Tanshinone IIA prevents rifampicin-induced liver injury by regulating BSEP/NTCP expression via epigenetic activation of NRF2
Tanshinone IIA prevents rifampicin-induced liver injury by regulating BSEP/NTCP expression via epigenetic activation of NRF2
复制标题
丹参酮 IIA 通过 NRF2 表观遗传激活调节 BSEP/NTCP 表达，预防利福平诱导的肝损伤
DOI：
10.1111/liv.14262
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发表时间：
2019-10-07
影响因子：
6.7
通讯作者：
Wang, Ling
中科院分区：
文献类型：
--
作者：
Yang, Yujie;Liu, Lei;Wang, Ling
背景与目的利福平（RFP）诱导的胆汁淤积性肝损伤以胆汁酸（BA）转运受损为特征。胆盐外排泵（BSEP）和Na +/牛磺胆酸协同转运蛋白（NTCP）是BA的主要转运蛋白。然而，人们对这些转运蛋白的机制知之甚少。方法从核因子红细胞2相关因子2（NRF2）-BSEP/NTCP信号通路的调控机制出发，观察丹参酮IIA（TAN IIA）对RFP诱导的肝损伤的保护作用。研究了TAN IIA对NRF 2的表观遗传诱导作用以及对BSEP和NTCP转录激活和NRF 2 DNA结合能力的影响。结果TAN Ⅱ A可明显诱导肝细胞BSEP和NTCP的表达。NRF2敲除废除了诱导。我们在人BSEP启动子上发现了两个NRF2结合位点，称为肌肉腱膜纤维肉瘤识别元件（MARE），在NTCP启动子上发现了一个MARE。人BSEP和NTCP启动子荧光素酶报告基因质粒由NRF 2刺激。预测的MARE的突变消除了NRF2的转录激活。TAN IIA诱导TET2的表达，介导NRF2的去甲基化，从而促进NRF2 DNA结合在BSEP和NTCP启动子上并激活其转录。最后，在体内，Nrf2在RFP诱导的肝损伤（Nrf2-/-小鼠中更严重的肝损伤）中发挥了重要作用，TAN IIA阻止了it.Conclusions这些结果表明，NRF2调节靶转运蛋白BSEP和NTCP，依赖于TET2的DNA去甲基化。TAN IIA对NRF2的药理学激活可能对RFP诱导的肝损伤有益。
Background & Aims Rifampicin (RFP)-induced cholestatic liver injury is characterized by impaired hepatic bile acid (BA) transport. Bile salt efflux pump (BSEP) and Na+/taurocholate cotransporter (NTCP) are the major BA transporters. However, little is known about the mechanisms underlying these transporters. Methods The role of tanshinone IIA (TAN IIA) in preventing RFP-induced liver injury was evaluated in vitro and in vivo, based on the regulatory mechanism of nuclear factor erythroid 2-related factor 2 (NRF2)-BSEP/NTCP signalling. The epigenetic induction of NRF2 by TAN IIA was investigated as well as the influence on BSEP and NTCP transcriptional activation and NRF2 DNA-binding ability. Results TAN IIA strongly induced BSEP and NTCP expression in hepatocytes. NRF2 knockdown abrogated the induction. We found two NRF2 binding sites on the human BSEP promoter, called musculoaponeurotic fibrosarcoma recognition elements (MAREs), and one MARE on the NTCP promoter. Human BSEP and NTCP promoter luciferase reporter gene plasmids were stimulated by NRF2. Mutations of the predicted MAREs abolished NRF2 transcriptional activation. TAN IIA induced the expression of ten-eleven translocation 2 (TET2) to mediate the demethylation of NRF2, which promoted NRF2 DNA-binding on the BSEP and NTCP promoters and their transcriptional activation. Finally, in vivo, Nrf2 played an important role in RFP-induced liver injury (more serious liver injury in Nrf2-/- mice), and TAN IIA prevented it. Conclusions These results indicate that NRF2 regulates the target transporters BSEP and NTCP, depending on the DNA demethylation by TET2. Pharmacological activation of NRF2 by TAN IIA may be beneficial for RFP-induced liver injury.