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Baicalein-Induced Apoptosis via Endoplasmic Reticulum Stress Through Elevations of Reactive Oxygen Species and Mitochondria Dependent Pathway in Mouse–Rat Hybrid Retina Ganglion Cells (N18)

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Abstract

Studies were designed to investigate the effects of baicalein on mouse–rat hybrid retina ganglion cells (N18) to better understand its effect on apoptosis and apoptosis-related genes in vitro. Cell viability, reactive oxygen species (ROS), cytoplasmic Ca2+, mitochondrial membrane potential (MMP), apoptosis induction, and caspases-3 activity were examined by flow cytometric assay. Apoptosis-associated proteins such as p53, Bax, Bcl-2, cytochrome c, and caspase-3 were examined by Western blot. We demonstrated the increase in the levels of p53, Bax, and cytochrome c and decrease in the level of Bcl-2, which are associated with the induction of apoptotic cell death after 24 h treatment with baicalein in N18 cells. Baicalein induced an increase in the cytoplasmic levels of ROS and Ca2+ in 1 h and reached their peak at 3 h, and thereafter a loss of MMP by flow cytometry. We also demonstrated a release of the cytochrome c from mitochondria into cytosol and an activation of caspase-3, which led to the occurrence of apoptosis in N18 cells treated with baicalein by Western blot. Pretreatment was conducted with BAPTA (intracellular calcium chelator) in baicalein-treated cells, the decline of MMP was recovered, and the increase in the level of cytoplasmic Ca2+ was suppressed, and the proportion of apoptosis was also markedly diminished. In conclusion, our data suggests that oxidative stress and cellular Ca2+ modulates the baicalein-induced cell death via a Ca2+-dependent mitochondrial death pathway in N18 cells.

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Authors and Affiliations

  1. Department of Medical Laboratory Science and Biotechnology, Central Taiwan University of Science and Technology, Taichung, Taiwan, ROC

    Yu-Ching Li

  2. Department of Ophthalmology, China Medical University Hospital, Taichung, Taiwan, ROC

    Hui-Ju Lin

  3. Department of Dermatology, Chung Shan Medical University Hospital, Taichung, Taiwan, ROC

    Jen-Hung Yang

  4. Department of Pharmacology, China Medical University, Taichung, Taiwan, ROC

    Jai-Sing Yang

  5. Department of Biochemistry, China Medical University, Taichung, Taiwan, ROC

    Heng-Chien Ho

  6. School of Chinese Medicine, China Medical University, Taichung, Taiwan, ROC

    Shu-Jen Chang

  7. Department of Internal Medicine, China Medical University Hospital, Taichung, Taiwan, ROC

    Te-Chun Hsai

  8. Department of Clinical Pathology, Cheng Hsin Rehabilitation Medical Center, Taipei, Taiwan, ROC

    Hsu-Feng Lu

  9. Department of Pharmacy, Yuan-Lin Kuo Hospital, Yuanlin Town, Changhua County, Taiwan, ROC

    An-Cheng Huang

  10. Department of Biological Science and Technology, China Medical University, Taichung, Taiwan, ROC

    Jing-Gung Chung

  11. Department of Biotechnology, Asia University, Wufeng, Taichung County, Taiwan, ROC

    Jing-Gung Chung

  12. Department of Microbiology, School of Medicine, China Medical University, No 91, Hsueh-Shih Road, Taichung, 404, Taiwan, ROC

    Jing-Gung Chung

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  1. Yu-Ching Li
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Correspondence to Jing-Gung Chung.

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Li, YC., Lin, HJ., Yang, JH. et al. Baicalein-Induced Apoptosis via Endoplasmic Reticulum Stress Through Elevations of Reactive Oxygen Species and Mitochondria Dependent Pathway in Mouse–Rat Hybrid Retina Ganglion Cells (N18). Neurochem Res 34, 418–429 (2009). https://doi.org/10.1007/s11064-008-9799-1

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