异补骨脂素抑制MRP2、MRP3所致的HepG2细胞内胆汁酸蓄积和毒性  

Psoralen induced bile acid accumulation and cytotoxicity by inhibiting MRP2 and MRP3 in HepG2 cells

作　　者：周昆[1,2] 毕亚男[1] 史红[1,2] 

机构地区：[1]天津中医药大学中医药研究院,天津300193 [2]天津中医药大学方剂学教育部重点实验室,天津300193

出　　处：《中国药理学通报》2015年第8期1112-1116,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81202991);国家科技部"重大新药创制"科技重大专项项目(No 2011ZX09201-201-21;No 2014ZX09304307-001-005)

摘　　要：目的观察异补骨脂素体外对Hep G2细胞毒性和细胞内胆汁酸浓度的影响,并考察其对胆汁酸合成转运的影响。方法不同浓度异补骨脂素在Hep G2细胞作用24 h,MTT法检测细胞存活,并检查细胞内胆汁酸浓度。常规TRIzol法提取RNA,real time PCR检测细胞中转运体BSEP、MRP2、MRP3、OATP2、NTCP、OSTα,合成酶CYP7A1、CYP27A1和受体FXR、PXR的m RNA转录水平。结果Hep G2细胞存活率随着异补骨脂素浓度升高而剂量依赖性的降低,异补骨脂素作用24 h的IC50为118.1μmol·L-1。100、400μmol·L-1异补骨脂素可使细胞内胆汁酸浓度明显升高。异补骨脂素在25μmol·L-1时就可使MRP2、MRP3、CYP7A1明显降低,当浓度增大到100μmol·L-1时,OATP2、OSTα、CYP27A1、FXR、PXR也明显升高,但BSEP、NTCP差异无显著性。结论异补骨脂素可引起Hep G2细胞内胆汁酸升高和细胞毒性,其机制可能与抑制MRP2、MRP3有关。Aim To investigate the toxicity of isopsor- Men in HepG2 cells and its effects on bile acid, bile acid synthesis and transport. Methods Cell viability was evaluated by MTT assay and bile acid was deter- mined inside HepG2 cells, with exposure to various isopsoralen for 24h. The mRNA transcription of BSEP, MRP2, MRP3, NTCP, OATP2, OSTα, CYP7A1, CYP27A1, FXR and PXR were assessed by real-time PCR. Results The cell viability was decreased dose- dependently with isopsoralen in HepG2 cells, and IC50 was 118.1 μmol · L-1 exposure to isopsoralen for 24h. Bile acid inside cells significantly increased with 100and 400 μmol · L-1 isopsoralen. Isopsoralen caused the down-regulation of MRP2, MRP3, CYP7A1 mRNA at 25 μmol ·L-1. Beside these, the up-regulation of OATP2, OSTα, CYP27A1, FXR, PXR with 100 μmol · L-1 isopsoralen, but there was no significant change of BSEP and NTCP. Conclusion The results show that isopsoralen induces bile acid accumulation and cytotox- icity which may be associated with the down-regulation of MRP2, MRP3 in HepG2 cells.

关 键 词：异补骨脂素 胆汁酸 细胞毒性 胆汁酸合成 胆汁酸转运 HEPG2细胞 MRP2 MRP3 

分 类 号：R284.1[医药卫生—中药学] R322.47[医药卫生—中医学]