Tetramethylpyrazine Protects Blood-Spinal Cord Barrier Integrity by Modulating Microglia Polarization Through Activation of STAT3/SOCS3 and Inhibition of NF-кB Signaling Pathways in Experimental Autoimmune Encephalomyelitis Mice (original) (raw)

Abstract

We previously reported that tetramethylpyrazine (TMP) alleviates experimental autoimmune encephalomyelitis (EAE) by decreasing glia activation. Activated microglia has been shown to mediate blood-spinal cord barrier (BSCB) disruption, which is a primary and continuous pathological characteristic of multiple sclerosis (MS). Therefore, in this study, we further investigated whether TMP protects the BSCB integrity by inhibition of glia activation to alleviate EAE. Extravasation of evans blue was used to detect the BSCB disruption. Tumor necrosis factor-α (TNF-α)/interlukine-1β (IL-1β) and interlukine-4 (IL-4)/interlukine-10 (IL-10) were determined by enzyme-linked immunosorbent assay. BV2 glial cells stimulated by interferon-γ (IFN-γ) were co-cultured with human brain microvascular endothelial cells to investigate the effect of TMP on the BSCB disruption. Flow cytometry was used to analyze the microglia phenotype. Western blot was performed to reveal the signaling pathways involved in the microglia activation. In this study, most importantly, we found that TMP protects the BSCB integrity by modulating microglia polarization from M1 phenotype to M2 phenotype through activation of STAT3/SOCS3 and inhibition of NF-к_B signaling pathways. Moreover, TMP significantly preserves the tight junction proteins, reduces the secretion of pro-inflammatory cytokines (TNF-α, IL-1_β) and increases the secretion of anti-inflammatory cytokines (IL-4, IL-10) from IFN-γ-stimulated BV2 microglia cells. Consequently, protection of the BSCB integrity leads to alleviation of clinical symptoms and demyelination in EAE mice. Therefore, TMP might be an effective therapeutic agent for cerebral disorders with BBB or BSCB disruption, such as ischemic stroke, MS, and traumatic brain injury.

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Acknowledgements

This study was supported by The National Natural Science Foundation of China [81601049, 21402010], Nature Science Foundation of Shandong Province [ZR2019MB032], Key Research and Development Program of Shandong Province [2018GSF118129].

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Author notes

1. Lianshuang Zhang, Xueyan Lu, and Lihua Gong have contributed equally to this work.

Authors and Affiliations

1. Department of Histology and Embryology, Binzhou Medical University, Yantai, 264003, People’s Republic of China
   Lianshuang Zhang, Xueyan Lu, Lihua Gong, Linlu Cui, Hongqin Zhang, Wei Zhao, Pengyu Jiang & Yun Hou
2. The Key Laboratory of Prescription Effect and Clinical Evaluation of State Administration of Traditional Chinese Medicine of China, Binzhou Medical University, Yantai, 264003, People’s Republic of China
   GuiGe Hou

Authors

1. Lianshuang Zhang
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2. Xueyan Lu
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3. Lihua Gong
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4. Linlu Cui
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5. Hongqin Zhang
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6. Wei Zhao
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7. Pengyu Jiang
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8. GuiGe Hou
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9. Yun Hou
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Contributions

LSZ, XYL and LHG performed the experiments and collected data. LLC and HQZ carried out the clinical scores of the EAE mice. WZ and PYJ did the statistic analysis, GGH and YH designed the experiment and edited the manuscript. All authors approved the final manuscript.

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Correspondence toGuiGe Hou or Yun Hou.

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Zhang, L., Lu, X., Gong, L. et al. Tetramethylpyrazine Protects Blood-Spinal Cord Barrier Integrity by Modulating Microglia Polarization Through Activation of STAT3/SOCS3 and Inhibition of NF-_к_B Signaling Pathways in Experimental Autoimmune Encephalomyelitis Mice.Cell Mol Neurobiol 41, 717–731 (2021). https://doi.org/10.1007/s10571-020-00878-3

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• Received: 05 February 2020
• Accepted: 12 May 2020
• Published: 18 May 2020
• Issue Date: May 2021
• DOI: https://doi.org/10.1007/s10571-020-00878-3

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