Acta NaturaeActa NaturaeActa Naturae2075-8251Acta Naturae Ltd1183010.32607/actanaturae.11830Research ArticlesЭкспериментальные статьиResearch ArticleAttenuating Neuronal Autophagy Alleviates Inflammatory Injury in OGD-Deprived Co-culture of HT22 with BV2Attenuating Neuronal Autophagy Alleviates Inflammatory Injury in OGD-Deprived Co-culture of HT22 with BV2https://orcid.org/0000-0002-4023-1431HuangZhiwen
China
h386434161@live.comLiuYuyuan
China
h386434161@live.comChenXuemei
China
h386434161@live.comYuChunlei
China
yuchunlei1828820@163.comHeHongyun
China
511869321@qq.comDengYihao
China
h386434161@live.comKunming University of Science and TechnologyAnning First People’s Hospital Affiliated to Kunming University of Science and Technology3010202315391991010202224072023Copyright ©; 2023, Huang Z., Liu Y., Chen X., Yu C., He H., Deng Y.Copyright ©; 2023, Huang Z., Liu Y., Chen X., Yu C., He H., Deng Y.2023Huang Z., Liu Y., Chen X., Yu C., He H., Deng Y.Huang Z., Liu Y., Chen X., Yu C., He H., Deng Y.https://creativecommons.org/licenses/by/4.0https://actanaturae.ru/2075-8251/article/view/11830

Neuronal CX3CL1 suppressed microglial inflammation by binding to its receptor CX3CR1 expressed on microglia. Neuronal autophagy was prominently activated by cerebral ischemia, whereas CX3CL1 expression in autophagic neurons was conversely down-regulated to exacerbate microglial inflammation. Accordingly, this study was meant to investigate whether ischemia-activated microglial inflammation could be repressed by promoting CX3CL1 expression via the attenuation of neuronal autophagy. Immunofluorescence showed that autophagy predominantly occurred in neurons but barely in microglia. Western blot and immunofluorescence demonstrated that attenuating HT22 autophagy significantly increased its CX3CL1 expression and subsequently mitigated the BV2-mediated inflammatory responses, as indicated by decreased inflammatory factors of NF-κB-p65, IL-6, IL-1β, TNF-α, and PGE2. Meanwhile, CCK-8, Nissl staining, and FJC staining showed that an OGD (Oxygen-glycogen deprivation)-created neuronal injury was greatly alleviated by CX3CL1-suppressed microglial inflammation. Contrarily, elevating HT22 autophagy markedly decreased its CX3CL1 expression, which consequently worsened microglial inflammation and the neuronal injury. Our data suggests that attenuating neuronal autophagy may be an effective method to alleviate a microglial inflammatory injury after an ischemic stroke.

Neuronal CX3CL1 suppressed microglial inflammation by binding to its receptor CX3CR1 expressed on microglia. Neuronal autophagy was prominently activated by cerebral ischemia, whereas CX3CL1 expression in autophagic neurons was conversely down-regulated to exacerbate microglial inflammation. Accordingly, this study was meant to investigate whether ischemia-activated microglial inflammation could be repressed by promoting CX3CL1 expression via the attenuation of neuronal autophagy. Immunofluorescence showed that autophagy predominantly occurred in neurons but barely in microglia. Western blot and immunofluorescence demonstrated that attenuating HT22 autophagy significantly increased its CX3CL1 expression and subsequently mitigated the BV2-mediated inflammatory responses, as indicated by decreased inflammatory factors of NF-κB-p65, IL-6, IL-1β, TNF-α, and PGE2. Meanwhile, CCK-8, Nissl staining, and FJC staining showed that an OGD (Oxygen-glycogen deprivation)-created neuronal injury was greatly alleviated by CX3CL1-suppressed microglial inflammation. Contrarily, elevating HT22 autophagy markedly decreased its CX3CL1 expression, which consequently worsened microglial inflammation and the neuronal injury. Our data suggests that attenuating neuronal autophagy may be an effective method to alleviate a microglial inflammatory injury after an ischemic stroke.

