干扰素调节因子3及其活化形式在hSOD1^(G93A)突变型肌萎缩侧索硬化症转基因模型中异常表达  

Abnormal expression of interferon regulatory factor 3 and its activated form in hSOD1^(G93A)mutant amyotrophic lateral sclerosis transgenic models

作　　者：高学帅 白雪 王晨晨 张雪 王雪枚 闫秋鹏 刘焕彩 陈燕春 Gao Xueshuai;Bai Xue;Wang Chenchen;Zhang Xue;Wang Xuemei;Yan Qiupeng;Liu Huancai;Chen Yanchun(Department of Histology and Embryology,and Shandong Province Key Laboratory of Neurological Diseases and Regenerative Repair,Shandong Second Medical University,Weifang 261053,China;Department of Joint Surgery,The Affiliated Hospital of Shandong Second Medical University,Weifang 261031,China)

机构地区：[1]山东第二医科大学基础医学院组织学与胚胎学教研室及神经疾病与再生修复重点实验室,潍坊261053 [2]山东第二医科大学附属医院关节外科,潍坊261031

出　　处：《中国组织化学与细胞化学杂志》2025年第1期9-17,共9页Chinese Journal of Histochemistry and Cytochemistry

基　　金：山东省自然科学基金(ZR2024MH112,ZR2024QH628);潍坊市卫生健康委员会科研项目(WFWSJK-2022-231)。

摘　　要：目的检测干扰素调节因子3(interferon regulatory factor 3,IRF3)及其活化形式(p-IRF3)在hSOD1^(G93A)突变型肌萎缩侧索硬化症(amyotrophic lateral sclerosis,ALS)转基因小鼠及细胞模型中的表达变化,为进一步阐明其与ALS发病的关系奠定基础。方法采用生物信息学分析方法筛选出IRF3为ALS差异表达基因,并选取hSOD1^(G93A)突变型ALS转基因小鼠、NSC34运动神经元样细胞模型、BV2细胞模型进行体内外实验验证。运用qRT-PCR、Western blot、免疫荧光染色技术检测不同发病阶段ALS转基因小鼠脊髓和大脑皮层中的IRF3及p-IRF3表达变化和定位,检测hSOD1^(G93A)突变型NSC34细胞和BV2细胞中IRF3及p-IRF3表达变化。结果生信分析显示,IRF3在ALS患者外周血单核细胞中表达异常上调,在ALS患者hiPSC衍生的运动神经元中也存在上调趋势;动物实验结果显示,与同窝WT小鼠相比,IRF3 mRNA以及IRF3、p-IRF3蛋白在ALS转基因小鼠发病期脊髓中表达上调,在大脑皮层中表达下调;在ALS小鼠脊髓前角检测到IRF3与NeuN、IRF3与GFAP以及IRF3与Iba1双标细胞。体外实验验证结果显示,IRF3在表达hSOD1^(G93A)突变型NSC34细胞和BV2细胞中表达上调。结论IRF3及其活化形式p-IRF3的异常表达与ALS发病相关。Objective To detect the expression changes of interferon regulatory factor 3(IRF3)and its activated form(p-IRF3)in hSOD1^(G93A)mutant amyotrophic lateral sclerosis(ALS)transgenic mice and cell models,and to lay a foundation for further clarifying their relationship with ALS pathogenesis.Methods Bioinformatics analysis was used to screen IRF3 as a differentially expressed gene in ALS.Transgenic mice,NSC34 motor neuron-like cells,and BV2 cells with hSOD1^(G93A)mutation were selected for in vivo and in vitro experiments.qRT-PCR,Western blot,and immunofluorescence staining were used to detect IRF3 and p-IRF3 expression and localization in the spinal cord and cerebral cortex of ALS transgenic mice at different disease stages,as well as in hSOD1^(G93A)mutant NSC34 and BV2 cells.Results Bioinformatics analysis showed abnormal upregulation of IRF3 in peripheral blood mononuclear cells of ALS patients,and an upward trend in hiPSC-derived motor neurons.Animal experiments found that compared with WT mice,IRF3 mRNA and IRF3 and p-IRF3 proteins were upregulated in the spinal cord of ALS transgenic mice during the disease stage,but downregulated in the cerebral cortex.IRF3 co-localized with NeuN,GFAP,and Iba1 in the ventral horn of the spinal cord.In vitro experiments confirmed IRF3 upregulation in hSOD1^(G93A)mutant NSC34 and BV2 cells.Conclusion Abnormal expression of IRF3 and p-IRF3 is associated with ALS pathogenesis.

关 键 词：肌萎缩侧索硬化症 脊髓 大脑皮层 干扰素调节因子3 p-IRF3 转基因小鼠 

分 类 号：R744.8[医药卫生—神经病学与精神病学]