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Inhibition of Urea Transporter (UT)-B Modulates LPS-Induced Inflammatory Responses in BV2 Microglia and N2a Neuroblastoma Cells

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Abstract

Urea is the major nitrogen-containing product of protein metabolism, and the urea cycle is intrinsically linked to nitric oxide (NO) production via the common substrate L-arginine. Urea accumulates in the brain in neurodegenerative states, including Alzheimer’s and Huntington’s disease. Urea transporter B (UT-B, SLC14A1) is the primary transport protein for urea in the CNS, identified most abundantly in astrocytes. Moreover, enhanced expression of the Slc14a1 gene has been reported under neurodegenerative conditions. While the role of UT-B in disease pathology remains unclear, UT-B-deficient mice display behavioural impairment coupled with urea accumulation, NO disruption and neuronal loss. Recognising the role of inflammation in neurodegenerative disease pathology, the current short study evaluates the role of UT-B in regulating inflammatory responses. Using the specific inhibitor UTBinh-14, we investigated the impact of UT-B inhibition on LPS-induced changes in BV2 microglia and N2a neuroblastoma cells. We found that UTBinh-14 significantly attenuated LPS-induced production of TNFα and IL-6 from BV2 cells, accompanied by reduced release of NO. While we observed a similar reduction in supernatant concentration of IL-6 from N2a cells, the LPS-stimulated NO release was further augmented by UTBinh-14. These changes were accompanied by a small, but significant downregulation in UT-B expression in both cell types following incubation with LPS, which was not restored by UTBinh-14. Taken together, the current evidence implicates UT-B in regulation of inflammatory responses in microglia and neuronal-like cells. Moreover, our findings offer support for the further investigation of UT-B as a novel therapeutic target for neuroinflammatory conditions.

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Fig. 1
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Data Availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

Code Availability

Not applicable.

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Funding

A portion of this work was supported by the Jack Pickard Research Fund.

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Authors and Affiliations

  1. UCD School of Biomolecular & Biomedical Science, University College Dublin, Dublin 4, Ireland

    Aimée C. Jones & Derek A. Costello

  2. UCD School of Biology & Environmental Science, University College Dublin, Dublin 4, Ireland

    Farhana Pinki & Gavin S. Stewart

  3. UCD Conway Institute, University College Dublin, Dublin 4, Ireland

    Aimée C. Jones & Derek A. Costello

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  2. Farhana Pinki
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  4. Derek A. Costello
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Contributions

A.C.J conducted the experiments, wrote and edited the manuscript. F.P. carried out PCR analysis. G.S.S contributed to study conception and design, edited and revised the manuscript. D.A.C conceived the study, supervised the work, analysed data, wrote and edited the manuscript.

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Correspondence to Derek A. Costello.

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Jones, A.C., Pinki, F., Stewart, G.S. et al. Inhibition of Urea Transporter (UT)-B Modulates LPS-Induced Inflammatory Responses in BV2 Microglia and N2a Neuroblastoma Cells. Neurochem Res 46, 1322–1329 (2021). https://doi.org/10.1007/s11064-021-03283-4

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