ischemic strokeneuronal autophagyCX3CL1 expressionmicroglial inflammationneuroprotectionischemic strokeneuronal autophagyCX3CL1 expressionmicroglial inflammationneuroprotectionNational Natural Science Foundation of ChinaNos.81960418, 82160240, 82160241Yunnan Ten Thousand Talents Plan Young & Elite Talents ProjectNo. YNWR-QNBJ-2018-034Yunnan Applied Basic Research Projects Fund of Yunnan Provincial Department of Science & TechnologyNo.202001AT070049Wang P., Shao B.Z., Deng Z., Chen S., Yue Z., Miao C.Y. // Prog. Neurobiol. 2018. V. 163. P. 98–117.Iadecola C., Anrather J. // Nat. Med. 2011. V. 17. № 7. P. 796–808.Su P., Zhang J., Wang D., Zhao F., Cao Z., Aschner M., Luo W. // Neuroscience. 2016. V. 319. P. 155–167.Berglund R., Guerreiro-Cacais A.O., Adzemovic M.Z., Zeitelhofer M., Lund H., Ewing E., Ruhrmann S., Nutma E., Parsa R., Thessen-Hedreul M., et al. // Sci. Immunol. 2020. V. 5. № 52. P. 5077.Lu M., Zhao W., Han S., Lin X., Xu T., Tan Q., Wang M., Yi C., Chu X., Yang W., et al. // Sci. Adv. 2022. V. 8. № 26. P. 8048.Mendiola A.S., Church K.A., Cardona S.M., Vanegas D., Garcia S.A., Macklin W., Lira S.A., Ransohoff R.M., Kokovay E., Lin C-HA., et al. // J. Neurochem. 2022. V. 162. № 5. P. 430–443.Chen X., Jiang M., Li H., Wang Y., Shen H., Li X., Zhang Y., Wu J., Yu Z., Chen G. // J. Neuroinflammation. 2020. V. 17. № 1. P. 209–224.Lauro C., Chece G., Monaco L., Antonangeli F., Peruzzi G., Rinaldo S., Paone A., Cutruzzolà F., Limatola C. // Front. Cell Neurosci. 2019. V. 13. P. 414.Xu S., Lu J., Shao A., Zhang J.H., Zhang J. // Front. Immunol. 2020. V. 11. P. 294.Subbarayan M.S., Joly-Amado A., Bickford P.C., Nash K.R. // Pharmacol. Ther. 2022. V. 231 P. 107989.Liu M., Xu Z., Wang L., Zhang L., Liu Y., Cao J., Fu Q., Liu Y., Li H., Lou J., et al. // J. Neuroinflammation. 2020. V. 17. № 1. P. 270.Campbell B.C.V., Khatri P. // Lancet. 2020. V. 396. № 10244. P. 129–142.Zhang Y., Wu Z., Huang Z., Liu Y., Chen X., Zhao X., He H., Deng Y. // Brain Res. 2022. V. 1778. P. 147768.Camargos Q.M., Silva B.C., Silva D.G., Toscano E.Cd.B., Oliveira B.d.S., Bellozi P.M.Q., Jardim B.L.d.O., Vieira É.L.M., de Oliveira A.C.P., Sousa L.P., et al. // Brain Res. Bull. 2020. V. 155. P. 1–10.Ermine C.M., Bivard A., Parsons M.W., Baron J.-C. // Int. J. Stroke. 2021. V. 16. № 5. P. 497509.He H.Y., Ren L., Guo T., Deng Y.H. // Neural. Regen. Res. 2019. V. 14. № 2. P. 280–288.Wu Z., Zhang Y., Liu Y., Chen X., Huang Z., Zhao X., He H., Deng Y. // Life (Basel). 2021. V. 11. № 9. P. 948.Ajoolabady A., Wang S., Kroemer G., Penninger J.M., Uversky V.N., Pratico D., Henninger N., Reiter R.J., Bruno A., Joshipura K., et al. // Pharmacol. Ther. 2021. V. 225. P. 107848.Barthels D., Das H. // Biochim. Biophys. Acta Mol. Basis Dis. 2020. V. 1866. № 4. P. 165260.Al-Mufti F., Amuluru K., Roth W., Nuoman R., El-Ghanem M., Meyers P.M. // Neurosurgery. 2018. V. 82. № 6. P. 781–789.Yu S., Yu M., Bu Z., He P., Feng J. // Front. Cell Neurosci. 2020. V. 14. P. 193.Lingling D., Miaomiao Q., Yili L., Hongyun H., Yihao D. // Brain Res. Bull. 2022. V. 184. P. 24–33.Lian L., Zhang Y., Liu L., Yang L., Cai Y., Zhang J., Xu S. // Front. Mol. Neurosci. 2020. V. 13. P. 612439.Cipriani R., Villa P., Chece G., Lauro C., Paladini A., Micotti E., Perego C., De Simoni M.G., Fredholm B.B., Eusebi F., et al. // J. Neurosci. 2011. V. 31. № 45. P. 16327–16335.Mao M., Xu Y., Zhang X.Y., Yang L., An X.B., Qu Y., Chai Y.N., Wang Y.R., Li T.T., Ai J. // J. Neuroinflammation. 2020. V. 17. № 1. P. 244.