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-кB signaling pathways. Moreover, TMP significantly preserves the tight junction proteins, reduces the secretion of pro-inflammatory cytokines (TNF-α, IL-1β) 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.
Similar content being viewed by others
References
Almutari MM, Gong C, Xu YG, Chang Y, Shi H (2016) Factors controlling permeability of the blood-brain barrier. Cell Mol Life Sci 73:57–77. https://doi.org/10.1007/s00018-015-2050-8
Al Zoubi S, Chen J, Murphy C, Martin L, ChiazzaThiemermann FC (2018) Linagliptin attenuates the cardiac dysfunction associated with experimental sepsis in mice with pre-existing type2 diabetes by inhibiting NF-κB. Front Immunol 9:2996. https://doi.org/10.3389/fimmu.2018.02996
Bai XY, Wang XF, Zhang LS, Du PC, Cao Z, Hou Y (2018) Tetramethylpyrazine ameliorates experimental autoimmune encephalomyelitis by modulating the inflammatory response. Biochem Biophys Res Commun 503:1968–1972. https://doi.org/10.1016/j.bbrc.2018.07.143
Bartanusz V, Jezova D, Alajajian B, Digicaylioglu M (2017) The blood-spinal cord barrier: morphology and clinical implications. Ann Neurol 70:194–206. https://doi.org/10.1002/ana.22421
Bennett J, Basivireddy J, Kollar A, Biron KE, Reickmann P, Jefferies WA, Mc Quaid S (2010) Blood-brain barrier disruption and enhanced vascular permeability in the multiple sclerosis model EAE. J Neuroimmunol 229:180–191. https://doi.org/10.1016/j.jneuroim.2010.08.011
Cevey ÁC, Penas FN, Alba Soto D, Mirkin GA, Goren NB (2019) IL-10/STAT3/SOCS3 axis is involved in the anti-inflammatory effect of benznidazole. Front Immunol 10:1267. https://doi.org/10.3389/fimmu.2019.01267
Chang Y, Hsiao G, Chen SH, Chen YC, Lin JH, Chou DS, Sheu JR (2007) Tetramethylpyrazine suppresses HIF-1 alpha, TNF-alpha, and activated caspase-3 expression in middle cerebral artery occlusion-induced brain ischemia in rats. Acta Pharmacol Sin 28:327–333. https://doi.org/10.1111/j.1745-7254.2007.00514.x
Clarner T, Diederichs F, Berger K, Denecke B, Gan L, Van der Valk P, Beyer C, Amor S, Kipp M (2012) Myelin debris regulates inflammatory responses inan experimental demyelination animal model and multiple sclerosislesions. Glia 60:1468–1480. https://doi.org/10.1002/glia.22367
Fan K, Ma J (2019) Cathepsin C promotes microglia M1 polarization and aggravates neuroinflammation via activation of Ca2+-dependent PKC/p38MAPK/NF-κB pathway. J Neuroinflamm 16:10. https://doi.org/10.1186/s12974-019-1398-3
Fan L, Wang K, Shi Z, Die J, Wang C, Dang X (2011) Tetramethylpyrazine protects spinal cord and reduces inflammation in a rat model of spinal cord ischemia/reperfusion injury. J Vasc Surg 54:192–200. https://doi.org/10.1016/j.jvs.2010.12.030
Feng L, Ke N, Cheng F, Guo Y, Li S, Li Q, Li Y (2011) The protective mechanism of ligustrazine against renal ischemia/reperfusion injury. J Surg Res 166:298–305. https://doi.org/10.1016/j.jss.2009.04.005
Galbraith NJ, Manek S, Walker S, Bishop C, Carter JV, Cahill M, Gardner SA, Polk HC Jr, Galandiuk S (2018) The effect of IκK-16 on lipopolysaccharide-induced impaired monocytes. Immunobiology 223:365–373. https://doi.org/10.1016/j.imbio.2017.10.045
Gong P, Zhang Z, Zou Y, Tian Q, Han S, Xu Z, Liao J, Gao L, Chen Q, Li M (2019) Tetramethylpyrazine attenuates blood-brain barrier disruption in ischemia/reperfusion injury through the JAK/STAT signaling pathway. Eur J Pharmacol 854:289–297. https://doi.org/10.1016/j.ejphar.2019.04.028
Graeber MB, Streit WJ (2010) Microglia: biology and pathology. Acta Neuropathol 119:89–105. https://doi.org/10.1007/s00401-009-0622-0
Gu X, Wei ZZ, Espinera A, Lee JH, Ji X, Wei L, Dix TA, Yu SP (2015) Pharmacologically induced hypothermia attenuates traumatic brain injury in neonatal rats. Exp Neurol 267:135–142. https://doi.org/10.1016/j.expneurol.2015.02.029
Hou Y, Heon RC, Jun JA, Kim SM, Jeong CH, Jeun SS (2014) Interferon β-secreting mesenchymal stem cells combined with minocycline attenuate experimental autoimmune encephalomyelitis. J Neuroimmunol 274:20–27. https://doi.org/10.1016/j.jneuroim.2014.06.001
Hou Y, Ryu CH, Park KY, Kim SM, Jeong CH, Jeun SS (2013) Effective combination of human bone marrow mesenchymal stem cells and minocycline in experimental autoimmune encephalomyelitis mice. Stem Cell Res Ther 4:77. https://doi.org/10.1186/scrt228
Kao TK, Ou YC, Kuo JS, Chen WY, Liao SL, Wu CW, Chen CJ, Ling NN, Zhang YH, Peng WH (2006) Neuroprotection by tetramethylpyrazine against ischemic brain injury in rats. Neuroche Int 48:166–176. https://doi.org/10.1016/j.neuint.2005.10.008
