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Bilirubin-induced ER stress contributes to the inflammatory response and apoptosis in neuronal cells

  • Organ Toxicity and Mechanisms
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Abstract

Unconjugated bilirubin (UCB) in newborns may lead to bilirubin neurotoxicity. Few studies investigated the activation of endoplasmic reticulum stress (ER stress) by UCB. We performed an in vitro comparative study using undifferentiated SH-SY5Y, differentiated GI-ME-N neuronal cells and human U87 astrocytoma cells. ER stress and its contribution to inflammation and apoptosis induced by UCB were analyzed. Cytotoxicity, ER stress and inflammation were observed only in neuronal cells, despite intracellular UCB accumulation in all three cell types. UCB toxicity was enhanced in undifferentiated SH-SY5Y cells and correlated with a higher mRNA expression of pro-apoptotic CHOP. Mouse embryonic fibroblast knockout for CHOP and CHOP siRNA-silenced SH-SY5Y increased cells viability upon UCB exposure. In SH-SY5Y, ER stress inhibition by 4-phenylbutyric acid reduced UCB-induced apoptosis and decreased the cleaved forms of caspase-3 and PARP proteins. Reporter gene assay and PERK siRNA showed that IL-8 induction by UCB is transcriptionally regulated by NFкB and PERK signaling. These data suggest that ER stress has an important role in the UCB-induced inflammation and apoptosis, and that targeting ER stress may represent a potential therapeutic approach to decrease UCB-induced neurotoxicity.

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Acknowledgments

Authors want to thank Melania Zanchetta for performing plasmid amplification. Authors also thank AREA Science Park for supporting Mohammed Qaisiya fellowship. This work was supported in part by and intramural grant from Fondazione Italiana Fegato, ONLUS and by grant RVO VFN64165 from the Czech Ministry of Health, and by grant KONTAKT LH15097 from the Czech Ministry of Education.

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Authors and Affiliations

  1. Fondazione Italiana Fegato ONLUS, Italian Liver Foundation ONLUS, AREA Science Park Basovizza Bldg Q, 34149, Trieste, Italy

    Mohammed Qaisiya, Cristina Brischetto, Claudio Tiribelli & Cristina Bellarosa

  2. Institute of Medical Biochemistry and Laboratory Medicine, 1st Faculty of Medicine, Charles University in Prague, 12000, Prague, Czech Republic

    Jana Jašprová & Libor Vitek

  3. 4th Department of Internal Medicine, 1st Faculty of Medicine, Charles University in Prague, 12000, Prague, Czech Republic

    Libor Vitek

  4. Department of Medical Sciences, University of Trieste, 34149, Trieste, Italy

    Claudio Tiribelli

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  1. Mohammed Qaisiya
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Correspondence to Mohammed Qaisiya.

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Qaisiya, M., Brischetto, C., Jašprová, J. et al. Bilirubin-induced ER stress contributes to the inflammatory response and apoptosis in neuronal cells. Arch Toxicol 91, 1847–1858 (2017). https://doi.org/10.1007/s00204-016-1835-3

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