AGK2 Alleviates Lipopolysaccharide Induced Neuroinflammation through Regulation of Mitogen-Activated Protein Kinase Phosphatase-1

  • Fangzhou Jiao
  • Yao Wang
  • Wenbin Zhang
  • Haiyue Zhang
  • Qian Chen
  • Luwen Wang
  • Chunxia Shi
  • Zuojiong GongEmail author


Neuroinflammation is associated with the progression of multiple neurological diseases. Many studies show that SIRT2 involves in multiple inflammatory processes. While, the mechanisms remain unclear. The purpose of this study was to explore the effect of SIRT2 inhibitor AGK2 on inflammatory responses and MAPK signaling pathways in LPS activated microglia in vitro and in vivo. The effect of AGK2 on cell viability of BV2 microglial cells was detected by CCK-8 assay. The expression of inflammatory cytokine iNOS was analyzed by western blotting and immunofluorescence. The mRNA expressions of iNOS, TNF-α, and IL-1β were detected by real-time polymerase chain reaction (RT-PCR). The SIRT2, phospho-P38, P38, phospho-JNK, JNK, phospho-ERK, ERK, α-tubulin, and acetyl-α-tubulin were analyzed by western blotting respectively. The interaction between SIRT2 and MKP-1 was measured by Co-immunoprecipitation (Co-IP) assay. Double immunofluorescent staining was performed to detect the expressions of CD11b and iNOS or SIRT2 in brain tissues. We found that AGK2 could suppress LPS-induced inflammatory cytokines (iNOS, TNF-α, and IL-1β) expression levels in BV2 microglial cells. Moreover, it could effectively reduce the expression of SIRT2 and increase the acetylation of α-tubulin in LPS activated BV2 microglial cells and LPS induced mice neuroinflammation. In addition, our results showed that AGK2 could reduce the increase of phosphorylation p38, JNK, and ERK after LPS challenge. Co-IP results showed that there was no direct interaction between MKP-1 and SIRT2. However, AGK2 by inhibition of SIRT2 could increase the expression of MKP-1. Furthermore, AGK2 could inhibit the activation of BV2 microglia and expression of iNOS and SIRT2 in LPS treated mice brain tissue. Taken together, our results suggested that AGK2 might alleviate lipopolysaccharide induced neuroinflammation through regulation of mitogen-activated protein kinase phosphatase-1.

Graphical abstract


AGK2 SIRT2 Neuroinflammation Microglia MKP-1 



This study was supported by a grant from the National Natural Science Foundation of China (No. 81870413).

Author’s Contributions

Fangzhou Jiao and Zuojiong Gong designed research; Fangzhou Jiao performed research; Yao Wang, Wenbin Zhang, Haiyue Zhang, Qian Chen and Luwen Wang analyzed and interpreted the data. Chunxia Shi performed supplementary experiment and corrected English errors in paper. Fangzhou Jiao and Zuojiong Gong wrote the paper.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.