Kreutzberg GW (1996) Microglia: a sensor for pathological events in the CNS. Trends Neurosci 19:312–318. https://doi.org/10.1016/0166-2236(96)10049-7
Li H, Wang W, Wang G, Hou Y, Xu F, Liu R, Wang F, Xue J, Hu T, Luan X (2015) Interferon-γand tumor necrosis factor-α promote the ability of humanplacenta-derived mesenchymal stromal cells to express programmed deathligand-2 and induce the differentiation of CD4(+)interleukin-10(+)and CD8(+)interleukin-10(+)Treg subsets. Cytotherapy 17:1560–1571. https://doi.org/10.1016/j.jcyt.2015.07.018
Liu Q, Zhang Y, Liu S, Liu Y, Yang X, Liu G, Shimizu T, Ikenaka K, Laffer B, Bauer D et al (2019) Loss of IL-10 promotes differentiation of microglia to a M1 phenotype. Front Cell Neurosci 13:430. https://doi.org/10.3389/fncel.2019.00430
Lv L, Jiang SS, Xu J, Gong JB, Cheng Y (2012) Protective effect of ligustrazine against myocardial ischaemia/reperfusion in rats: the role of endothelial nitric oxide synthase. Clin Exp Pharmacol Physiol 39:20–27. https://doi.org/10.1111/j.1440-1681.2011.05628.x
Meireles M, Marques C, Norberto S, Santos P, Fernandes I, Mateus N, Faria A, Calhau C (2016) Anthocyanin effects on microglia M1/M2 phenotype: consequence on neuronal fractalkine expression. Behav Brain Res 305:223–228. https://doi.org/10.1016/j.bbr.2016.03.010
Nakajima K, Kohsaka S (1993) Functional roles of microglia in the brain. Neurosci Res 17:187–203. https://doi.org/10.1016/0168-0102(93)90047-t
Nishioku T, Matsumoto J, Dohgu S, Sumi N, Miyao K, Takata F, Shuto H, Yamauchi A, Kataoka Y (2010) Tumor necrosis factor-alpha mediates the blood-brain barrier dysfunction induced by activated microglia in mouse brain microvascular endothelial cells. J Pharmacol Sci 112:251–254. https://doi.org/10.1254/jphs.09292sc
Nuttall RK, Silva C, Hader W, Bar-Or A, Patel KD, Edwards DR, Yong VW (2007) Metalloproteinases are enriched in microglia compared with leukocytes and they regulate cytokine levels in activated microglia. Glia 55:516–526. https://doi.org/10.1002/glia.20478
Petty MA, Lo EH (2003) Junctional complexes of the blood-brain barrier: permeability changes in neuroinflammation. Prog Neurobiol 68:311–323. https://doi.org/10.1016/s0301-0082(02)00128-4
Salter MW, Beggs S (2014) Sublime microglia: expanding roles for the guardians of the CNS. Cell 158:15–24. https://doi.org/10.1016/j.cell.2014.06.008
Streit WJ, Xue QS, Tischer J, Bechmann I (2014) Microglial pathology. Acta Neuropathol Commun 2:142. https://doi.org/10.1186/s40478-014-0142-6
Wang A, Zhu G, Qian P (2017) Zhu T (2017) Tetramethylpyrazine reduces blood-brain barrier permeability associated with enhancement of peripheral cholinergic anti-inflammatory effects for treatingtraumatic brain injury. Exp Ther Med 14(3):2392–2400. https://doi.org/10.3892/etm.2017.4754
Xiao X, Liu Y, Qi C, Qiu F, Chen X, Zhang J, Yang P (2010) Neuroprotection and enhanced neurogenesis by tetramethylpyrazine in adult rat brain after focal ischemia. Neurol Res 32:47–555. https://doi.org/10.1179/174313209X414533
Yan G, Zhu Z, Jin L, Chen J, Xie H, Miozzi J, Lei F, Wei X, Pan J (2018) Study on the quality evaluation of compound danshen preparations based on the xCELLigence real-time cell-based assay and pharmacodynamic authentication. Molecules 23:2090. https://doi.org/10.3390/molecules23092090
Yang G, Qian C, Wang N, Lin C, Wang Y, Wang G, Piao X (2017) Tetramethylpyrazine protects against oxygen-glucose deprivation-induced brain microvascular endothelial cells injury via Rho/Rho-kinase signaling pathway. Cell Mol Neurobiol 37(4):619–633. https://doi.org/10.1007/s10571-016-0398-4
Yang Y, Salayandia VM, Thompson JF, Yang LY, Estrada EY, Yang Y (2015) Attenuation of acute stroke injury in rat brain by minocycline promotes blood-brain barrier remodeling and alternative microglia/macrophage activation during recovery. J Neuroinflamm 12:26. https://doi.org/10.1186/s12974-015-0245-4
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].
Author information
Authors and Affiliations
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.
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interests.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
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-кB Signaling Pathways in Experimental Autoimmune Encephalomyelitis Mice. Cell Mol Neurobiol 41, 717–731 (2021). https://doi.org/10.1007/s10571-020-00878-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10571-020-00878-3