Saikosaponin d induces cell death through caspase-3-dependent, caspase-3-independent and mitochondrial pathways in mammalian hepatic stellate cells

BMC Cancer

Research Article

Wen-Chuan Hsieh¹  Ming-Feng Chen²  Li-Yen Shiu³  Pei-Shan Liu⁴  S. Joseph Huang⁵  Chao-Cheng Huang⁶  Kun-I Lin⁷  Chang-Han Chen⁸  Ching-Wen Liu⁹ 

[1] Department of Biological Science & Technology, I-SHOU University, Kaohsiung, Taiwan;Department of Gastroenterology and Hepatology, E-DA Hospital, Kaohsiung, Taiwan;Graduate Institute of Integrated Medicine, China Medical University, Taichung, Taiwan;Department of Medical Research, E-Da Hospital, Kaohsiung, Taiwan;Cell Therapy and Research Center, Department of Medical Research, E-Da Cancer Hospital, Kaohsiung, Taiwan;Department of Microbiology, Soochow University, Shihlin, Taipei, Taiwan;Department of Obstetrics and Gynecology, E-Da Hospital, I-Shou University, Kaohsiung, Taiwan;Department of Obstetrics and Gynecology, University of South Florida, College of Medicine, Tampa, FL, USA;Department of Pathology, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung, Taiwan;Tissue Bank and Biobank, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan;Departments of Obstetrics & Gynecology, Chang Bing Show Chwan Memorial Hospital, Lukang Zhen, Changhua County, Taiwan;Institute for Translational Research in Biomedicine, Kaohsiung Chang Gung Memorial Hospital, 123 Ta-Pei Road, Niaosong District, Kaohsiung City, Taiwan;Department of Applied Chemistry, National Chi Nan University, Nantou, Taiwan;Center for Infectious Disease and Cancer Research, Kaohsiung Medical University, Kaohsiung, Taiwan;School of Pharmacy, Kaohsiung Medical University, Kaohsiung, Taiwan;

关键词: Mitochondrial Membrane Potential;    Cytosolic Fraction;    Oxygen Consumption Rate;    Extracellular Acidification;    Apoptotic Factor;   

DOI  :  10.1186/s12885-016-2599-0

 received in 2016-03-28, accepted in 2016-07-22,  发布年份 2016

来源: Springer

【 摘 要 】

BackgroundSaikosaponin d (SSd) is one of the main active triterpene saponins in Bupleurum falcatum. It has a steroid-like structure, and is reported to have pharmacological activities, including liver protection in rat, cell cycle arrest and apoptosis induction in several cancer cell lines. However, the biological functions and molecular mechanisms of mammalian cells under SSd treatment are still unclear.MethodsThe cytotoxicity and apoptosis of hepatic stellate cells (HSCs) upon SSd treatment were discovered by MTT assay, colony formation assay and flow cytometry. The collage I/III, caspase activity and apoptotic related genes were examined by quantitative PCR, Western blotting, immunofluorescence and ELISA. The mitochondrial functions were monitored by flow cytometry, MitoTracker staining, ATP production and XF24 bioenergetic assay.ResultsThis study found that SSd triggers cell death via an apoptosis path. An example of this path might be typical apoptotic morphology, increased sub-G1 phase cell population, inhibition of cell proliferation and activation of caspase-3 and caspase-9. However, the apoptotic effects induced by SSd are partially blocked by the caspase-3 inhibitor, Z-DEVD-FMK, suggesting that SSd may trigger both HSC-T6 and LX-2 cell apoptosis through caspase-3-dependent and independent pathways. We also found that SSd can trigger BAX and BAK translocation from the cytosol to the mitochondria, resulting in mitochondrial function inhibition, membrane potential disruption. Finally, SSd also increases the release of apoptotic factors.ConclusionsThe overall analytical data indicate that SSd-elicited cell death may occur through caspase-3-dependent, caspase-3-independent and mitochondrial pathways in mammalian HSCs, and thus can delay the formation of liver fibrosis by reducing the level of HSCs.

【 授权许可】

CC BY   
© The Author(s). 2016