Involvement of Mitochondrial Pathway in Triptolide-Induced Cytotoxicity in Human Normal Liver L-02 Cells

抄録

Triptolide, a purified diterpenoid triepoxide compound derived from a traditional Chinese medicine, Tripterygium wilfordii HOOK. f (TWHf), has been used in the treatment of autoimmune and inflammatory diseases. However, the toxicity of triptolide limits its application to a great extent. In the present study, we treated human normal liver L-02 cells (L-02 cells) with triptolide in vitro and investigated its toxic effects. The cytotoxicity was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay for cellular viability and by flow cytometry and Hoechst 33258 staining for apoptosis. The mitochondrial membrane potential (Δψm) was evaluated by flow cytometry with JC-1 as probe. After treatment with triptolide, a decrease in the viability of L-02 cells and increase in apoptosis were observed. Triptolide-induced apoptosis was accompanied by loss of mitochondrial membrane potential and release of cytochrome c (cyt-c) from the mitochondria to the cytosol and down-regulation of anti-apoptotic protein Bcl-2 levels with concurrent up-regulation in pro-apoptotic protein Bax levels and tumor suppressor protein p53 levels. Triptolide-increased activity of caspase 9 and caspase 3 was also observed. These results indicate that triptolide induced cytotoxicity in L-02 cells by apoptosis, which is mediated through mitochondrial pathway.