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Bilirubin Induces A1-Like Reactivity of Astrocyte

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Abstract

Astrocytes play an important role in the pathogenesis of bilirubin neurotoxicity, and activated astrocytes might be potential mediators of neuroinflammation processes contributing to neuronal cell death and tissue injury. Recent studies have reported that activated microglia induce two types of reactive astrocytes. A1 astrocytes could cause neuronal death and synaptic damage, as well as impaired phagocytosis. Therefore, the purpose of this study was to investigate whether unconjugated bilirubin (UCB)-induced A1-like astrocytes take on a neuroinflammation type and the underlying regulatory mechanisms. In this study, primary cortical astrocytes were treated with UCB in vitro. We detected the expression of complement component 3 (C3), S100 calcium binding protein A10 (S100A10), nuclear factor kappa B (NF-κB), NLR family pyrin domain containing 3 (NLRP3), activated caspase-1, gasdermin D N-terminal (GSDMD-N), PSD95, synaptophysin (SYP), the transcription levels of interleukin (IL)-1β and IL-18, and the survival rate of astrocytes after UCB treatment. The results showed that an increase in C3 was accompanied by a decrease in S100A10, and that A1-like astrocytes were functionally expressed after UCB stimulation. Meanwhile, the NF-κB and caspase-1 pathways were activated after UCB stimulation. After adding the NF-κB-specific inhibitor trans-activator of transcriptional-NEMO-binding domain (TAT-NBD) and caspase-1 specific inhibitor VX-765, the survival rate of astrocytes and neurons increased, whereas the protein expression of C3, NF-κB, NLRP3, activated caspase-1, and GSDMD-N decreased, and the mRNA levels of IL-1β and IL-18 reduced. Thus, we concluded that UCB stimulates the activation of A1-like astrocytes. Inhibition of NF-κB and caspase-1 alleviated A1-like astrocytes and exerted anti-inflammatory protective effects.

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Data Availability

All data generated or analysed during this study are available from the corresponding author upon reasonable request.

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Funding

This work was supported by the National Natural Science Foundation of China (Grant 81971426), the Project of Basic and Frontier Research Plan of Chongqing (Grant CSTC2018jcyjAX0284) and the Postgraduate Research Innovation Project of Chongqing (Grant CYS21227).

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Authors and Affiliations

  1. Department of Neonatology, Children’s Hospital of Chongqing Medical University, National Clinical Research Center for Child Health and Disorders, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Chongqing Key Laboratory of Child Infection and Immunity, Chongqing, 400014, China

    Siyu Li, Hongmei Huang, Yan Zhang, Ling Li & Ziyu Hua

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  1. Siyu Li
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  2. Hongmei Huang
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  3. Yan Zhang
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  4. Ling Li
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Contributions

ZH and SL conceived and designed the study. SL, HH, YZ and LL performed the experiments. SL and ZH participated in analysis and interpretation of data. SL and ZH wrote the manuscript. All authors read and approved the final manuscript.

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Correspondence to Ziyu Hua.

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The experimental process was strictly in accordance with the Ethics Committee of Chongqing Medical University and the Chinese Measures for the Use and Management of Experimental Animals.

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Li, S., Huang, H., Zhang, Y. et al. Bilirubin Induces A1-Like Reactivity of Astrocyte. Neurochem Res 48, 804–815 (2023). https://doi.org/10.1007/s11064-022-03810-x

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  • DOI: https://doi.org/10.1007/s11064-022-03810-x

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