、CS和FMNT显着上调野生型小鼠肝脏中P-gp和BCRP的表达。 Nrf2(-/-) 小鼠中的诱导作用显着逆转。 HQ 和这些化合物显着增加了野生型小鼠中 Nrf2 的表达。 HQ和这些化合物还显着增强了ARE-荧光素活性并促进了细胞中Nrf2的核转位。此外，HQ和这些化合物显着增强了P-gp和BCRP的外排活性，并增加了细胞内ATP水平。结论：我们的结果证明HQ及其主要生物活性化合物可以通过激活Nrf2介导的信号通路来诱导P-gp和BCRP的表达。 HQ 和这些化合物还显着增强了 P-gp 和 BCRP 的外排活性，细胞内 ATP 水平的增加可能与 P-gp 和 BCRP 功能的增强有关。这些结果表明，当 HQ 与作为 P-gp 和 BCRP 底物的其他药物同时使用时，可能会发生潜在的 HDI。
Ethnopharmacological relevance: Astragali radix (Huang Qi, HQ), a well-known Chinese herbal medicine, is widely coadministered with many other drugs for treating diseases. The potential herb-drug interactions (HDIs) possibly occur during the combination therapy. P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP) are the crucial targets that mediate the production of HDIs. We previously observed that HQ and its three main bioactive compounds, including Astragaloside IV (AS-IV), calycosin (CS) and formononetin (FMNT), could significantly induce the expression of P-gp and BCRP in HepG2 cells in vitro. However, their modulations on the function of P-gp and BCRP remain unknown; their impact on these two proteins expression in vivo is not clear; the exact regulatory mechanism has also not yet been explored.Aim of the study: This study aimed to investigate the impact of HQ, AS-IV, CS and FMNT on P-gp and BCRP in vivo, and the exact regulatory mechanism involved. The effects of HQ and these compounds on the function of P-gp and BCRP were also studied.Materials and methods: Wild-type C57BL/6 mice and nuclear factor E2-related factor-2 knockout (Nrf2(-/-)) C57BL/6 mice were orally treated with HQ, AS-IV, CS or FMNT. The protein levels of P-gp and BCRP in the liver of mice were measured by using Western blot and immunohistochemistry. The mRNA levels were measured by using real-time PCR. The activation of the drugs on the antioxidant response element (ARE)-luciferin activity was studied by using reporter assay in a stably transfected HepG2-C8 cells. The efflux activity of P-gp and BCRP in HepG2 cells were tested by using flow cytometer with typical probes.Results: HQ, AS-IV, CS and FMNT significantly upregulated the P-gp and BCRP expression in the liver of wild type mice. The induction was significantly reversed in the Nrf2(-/-) mice. HQ and these compounds significantly increased the Nrf2 expression in wild-type mice. HQ and these compounds also markedly enhanced the ARE-luciferin activity and promoted the nuclear translocation of Nrf2 in cells. Besides, HQ and these compounds significantly enhanced the efflux activity of P-gp and BCRP, and increased the intracellular ATP levels.Conclusions: Our results proved that HQ and its main bioactive compounds could induce the P-gp and BCRP expression through the activation of the Nrf2-mediated signaling pathway. HQ and these compounds also significantly enhanced the efflux activity of P-gp and BCRP, and the increased intracellular ATP levels were likely involved in the increased P-gp and BCRP function. These results suggested that potentially HDIs likely occurred when HQ was used concomitantly with other drugs that are substrates of P-gp and BCRP.