KLF10通过下调NF-κB/NLRP3通路改善大鼠焦虑性抑郁行为及神经炎症OA
KLF10 overexpression alleviates neuroinflammation and anxiety-depressive disorder in rats by downregulating the NF-κB/NLRP3 pathway
目的 解析昼夜节律基因KLF10作为焦虑性抑郁生物标志物的神经免疫调控机制.方法 基于GEO数据库人源外周血基因芯片信息,纳入健康对照64例、抑郁组62例、焦虑性抑郁59例,结合MSigDB数据库筛选差异表达的昼夜节律基因.加权基因共表达网络分析和机器学习模型筛选核心基因,KEGG通路富集分析生物过程,采用ROC曲线评估诊断效能.将40只SPF级雄性SD大鼠按体质量均质化分为空白组、假手术组、模型组、KLF10过表达组,10只/组,通过脑立体定位-侧脑室注射腺相关病毒(AAV)构建KLF10过表达模型、慢性束缚联合皮质酮应激构建焦虑性抑郁大鼠模型.强迫游泳试验检测抑郁样行为,高架十字迷宫检测焦虑样行为;采用尼氏染色观察海马、杏仁核病理学改变,酶联免疫吸附法(ELISA)检测中枢及外周细胞因子白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、白介素-10(IL-10)、转化生长因子-β(TGF-β);免疫荧光染色观察KLF10与p-p65共定位表达;Western blotting及qRT-PCR检测NF-κB/NLRP3通路分子表达水平.结果 抑郁患者较健康人群9个昼夜节律基因存在表达差异,焦虑性抑郁与单纯抑郁相比免疫浸润评分升高,KEGG涉及NOD样受体信号通路和NF-κB信号通路的上调.特异性生物标记物KLF10的诊断效能为0.885(P<0.001).在体实验验证表明,焦虑性抑郁模型大鼠强迫游泳不动时间增加、高架十字迷宫开臂探索减少(P<0.05),中枢及外周促炎因子IL-1β、TNF-α含量升高,抗炎因子IL-10、TGF-β含量下降(P<0.05);海马和杏仁核神经元排列紊乱、尼氏体减少.NF-κB/NLRP3通路各信号分子转录水平及关键蛋白表达升高(P<0.05).过表达KLF10可显著改善大鼠抑郁样及焦虑样行为(P<0.05),恢复中枢和外周促炎/抗炎因子平衡,改善海马及杏仁核神经元排列,同时上调NF-κB/NLRP3通路抑制节点NFKBIA的转录(P<0.001),从而下调NLRP3、RELA的转录(P<0.05),降低p-IκB-α、p-p65、NLRP3蛋白的表达(P<0.05).结论 KLF10通过下调NF-κB/NLRP3通路抑制海马-杏仁核炎症,改善抑郁及焦虑行为,可作为焦虑性抑郁诊断标志物与治疗靶点.
Objective To elucidate the neuroimmune regulatory mechanism of the circadian rhythm gene KLF10 as a biomarker for anxiety depressive disorder(ADD).Methods The differentially expressed circadian rhythm genes were screened using human peripheral blood gene chip data from the GEO database(including 64 healthy individuals,62 patients with major depressive disorder[MDD],and 59 with ADD)in conjunction with the MSigDB database.Weighted gene co-expression network analysis and machine learning models were employed to identify the core genes,followed by KEGG pathway enrichment analysis and evaluation of their diagnostic efficacy using ROC curves.In a male SD rat model of ADD induced by chronic restraint and corticosterone stress,the changes in depressive-like behaviors,hippocampal and amygdala pathologies,levels of inflammatory and pro-inflammatory cytokines,co-localization of KLF10 and p-p65 expression,and expression levels of NF-κB/NLRP3 pathway molecules were examined following stereotactic AAV virus injection into the lateral ventricle for KLF10 overexpression.Results Compared with healthy individuals,the depressive patients showed differential expressions of 9 circadian rhythm genes.Compared with the MDD patients,the patients with ADD had significantly higher immune infiltration scores with upregulated NOD-like receptor and NF-κB signaling pathways.The specific biomarker KLF10 demonstrated a diagnostic efficacy of 0.885.In the rat models of AOD,KLF10 overexpression significantly ameliorated depression-and anxiety-like behaviors,restored the balance between the pro-and anti-inflammatory cytokines,and improved hippocampal and amygdala pathologies.KLF10 overexpression also markedly upregulated NFKBIA mRNA,downregulated NLRP3 and RELA mRNAs,and reduced protein expressions of p-IκB-α,p-p65,and NLRP3 in the brain tissues of the rats.Conclusion KLF10 overexpression ameliorates ADD behaviors in rats by inhibiting hippocampal-amygdala inflammation via downregulating the NF-κB/NLRP3 pathway,suggesting the potential of KLF10 as a diagnostic biomarker and therapeutic target for AOD.
刘安澜;过伟峰;李建香;张天鸽;徐丹
广州中医药大学基础医学院,广东 广州 510006||南京中医药大学第一临床医学院,江苏 南京 210023南京中医药大学第一临床医学院,江苏 南京 210023南京中医药大学第一临床医学院,江苏 南京 210023广州中医药大学基础医学院,广东 广州 510006南京中医药大学第一临床医学院,江苏 南京 210023||南京中医药大学附属太仓医院(太仓市中医医院)脑病科,江苏 苏州 215400
焦虑性抑郁神经炎症NF-κB/NLRP3通路昼夜节律基因
anxiety-depressive disorderneuro-inflammationNF-κB/NLRP3 pathwaycircadian rhythm genes
《南方医科大学学报》 2026 (5)
994-1005,12
国家自然科学基金(82074309)江苏省教育厅江苏省研究生科研创新计划项目(KYCX23_2123)苏州市科技局基础应用研究(SKYD2023236)Supported by National Natural Science Foundation of China(82074309).
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