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Lipopolysaccharides Upregulate Hepcidin in Neuron via Microglia and the IL-6/STAT3 Signaling Pathway

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Abstract

Neuroinflammation is closely related to brain iron homeostasis. Our previous study demonstrated that lipopolysaccharides (LPS) can regulate expression of iron-regulatory peptide hepcidin; however, the mechanism is undefined. Here, we demonstrated that intracerebroventricular injection of LPS in rat brain upregulated hepcidin and downregulated ferroportin 1 in the cortex and substantia nigra. LPS increased hepcidin expression in neurons only when they were co-cultured with BV-2 microglia, and the upregulation was suppressed by IL-6 neutralizing antibody in vitro. In addition, IL-6 but not IL-1α, IL-1β, or tumor necrosis factor-alpha increased hepcidin expression and signal transducer and activator of transcription 3 (STAT3) phosphorylation in cortical neurons and MES23.5 dopaminergic neurons. These effects were blocked by the STAT3 inhibitor, stattic. Our results show that neurons are the major source of increased hepcidin expression in response to LPS challenge but microglia play a key mediator role by releasing IL-6 and recruiting the STAT3 pathway. We conclude that LPS upregulates hepcidin expression in neurons via microglia and the IL-6/STAT3 signaling pathway.

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Abbreviations

AD:

Alzheimer’s disease

ELISA:

Enzyme-linked immunosorbent assay

FBS:

Fetal bovine serum

Fpn1:

Ferroportin 1

IL-1α:

Interleukin-1 alpha

IL-1β:

Interleukin-1 beta

IL-6:

Interleukin-6

LPS:

Lipopolysaccharides

MAP2:

Microtubule-associated protein 2

PD:

Parkinson’s disease

STAT3:

Signal transducer and activator of transcription 3

TLRs:

Toll-like receptors

TNF-α:

Tumor necrosis factor-alpha

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Acknowledgments

This work was supported by the Key Project Grant of the National Natural Science Foundation of China (NSFC) (31330035), the Competitive Earmarked Grants of The Hong Kong Research Grants Council (GRF 466713), The Hong Kong Health and Medical Research Fund (grant No. 0112014), National Basic Research Program of China (973 Program, 2011CB510004), the General Grant of NSFC (31271132, 31371092), and Direct Grant of the Chinese University of Hong Kong (4054042).

Conflict of Interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Laboratory of Neuropharmacology, Fudan University School of Pharmacy, Shanghai, 201203, China

    Zhong-Ming Qian, Guang Yang & Qian Qian Luo

  2. School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, NT, Hong Kong SAR, China

    Xuan He, Tuo Liang, Ka-Chun Wu, Yik-Chun Yan, Li-Na Lu, Wing-Ho Yung & Ya Ke

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  1. Zhong-Ming Qian
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Correspondence to Zhong-Ming Qian or Ya Ke.

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Supplementary Figure 1

Effects of LPS on hepcidin expression in microglia and astrocytes in the cortex and substantia nigra in rat brain. Typical micrographs showing that the expression of hepcipin in microglia in these two regions was not changed after LPS treatment, revealed by double immunofluorescence labeling for Iba-1 (A). Similarly, the expression of hepcidin in astrocytes in these two regions was also not elevated, revealed by double immunofluorescence labeling for GFAP (B). Scale bar = 50 μm in main picture and 20 μm in inset (n = 4 in control group; n = 5 in LPS treated group). (TIFF 422 kb)

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Supplementrary Figure 2

Effect of LPS on hepcidin expression in substantia nigra dopamine neurons. The expression of hepcidin in the substantia nigra dopamine neurons was elevated after LPS treatment, revealed by double immunofluorescent labeling for tyrosine hydroxylase (TH) Scale bar = 50 μm in main picture and 20 μm in inset (n = 4 in control group; n = 5 in LPS-treated group). (TIFF 175 kb)

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Qian, ZM., He, X., Liang, T. et al. Lipopolysaccharides Upregulate Hepcidin in Neuron via Microglia and the IL-6/STAT3 Signaling Pathway. Mol Neurobiol 50, 811–820 (2014). https://doi.org/10.1007/s12035-014-8671-3

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