  1. Brown GC (2007) Mechanisms of inflammatory neurodegeneration: iNOS and NADPH oxidase. Biochem Soc Trans 35(Pt 5):1119–1121CrossRefGoogle Scholar
  2. Cao W, Bao C, Padalko E, Lowenstein CJ (2008) Acetylation of mitogen-activated protein kinase phosphatase-1 inhibits toll-like receptor signaling. J Exp Med 205(6):1491–1503. CrossRefPubMedPubMedCentralGoogle Scholar
  3. Catorce MN, Gevorkian G (2016) LPS-induced murine Neuroinflammation model: main features and suitability for pre-clinical assessment of Nutraceuticals. Curr Neuropharmacol 14(2):155–164CrossRefGoogle Scholar
  4. Chang L, Karin M (2001) Mammalian MAP kinase signalling cascades. Nature 410(6824):37–40CrossRefGoogle Scholar
  5. Chen H, Wu D, Ding X, Ying W (2015) SIRT2 is required for lipopolysaccharide-induced activation of BV2 microglia. Neuroreport 26(2):88–93. CrossRefPubMedGoogle Scholar
  6. Das A, Kim SH, Arifuzzaman S, Yoon T, Chai JC, Lee YS, Park KS, Jung KH, Chai YG (2016) Transcriptome sequencing reveals that LPS-triggered transcriptional responses in established microglia BV2 cell lines are poorly representative of primary microglia. J Neuroinflammation 13(1):182. CrossRefPubMedPubMedCentralGoogle Scholar
  7. Dong C, Davis RJ, Flavell RA (2002) MAP kinases in the immune response. Annu Rev Immunol 20:55–72CrossRefGoogle Scholar
  8. Gomes P, Fleming Outeiro T, Cavadas C (2015) Emerging role of Sirtuin 2 in the regulation of mammalian metabolism. Trends Pharmacol Sci 36(11):756–768. CrossRefPubMedGoogle Scholar
  9. Hammer M, Mages J, Dietrich H, Servatius A, Howells N, Cato AC, Lang R (2006) Dual specificity phosphatase 1 (DUSP1) regulates a subset of LPS-induced genes and protects mice from lethal endotoxin shock. J Exp Med 203(1):15–20CrossRefGoogle Scholar
  10. Harrison IF, Smith AD, Dexter DT (2018) Pathological histone acetylation in Parkinson’s disease: neuroprotection and inhibition of microglial activation through SIRT 2 inhibition. Neurosci Lett 666:48–57. CrossRefPubMedPubMedCentralGoogle Scholar
  11. Henn A, Lund S, Hedtjärn M, Schrattenholz A, Pörzgen P, Leist M (2009) The suitability of BV2 cells as alternative model system for primary microglia cultures or for animal experiments examining brain inflammation. ALTEX 26(2):83–94CrossRefGoogle Scholar
  12. Hickman S, Izzy S, Sen P, Morsett L, El Khoury J (2018) Microglia in neurodegeneration. Nat Neurosci 21(10):1359–1369. CrossRefPubMedPubMedCentralGoogle Scholar
  13. Hoogland IC, Houbolt C, van Westerloo DJ, van Gool WA, van de Beek D (2015) Systemic inflammation and microglial activation: systematic review of animal experiments. J Neuroinflammation 12:114. CrossRefPubMedPubMedCentralGoogle Scholar
  14. Jung YJ, Lee AS, Nguyen-Thanh T, Kim D, Kang KP, Lee S, Park SK, Kim W (2015) SIRT2 regulates LPS-induced renal tubular CXCL2 and CCL2 expression. J Am Soc Nephrol 26(7):1549–1560. CrossRefPubMedGoogle Scholar
  15. Kim HS, Vassilopoulos A, Wang RH, Lahusen T, Xiao Z, Xu X, Li C, Veenstra TD, Li B, Yu H, Ji J, Wang XW, Park SH, Cha YI, Gius D, Deng CX (2011) SIRT2 maintains genome integrity and suppresses tumorigenesis through regulating APC/C activity. Cancer Cell 20(4):487–499. CrossRefPubMedPubMedCentralGoogle Scholar
  16. Korhonen R, Moilanen E (2014) Mitogen-activated protein kinase phosphatase 1 as an inflammatory factor and drug target. Basic Clin Pharmacol Toxicol 114(1):24–36. CrossRefPubMedGoogle Scholar
  17. Lawan A, Shi H, Gatzke F, Bennett AM (2013) Diversity and specificity of the mitogen-activated protein kinase phosphatase-1 functions. Cell Mol Life Sci 70(2):223–237. CrossRefPubMedGoogle Scholar
  18. Lee AS, Jung YJ, Kim D, Nguyen-Thanh T, Kang KP, Lee S, Park SK, Kim W (2014) SIRT2 ameliorates lipopolysaccharide-induced inflammation in macrophages. Biochem Biophys Res Commun 450(4):1363–1369. CrossRefPubMedGoogle Scholar
