Up-regulation of Nrf2 pathway and intervention of intracellular redox homeostasis in shikonin-induced A549 cell apoptosis
10.3969/j.issn.1001-1978.2014.10.024
- VernacularTitle:紫草素通过上调 Nrf2途径及干预胞内氧化还原平衡稳态诱导A549细胞凋亡
- Author:
Chen XIE
;
Hanying CHEN
;
Jing ZHONG
;
Xiaoqin WANG
;
Bo ZHANG
- Publication Type:Journal Article
- Keywords:
shikonin;
A549 cell;
apoptosis;
ROS;
Nrf2 pathway;
redox homeostasis
- From:
Chinese Pharmacological Bulletin
2014;(10):1445-1451
- CountryChina
- Language:Chinese
-
Abstract:
Aim To investigate the roles of intracellu-lar reactive oxygen species ( ROS ) and Nrf2 pathway in shikonin-induced A549 cell apoptosis. Methods The cytotoxicity was analyzed by MTT assay. The ap-optosis of A549 cells was analyzed by both cellular morphological and biochemical methods. The relative changes of the redox marks ( ROS/GSH) were studied by fluorescence assay in the shikonin-treated A549 cells in accompany with the changes of the intracellular redox homeostasis by GSH/GSSG ratio. ROS inhibitor was also employed in the treatment to find the role of ROS in shikonin-induced A549 cell apoptosis. Real-time PCR analysis and ELISA assay were performed as well to determine the role of Nrf2 pathway in the shiko-nin-induced A549 cell apoptosis. Results The IC50 of shikonin on A549 cells was 3. 2 mg·L-1 . The cellu-lar redox homeostasis shifted toward oxidation signifi-cantly in shikonin treatment in a time-dependent man-ner. The expression of the Nrf2 pathway related genes was up-regulated by shikonin ( 3 . 2 mg · L-1 , 8 h ) . The expression of the anti-apoptotic genes was down-regulated , and proapoptotic genes were up-regulated by shikonin (3. 2 mg·L-1, 24h). Futhermore, the inhi-bition of intracellular ROS alleviated the cytotoxicity of shikonin in A549 cells. Conclusion The critical role of shikonin-induced redox imblance in A549 cell, coped with the secondary produced ROS and Nrf2 path-way antioxidants, result in A549 cell apoptosis.
