肝细胞生长因子通过激活NF-κB信号通路促进BV2细胞介导神经炎症  

Hepatocyte growth factor promotes neuroinflammation mediated by BV2 cells through activating NF-κB signaling pathway

作　　者：沈若琪 鲁玉宝 王子铭 刘斌[1] 张良明[1] 杨阳[1] Shen Ruoqi;Lu Yubao;Wang Ziming;Liu Bin;Zhang Liangming;Yang Yang(Department of Spine Surgery,the Third Affiliated Hospital of Sun Yat-sen University,Guangzhou 510000,Guangdong Province,China)

机构地区：[1]中山大学附属第三医院脊柱外科,广州510000

出　　处：《中国临床解剖学杂志》2024年第6期666-672,共7页Chinese Journal of Clinical Anatomy

基　　金：广东省基础与应用基础研究基金项目(2023A1515012473);国家自然科学基金项目(82372400);国家自然科学基金培育项目(2023GZRPYMS04)。

摘　　要：目的 探讨肝细胞生长因子(hepatocyte growth factor,HGF)对小胶质细胞介导神经炎症的作用及其调控机制,为后续可能治疗靶点的确定提供实验依据和理论参考。方法 选取小鼠小胶质细胞(BV2)作为研究对象,分为对照组和HGF组,通过蛋白质组学分析、实时荧光定量聚合酶链反应(qRTPCR)以及蛋白质印迹法(western blot,WB)等检测HGF对BV2细胞NF-κB信号通路活性及炎症因子表达的影响。结果 蛋白质组学分析显示HGF处理后,NF-κB通路显著富集;HGF处理1h时,BV2细胞增殖活力随浓度升高下降,6h和12h时呈现先上升后下降的趋势;qRT-PCR显示HGF显著上调炎症因子(iNOS、TNF-α、IL-1β);WB表明HGF促进炎症相关因子(TNF-α、NF-κB p65)相对表达量上升。结论本研究首次证实HGF通过激活BV2细胞NF-κB信号通路促进神经炎症的作用及其机制,为进一步探究神经炎症调控策略及开发有效的临床治疗手段提供重要线索。Objective To explore the effect of hepatocyte growth factor(HGF)on microglia-mediated neuroinflammation and its regulatory mechanism,and to provide experimental basis and theoretical reference for the determination of possible treatment targets.Methods The mouse immortalized cell line-BV2 cells,which retain multiple morphological and functional characteristics of microglia,were divided into a control group and a HGF group.The effects of HGF on the activity of NF-κB signaling pathway and subsequent expression of neuroinflammatory factors were determined by tyrosine phosphorylated proteomic analysis,cell survival assay,quantitative real time polymerase chain reaction(qRT-PCR)and western blot(WB).Results Tyrosine phosphorylated proteomic analysis showed that after addition of HGF,NF-κB signaling was one of the most enriched pathways in BV2 cells.At 1 hour after the addition of HGF with different concentrations,the proliferation activity of BV2 cells was decreased.At both 6 hours and 12 hours after the supplementation of HGF with various concentrations,the proliferation activity of BV2 cells increased at early stage,while decreased at late stage.qRT-PCR found that HGF could upregulate the transcription level of inflammatory factors derived from BV2 cells(iNOS,TNF-α,IL-1β).WB also showed that HGF could promote protein translation of associated inflammatory factors(TNF-α,NF-κB p65).Conclusions This study confirms the role of HGF in promoting neuroinflammation by activating the NF-κB signaling pathway in BV2 cells and its mechanism.It provides important clues to further explore the regulatory strategies on neuroinflammation to develop more effective treatment in the clinic.

关 键 词：肝细胞生长因子 小胶质细胞 神经炎症 NF-ΚB信号通路 因子 

分 类 号：R745.4[医药卫生—神经病学与精神病学]