S100A9通过TLR4/NF-κB信号通路活化星形胶质细胞促进炎症反应

周鑫; 白巧; 张小印; 叶柳; 彭彦; 唐勇; 刘永刚

doi:10.13417/j.gab.041.001103

S100A9通过TLR4/NF-κB信号通路活化星形胶质细胞促进炎症反应

周鑫¹,
白巧¹,
张小印¹,
叶柳²,
彭彦¹,
唐勇¹,
刘永刚^1, ,

1. 重庆医科大学基础医学院干细胞与组织工程研究室;
2. 重庆医科大学附属第一医院生殖医学中心

基金项目:

重庆市前沿与应用基础研究项目(cstc2015jcyjA10034)资助

详细信息

通讯作者:
刘永刚,liu.yonggang@cqmu.edu.cn

中图分类号: R741.02
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出版历程
- 刊出日期: 2022-05-24

S100A9 Activates Astrocytes Through TLR4/NF-κB Signaling Pathway to Promote Inflammation

1. Laboratory of Stem Cells and Tissue Engineering, College of Basic Medicine, Chongqing Medical University;
2. Reproductive Medicine Center, The First Affliated Hospital of Chongqing Medical University

摘要

摘要: 为探讨S100A9诱导星形胶质细胞活化及促炎机制，本研究采用不同浓度S100A9蛋白作用于小鼠星形胶质细胞24 h, CCK-8检测星形胶质细胞增殖；划痕实验检测细胞迁移能力；RT-qPCR检测胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)和补体C3 mRNA表达水平；ELISA检测细胞上清液中白细胞介素-6(interleukin-6, IL-6)和肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)蛋白水平；细胞转录组测序筛选出TLR4/NF-κB信号通路；Western blot检测细胞中TLR4、MYD88、P65和PP65蛋白含量；TLR4/NF-κB信号通路的活化情况验证。结果表明，与对照组相比，5μg/mL S100A9对星形胶质细胞有明显的促进增殖和迁移的作用；S100A9可以显著上调活化标志物GFAP、C3以及炎症因子IL-6、TNF-α,并上调细胞表达TLR4、MYD88、P65和PP65蛋白水平；应用TLR4受体抑制剂TAK-242和NF-κB通路抑制剂BAY11-7082可以显著逆转星形胶质细胞活化及炎症因子的分泌。综上所述，S100A9促进星形胶质细胞的增殖和迁移，通过TLR4/NF-κB信号通路促进星形胶质细胞A1型活化及分泌炎症因子促进炎症反应。
- S100A9 /
- 星形胶质细胞 /
- 炎症 /
- NF-κB /
- TLR4
Abstract: To investigate the mechanism of S100 A9 inducing astrocyte activation and promoting inflammation, mouse astrocytes were treated with different concentrations of S100 A9 protein for 24 h. CCK-8 was used to detect the astrocyte proliferation rate; scratch test was applied to detect the migration ability; the mRNA levels of glial fibrillary acidic protein（GFAP） and complement C3 were tested by RT-qPCR; the protein levels of IL-6 and TNF-α were detected by ELISA in the culture supernatant; the signaling pathway was screened by transcriptional sequencing; the protein level of TLR4, MyD88, P65 and PP65 were evaluated by Western blot; the activation of TLR4/NF-κB signaling pathway was verified. The results showed that 5 μg/mL S100 A9 promoted the proliferation and migration of astrocytes compared to the control group; S100 A9 significantly up-re-gulated the protein level of IL-6, TNF-α, GFAP, C3, TLR4, MYD88, P65 and PP65; addition of TAK-242 and BAY11-7082, the inhibitors of TLR4 and NF-κB pathway respectively, can significantly reverse the activation of astrocytes and the production of inflammatory factors. In summary, S100 A9 promotes A1 astrocyte activation through activating TLR4/NF-κB signaling pathway to enhance inflammatory response.
- S100A9 /
- Astrocyte /
- Inflammations /
- NF-κB /
- TLR4

