Article

Journal of Huazhong University of Science and Technology [Medical Sciences]

, Volume 33, Issue 1, pp 75-80

Copper ions stimulate the proliferation of hepatic stellate cells via oxygen stress in vitro

  • San-qing Xu徐三清Affiliated withDepartment of Pediatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology
  • , Hui-yun Zhu朱慧云Affiliated withDepartment of Pediatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology
  • , Jian-guo Lin林剑国Affiliated withDepartment of Internal Medicine, School of Medicine, Saint Louis University
  • , Tang-feng Su苏堂枫Affiliated withDepartment of Pediatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology
  • , Yan Liu刘 艳Affiliated withDepartment of Pediatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology
  • , Xiao-ping Luo罗小平Affiliated withDepartment of Pediatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology Email author 

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Summary

This study examined the effect of copper ions on the proliferation of hepatic stellate cells (HSCs) and the role of oxidative stress in this process in order to gain insight into the mechanism of hepatic fibrosis in Wilson’s disease. LX-2 cells, a cell line of human HSCs, were cultured in vitro and treated with different agents including copper sulfate, N-acetyl cysteine (NAC) and buthionine sulfoximine (BSO) for different time. The proliferation of LX-2 cells was measured by non-radioactive cell proliferation assay. Real-time PCR and Western blotting were used to detect the mRNA and protein expression of platelet-derived growth factor receptor β subunit (PDGFβR), ELISA to determine the level of glutathione (GSH) and oxidized glutathione (GSSG), dichlorofluorescein assay to measure the level of reactive oxygen species (ROS), and lipid hydroperoxide assay to quantify the level of lipid peroxide (LPO). The results showed that copper sulfate over a certain concentration range could promote the proliferation of LX-2 cells in a time- and dose-dependent manner. The effect was most manifest when LX-2 cells were treated with copper sulfate at a concentration of 100 μmol/L for 24 h. Additionally, copper sulfate could dose-dependently increase the levels of ROS and LPO, and decrease the ratio of GSH/GSSG in LX-2 cells. The copper-induced increase in mRNA and protein expression of PDGFβR was significantly inhibited in LX-2 cells pre-treated with NAC, a precursor of GSH, and this phenomenon could be reversed by the intervention of BSO, an inhibitor of NAC. It was concluded that copper ions may directly stimulate the proliferation of HSCs via oxidative stress. Anti-oxidative stress therapies may help suppress the copper-induced activation and proliferation of HSCs.

Key words

copper ions hepatic stellate cells oxidative stress hepatic fibrosis