  19. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods 25(4):402–408CrossRefGoogle Scholar
  20. Mendes KL, Lelis DF, Santos SHS (2017) Nuclear sirtuins and inflammatory signaling pathways. Cytokine Growth Factor Rev 38:98–105. CrossRefPubMedGoogle Scholar
  21. North BJ, Marshall BL, Borra MT, Denu JM, Verdin E (2003) The human Sir2 ortholog, SIRT2, is an NAD+-dependent tubulin deacetylase. Mol Cell 11(2):437–444CrossRefGoogle Scholar
  22. Ory D, Celen S, Verbruggen A, Bormans G (2014) PET radioligands for in vivo visualization of neuroinflammation. Curr Pharm Des 20(37):5897–5913CrossRefGoogle Scholar
  23. Pais TF, Szegő ÉM, Marques O, Miller-Fleming L, Antas P, Guerreiro P, de Oliveira RM, Kasapoglu B, Outeiro TF (2013) The NAD-dependent deacetylase sirtuin 2 is a suppressor of microglial activation and brain inflammation. EMBO J 32(19):2603–2616. CrossRefPubMedPubMedCentralGoogle Scholar
  24. Rahimifard M, Maqbool F, Moeini-Nodeh S, Niaz K, Abdollahi M, Braidy N, Nabavi SM, Nabavi SF (2017) Targeting the TLR4 signaling pathway by polyphenols: a novel therapeutic strategy for neuroinflammation. Ageing Res Rev 36:11–19. CrossRefPubMedGoogle Scholar
  25. Schwartz M, Deczkowska A (2016 Oct) Neurological disease as a failure of brain-immune crosstalk: the multiple faces of Neuroinflammation. Trends Immunol 37(10):668–679. CrossRefPubMedGoogle Scholar
  26. Serrano L, Martínez-Redondo P, Marazuela-Duque A, Vazquez BN, Dooley SJ, Voigt P, Beck DB, Kane-Goldsmith N, Tong Q, Rabanal RM, Fondevila D, Muñoz P, Krüger M, Tischfield JA, Vaquero A (2013) The tumor suppressor SirT2 regulates cell cycle progression and genome stability by modulating the mitotic deposition of H4K20 methylation. Genes Dev 27(6):639–653. CrossRefPubMedPubMedCentralGoogle Scholar
  27. Song GJ, Suk K (2017) Pharmacological modulation of functional phenotypes of microglia in neurodegenerative diseases. Front Aging Neurosci 9:139. CrossRefPubMedPubMedCentralGoogle Scholar
  28. Suzuki T, Khan MN, Sawada H, Imai E, Itoh Y, Yamatsuta K, Tokuda N, Takeuchi J, Seko T, Nakagawa H, Miyata N (2012) Design, synthesis, and biological activity of a novel series of human sirtuin-2-selective inhibitors. J Med Chem 55(12):5760–5773. CrossRefPubMedGoogle Scholar
  29. Wang B, Zhang Y, Cao W, Wei X, Chen J, Ying W (2016) SIRT2 plays significant roles in lipopolysaccharides-induced Neuroinflammation and brain injury in mice. Neurochem Res 41(9):2490–2500. CrossRefPubMedGoogle Scholar
  30. Wang J, Koh HW, Zhou L, Bae UJ, Lee HS, Bang IH, Ka SO, Oh SH, Bae EJ, Park BH (2017) Sirtuin 2 aggravates postischemic liver injury by deacetylating mitogen-activated protein kinase phosphatase-1. Hepatology 65(1):225–236. CrossRefPubMedGoogle Scholar
  31. Xanthos DN, Sandkühler J (2014) Neurogenic neuroinflammation: inflammatory CNS reactions in response to neuronal activity. Nat Rev Neurosci 15(1):43–53. CrossRefPubMedGoogle Scholar
  32. Xie XQ, Zhang P, Tian B, Chen XQ (2017) Downregulation of NAD-dependent Deacetylase SIRT2 protects mouse brain against ischemic stroke. Mol Neurobiol 54(9):7251–7261. CrossRefPubMedGoogle Scholar
  33. Zhao Q, Wang X, Nelin LD, Yao Y, Matta R, Manson ME, Baliga RS, Meng X, Smith CV, Bauer JA, Chang CH, Liu Y (2006) MAP kinase phosphatase 1 controls innate immune responses and suppresses endotoxic shock. J Exp Med 203(1):131–140CrossRefGoogle Scholar
  34. Zhao T, Alam HB, Liu B, Bronson RT, Nikolian VC, Wu E, Chong W, Li Y (2015) Selective inhibition of SIRT2 improves outcomes in a lethal septic model. Curr Mol Med 15(7):634–641CrossRefGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Infectious DiseasesRenmin Hospital of Wuhan UniversityWuhanChina

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