Caspase Inhibition Augmented Oridonin-Induced Cell Death in Murine Fibrosarcoma L 929 by Enhancing Reactive Oxygen Species Generation
Oridonin, a diterpenoid isolated from Rabdosia rubescences, has been reported to have antitumor effects. In this study, the growth-inhibitory activity of oridonin for L929 cells was exerted in a time-and dose-dependent manner. After treatment with oridonin for 24 h, L929 cells underwent both apoptos...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2008-01-01
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| Series: | Journal of Pharmacological Sciences |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1347861319313714 |
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| author | Jin-Nan Wu Jian Huang Jia Yang Shin-Ichi Tashiro Satoshi Onodera Takashi Ikejima |
| author_facet | Jin-Nan Wu Jian Huang Jia Yang Shin-Ichi Tashiro Satoshi Onodera Takashi Ikejima |
| author_sort | Jin-Nan Wu |
| collection | DOAJ |
| description | Oridonin, a diterpenoid isolated from Rabdosia rubescences, has been reported to have antitumor effects. In this study, the growth-inhibitory activity of oridonin for L929 cells was exerted in a time-and dose-dependent manner. After treatment with oridonin for 24 h, L929 cells underwent both apoptosis and necrosis as measured by an lactate dehydrogenase (LDH) activity-based assay. A rapid generation of reactive oxygen species (ROS) was triggered by oridonin, and subsequently up-regulation of phospho-p53 (ser 15) expression and an increased expression ratio of Bax/Bcl-2 was observed. Furthermore, there was a significant fall in mitochondrial membrane potential (MMP) and increase in caspase-3 activity after exposure to oridonin for 24 h. Surprisingly, the pan-caspase inhibitor z-VAD-fmk and caspase 3 inhibitor z-DEVD-fmk rendered L929 cells more sensitive to oridonin, rather than preventing oridonin-induced cell death. Oridonin and z-VAD-fmk co-treatment not only resulted in an even higher ROS production, but also made a more significant reduction in the MMP. Pretreatment of ROS scavenger N-acetylcysteine (NAC) led to a complete inhibition of oridonin-induced cell death, intracellular ROS generation, and MMP collapse. NAC treatment also reversed the potentiation of cell death by the pan-caspase inhibitor z-VAD-fmk. Taken together, these observations showed that oridonin-induced cell death in L929 cells involved intracellular ROS generation, activation of phospho-p53 (ser 15), and up-regulation of the Bax/Bcl-2 ratio; and the augmented cell death by z-VAD-fmk was dependent on an increased ROS production. Keywords:: oridonin, reactive oxygen species (ROS), Bcl-2, L929, apoptosis |
| first_indexed | 2024-12-12T00:40:43Z |
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| institution | Directory Open Access Journal |
| issn | 1347-8613 |
| language | English |
| last_indexed | 2024-12-12T00:40:43Z |
| publishDate | 2008-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Pharmacological Sciences |
| spelling | doaj.art-1664b30923e540aa873a035e2bff86722022-12-22T00:44:15ZengElsevierJournal of Pharmacological Sciences1347-86132008-01-0110813239Caspase Inhibition Augmented Oridonin-Induced Cell Death in Murine Fibrosarcoma L 929 by Enhancing Reactive Oxygen Species GenerationJin-Nan Wu0Jian Huang1Jia Yang2Shin-Ichi Tashiro3Satoshi Onodera4Takashi Ikejima5China-Japan Research Institute of Medical Pharmaceutical Sciences, Shenyang 110016, ChinaDepartment of Phytochemistry, Shenyang Pharmaceutical University, Shenyang 110016, ChinaDepartment of Phytochemistry, Shenyang Pharmaceutical University, Shenyang 110016, ChinaDepartment of Clinical and Biomedical Sciences, Showa Pharmaceutical University, Tokyo 194-8543, JapanDepartment of Clinical and Biomedical Sciences, Showa Pharmaceutical University, Tokyo 194-8543, JapanChina-Japan Research Institute of Medical Pharmaceutical Sciences, Shenyang 110016, China; Corresponding author. ikejimat@vip.sina.comOridonin, a diterpenoid isolated from Rabdosia rubescences, has been reported to have antitumor effects. In this study, the growth-inhibitory activity of oridonin for L929 cells was exerted in a time-and dose-dependent manner. After treatment with oridonin for 24 h, L929 cells underwent both apoptosis and necrosis as measured by an lactate dehydrogenase (LDH) activity-based assay. A rapid generation of reactive oxygen species (ROS) was triggered by oridonin, and subsequently up-regulation of phospho-p53 (ser 15) expression and an increased expression ratio of Bax/Bcl-2 was observed. Furthermore, there was a significant fall in mitochondrial membrane potential (MMP) and increase in caspase-3 activity after exposure to oridonin for 24 h. Surprisingly, the pan-caspase inhibitor z-VAD-fmk and caspase 3 inhibitor z-DEVD-fmk rendered L929 cells more sensitive to oridonin, rather than preventing oridonin-induced cell death. Oridonin and z-VAD-fmk co-treatment not only resulted in an even higher ROS production, but also made a more significant reduction in the MMP. Pretreatment of ROS scavenger N-acetylcysteine (NAC) led to a complete inhibition of oridonin-induced cell death, intracellular ROS generation, and MMP collapse. NAC treatment also reversed the potentiation of cell death by the pan-caspase inhibitor z-VAD-fmk. Taken together, these observations showed that oridonin-induced cell death in L929 cells involved intracellular ROS generation, activation of phospho-p53 (ser 15), and up-regulation of the Bax/Bcl-2 ratio; and the augmented cell death by z-VAD-fmk was dependent on an increased ROS production. Keywords:: oridonin, reactive oxygen species (ROS), Bcl-2, L929, apoptosishttp://www.sciencedirect.com/science/article/pii/S1347861319313714 |
| spellingShingle | Jin-Nan Wu Jian Huang Jia Yang Shin-Ichi Tashiro Satoshi Onodera Takashi Ikejima Caspase Inhibition Augmented Oridonin-Induced Cell Death in Murine Fibrosarcoma L 929 by Enhancing Reactive Oxygen Species Generation Journal of Pharmacological Sciences |
| title | Caspase Inhibition Augmented Oridonin-Induced Cell Death in Murine Fibrosarcoma L 929 by Enhancing Reactive Oxygen Species Generation |
| title_full | Caspase Inhibition Augmented Oridonin-Induced Cell Death in Murine Fibrosarcoma L 929 by Enhancing Reactive Oxygen Species Generation |
| title_fullStr | Caspase Inhibition Augmented Oridonin-Induced Cell Death in Murine Fibrosarcoma L 929 by Enhancing Reactive Oxygen Species Generation |
| title_full_unstemmed | Caspase Inhibition Augmented Oridonin-Induced Cell Death in Murine Fibrosarcoma L 929 by Enhancing Reactive Oxygen Species Generation |
| title_short | Caspase Inhibition Augmented Oridonin-Induced Cell Death in Murine Fibrosarcoma L 929 by Enhancing Reactive Oxygen Species Generation |
| title_sort | caspase inhibition augmented oridonin induced cell death in murine fibrosarcoma l 929 by enhancing reactive oxygen species generation |
| url | http://www.sciencedirect.com/science/article/pii/S1347861319313714 |
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