Caspase 3 is involved in the apoptosis induced by triptolide in HK-2 cells.
Triptolide, a main active component extracted from the traditional Chinese herbal medicine, Tripterygium wilfordii Hook. f (TWHf), has been shown to possess potent immunosuppressive and anti-inflammatory properties. However, the toxicity of triptolide limits its clinical applications. Here we treated the human proximal tubular epithelial cell line HK-2 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 viability inhibition and annexin V/propidium iodide (PI) staining for apoptosis/necrosis. The activation of caspase 3 was analyzed by Western Blotting. MTT assay showed triptolide inhibited the viability of HK-2 cells in a time- and dose-dependent manner. Flow cytometry assay showed triptolide caused apoptosis rather than necrosis in HK-2 cells by staining with annexin V/PI. Furthermore, the increase of cleaved p17 fragment, an active form of caspase 3, was detected. These results suggested that triptolide is able to cause cytotoxicity on HK-2 cells, and the mechanism of which is associated with caspase 3.
Shu
Bin
B
Jiangsu Center for Drug Screening, China Pharmaceutical University, 24 Tong Jia Xiang, Nanjing 210009, PR China.
Duan
Weigang
W
Yao
Jincheng
J
Huang
Jingfeng
J
Jiang
Zhenzhou
Z
Zhang
Luyong
L
D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000633:toxicity*; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000502:physiology*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004224:Diterpenes / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D004852:Epoxy Compounds / Q000633:toxicity; D006801:Humans; D007687:Kidney Tubules, Proximal / Q000187:drug effects* / Q000473:pathology; D010616:Phenanthrenes / Q000633:toxicity*
D000894:Anti-Inflammatory Agents, Non-Steroidal; D004224:Diterpenes; D004852:Epoxy Compounds; D010616:Phenanthrenes; C001899:triptolide; D053148:Caspase 3