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Downregulation of microsomal glutathione-S-transferase 1 modulates protective mechanisms in differentiated PC12 cells

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Abstract

Microsomal glutathione-S-transferase 1 (Mgst1) plays a specific role in protection of cells against oxidative stress. In this study, we assayed the effect of Mgst1 downregulation on cells behavior using differentiated PC12 line, a widely accepted neuronal model system. We have developed stable transfected cells with downregulated Mgst1 (PC12_M), which were differentiated with 1 mM dibutyryl-cAMP (db-cAMP). Mgst1 reduction induced necrosis, decreased ATP amount, and increased thiobarbituric acid reacting substances (TBARS) content. However, in PC12_M cell population, we detected more intensive neuritogenesis than that in mock-transfected cells. Interestingly, total glutathione as well as GSH level were significantly higher than those in control PC12 line. Real-time PCR and Western blot analyses showed elevated expression of enzymes involved in glutathione metabolism—a rate-limiting γ-glutamylcysteine ligase and glutathione reductase. The present study shows for the first time that under stress conditions induced by Mgst1 downregulation, a rescue pathway can be activated and thereby enables differentiated PC12 cells to survive. Since Mgst1expression was reported to decline with age, our results could represent a putative adaptive process during aging. It could also be an early mechanism protecting neuronal cells against some neurodegenerative insults.

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Acknowledgments

We thank Bozena Ferenc and Malwina Lisek for technical assistance. This work was supported by the grants 502-03/6-086-02/502-64-003 and 503/6-086-02/503-01 from the Medical University of Lodz.

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Authors and Affiliations

  1. Department of Molecular Neurochemistry, Medical University, 6/8 Mazowiecka Str., 92-215, Lodz, Poland

    Monika Sobczak, Tomasz Boczek, Antoni Kowalski & Ludmila Zylinska

  2. Department of Molecular and Medical Biophysics, Medical University, 6/8 Mazowiecka Str., 92-215, Lodz, Poland

    Magdalena Wiktorska & Jolanta Niewiarowska

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  1. Monika Sobczak
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  2. Tomasz Boczek
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  3. Antoni Kowalski
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  4. Magdalena Wiktorska
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  5. Jolanta Niewiarowska
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  6. Ludmila Zylinska
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Correspondence to Ludmila Zylinska.

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Sobczak, M., Boczek, T., Kowalski, A. et al. Downregulation of microsomal glutathione-S-transferase 1 modulates protective mechanisms in differentiated PC12 cells. J Physiol Biochem 70, 375–383 (2014). https://doi.org/10.1007/s13105-014-0312-9

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  • DOI: https://doi.org/10.1007/s13105-014-0312-9

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