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参考文献(18)

[1]	Carter S.F.,Herholz K.,Rosa-Neto P.,Pellerin L.,Nordberg A.,and Zimmer E.R.,2019,Astrocyte biomar-kers in Alzheimer′s disease,Trends Mol.Med.,25(2):77-95.
[2]	Escartin C.,Galea E.,Lakatos A.,O′Callaghan P.,Petzold G.C.,Serrano-Pozo A.,Steinhäuser C.,Vol-terra A.,Carmignoto G.,Agarwal A.,Allen N.J.,Araque A.,Barbeito L.,Barzilai A.,Bergles D.E.,Bonvento G.,Butt A.M.,Chen W.T.,Cohen-Salmon M.,Cunningham C.,Deneen B.,De Strooper B.,Diaz-Castro B.,Farina C.,Freeman M.,Gallo V.,Goldman J.E.,Goldman S.A.,Gotz M.,Gutierrez A.,Haydon P.G.,Heiland D.H.,Hol E.M.,Holt M.G.,Iino M.,Kastanenka K.V.,Kettenmann H.,Khakh B.S.,Koizumi S.,Lee C.J.,Liddelow S.A.,MacVicar B.A.,Magistretti P.,Messing A.,Mishra A.,Molofsky A.V.,Murai K.K.,Norris C.M.,Okada S.,Oliet S.H.R.,Oliveira J.F.,Panatier A.,Parpura V.,Pekna M.,Pekny M.,Pellerin L.,Perea G.,Perez-Nievas B.G.,Pfrieger F.W.,Poskanzer K.E.,Quintana F.J.,Ransohoff R.M.,Riquelme-Perez M.,Robel S.,Rose C.R.,Rothstein J.D.,Rouach N.,Rowitch D.H.,Semyanov A.,Sirko S.,Sontheimer H.,Swanson R.A.,Vitorica J.,Wanner I.B.,Wood L.B.,Wu J.,Zheng B.,Zimmer E.R.,Zorec R.,Sofroniew M.V.,and Verkhratsky A.,2021,Reactive astrocyte nomenclature,definitions,and future directions,Nat.Neurosci.,24(3):312-325.
[3]	Horvath I.,Jia X.,Johansson P.,Wang C.,Moskalenko R.,Steinau A.,Forsgren L.,Wagberg T.,Svensson J.,Zetterberg H.,and Morozova-Roche L.A.,2016,Pro-inflammatory S100A9 protein as a robust biomar-ker differentiating early stages of cognitive impairment in Alzheimer′s disease,ACS Chem.Neurosci.,7(1):34-39.
[4]	Kim H.J.,Chang K.A.,Ha T.Y.,Kim J.,Ha S.,Shin K.Y.,Moon C.,Nacken W.,Kim H.S.,and Suh Y.H.,2014,S100A9 knockout decreases the memory impairment and neuropathology in crossbreed mice of Tg2576 and S100A9 knockout mice model,PLoS ONE,9(2):e88924.
[5]	Kwon H.S.,and Koh S.H.,2020,Neuroinflammation in neurodegenerative disorders:the roles of microglia and astrocytes,Transl.Neurodegener.,9(1):42.
[6]	Liao J.M.,Ye L.,Huang Q.,Tian Y.,Hu K.,Li X.P.,Tang Y.,and Liu Y.G.,2018,Experimental study on the effect of S100A9 on the apoptosis of astrocytes in SD rats in vitro and its mechanism,Genomics and Applied Biology,37(8):3652-3659.(廖键梅,叶柳,黄琴,田银,胡凯,李晓朋,唐勇,刘永刚,2018,S100A9促进体外SD大鼠星形胶质细胞凋亡及其机制的实验研究,基因组学与应用生物学,37(8):3652-3659.)
[7]	Liddelow S.A.,and Barres B.A.,2017,Reactive astrocytes:production,function,and therapeutic potential,Immunity,46(6):957-967.
[8]	Liddelow S.A.,Guttenplan K.A.,Clarke L.E.,Bennett F.C.,Bohlen C.J.,Schirmer L.,Bennett M.L.,Münch A.E.,Chung W.S.,Peterson T.C.,Wilton D.K.,Frouin A.,Napier B.A.,Panicker N.,Kumar M.,Buckwalter M.S.,Rowitch D.H.,Dawson V.L.,Dawson T.M.,Stevens B.,and Barres B.A.,2017,Neurotoxic reactive astrocytes are induced by activated microglia,Nature,541(7638):481-487.
[9]	Li L.,Acioglu C.,Heary R.F.,and Elkabes S.,2021,Role of astroglial toll-like receptors (TLRs) in central nervous system infections,injury and neurodegenerative diseases,Brain Behav.Immun.,91:740-755.
[10]	Liu H.X.,Wu X.,Luo J.N.,Wang X.G.,Guo H.,Feng D.Y.,Zhao L.,Bai H.,Song M.Y.,Liu X.Y.,Guo W.,Li X.,Yue L.,Wang B.D.,and Qu Y.,2019,Pterostilbene attenuates astrocytic inflammation and neuronal oxidative injury after ischemia-reperfusion by inhibiting NF-κB phosphorylation,Front.Immunol.,10:2408.
[11]	Linnerbauer M.,Wheeler M.A.,and Quintana F.J.,2020,Astrocyte crosstalk in CNS inflammation,Neuron,108(4):608-622.
[12]	Pereira C.F.,Santos A.E.,Moreira P.I.,Pereira A.C.,Sousa F.J.,Cardoso S.M.,and Cruz M.T.,2019,Is Alzheimer′s disease an inflammasomopathy,Ageing Research Reviews,56:100966.
[13]	Ryu K.Y.,Lee H.J.,Woo H.,Kang R.J.,Han K.M.,Park H.,Lee S.M.,Lee J.Y.,Jeong Y.J.,Nam H.W.,Nam Y.,and Hoe H.S.,2019,Dasatinib regulates LPS-induced microglial and astrocytic neuroinflammatory responses by inhibiting AKT/STAT3 signaling,J.Neuroinflammation,16(1):190.
[14]	Sofroniew M.V.,2020,Astrocyte reactivity:subtypes,states,and functions in CNS innate immunity,Trends Immunol.,41(9):758-770.
[15]	Toleikis Z.,Ziaunys M.,Baranauskiene L.,Petrauskas V.,Jaudzems K.,and Smirnovas V.,2021,S100A9 alters the pathway of alpha-synuclein amyloid aggregation,Int.J.Mol.Sci.,22(15):7972.
[16]	Urso D.,Gnoni V.,Filardi M.,and Logroscino G.,2021,Delusion and delirium in neurodegenerative disorders:an overlooked relationship?Front.Psychiatry,12:808724.
[17]	Wang C.,Iashchishyn I.A.,Pansieri J.,Nyström S.,Klementieva O.,Kara J.,Horvath I.,Moskalenko R.,Rofougaran R.,Gouras G.,Kovacs G.G.,Shankar S.K.,and Morozova-Roche L.A.,2018,S100A9-dri-ven amyloid-neuroinflammatory cascade in traumatic brain injury as a precursor state for Alzheimer′s di-sease,Sci.Rep.,8(1):12836.
[18]	Wu M.,Xu L.,Wang Y.,Zhou N.,Zhen F.,Zhang Y.,Qu X.B.,Fan H.B.,Liu S.H.,Chen Y.,and Yao R.Q.,2018,S100A8/A9 induces microglia activation and promotes the apoptosis of oligodendrocyte precursor cells by activating the NF-κB signaling pathway,Brain Res.Bull.,143:234-245.