线粒体自噬调控脊髓损伤的机制及潜在治疗靶点OA
Mechanisms and potential therapeutic targets of mitophagy regulation in spinal cord injury
目的:研究线粒体自噬在脊髓损伤(Spinal cord injury,SCI)治疗中的作用机制与潜在靶点,以明确其治疗潜力,从而为改善SCI预后提供新策略.方法:从GEO数据库中小鼠脊髓数据集GSE5296和GSE47681获取表达谱数据,用于识别SCI与线粒体自噬相关基因间的差异表达基因(Differentially expressed genes,DEGs).通过通路富集分析揭示DEGs的功能特征,构建蛋白质-蛋白质相互作用网络,并系统解析SCI微环境中的免疫细胞浸润特征.采用受试者工作特征(Receiver operating characteristic,ROC)曲线分析DEGs在SCI诊断中的效能.进一步通过逆转录定量聚合酶链反应(Quantitative real-time polymerase chain reaction,qRT-PCR)技术验证关键差异表达基因的转录水平,并借助蛋白质印迹法(Western blot)检测相关蛋白的表达水平,以综合评估其生物学意义.结果:筛选出55个与SCI和线粒体自噬相关的DEGs,其富集的信号通路与炎症、凋亡和细胞存活密切相关.基于PPI网络筛选出6个关键基因(Ulk1、Atg14、Map1lc3b、Gabarap、Gabarapl2和Fundc1),qRT-PCR分析结果显示,Atg14、Map1lc3b和Gabarapl2表达下调,Gabarap表达上调.免疫浸润分析显示,脊髓损伤后,10种免疫细胞丰度显著改变,其中中性粒细胞与M0巨噬细胞呈显著负相关(r=-0.56,P<0.05),Gabarap与M0巨噬细胞呈显著正相关(r=0.58,P<0.05),Gabarapl2与M0巨噬细胞呈显著负相关(r=-0.56,P<0.05).Western blot结果显示,SCI后NF-κB和p-ERK表达增加.结论:通过验证关键基因并识别免疫细胞的相互作用,确定了线粒体自噬在SCI治疗中的潜在靶点,为后续开发有效的SCI治疗方案指明了方向.
Objective:To explore the mechanisms and potential targets of mitophagy in the treatment of spinal cord injury(SCI)to clarify its therapeutic potential,thereby providing novel strategies for improving the prognosis of SCI.Methods:Expression profiles were obtained from the Gene Expression Omnibus(GEO)mouse and spinal cord datasets GSE5296 and GSE47681 to identify differentially expressed genes(DEGs)between SCI and mitophagy-related genes.Analyzed DEGs for pathway enrichment,constructed protein-protein interaction networks,and analyzed immune cell infiltration patterns in SCI.Receiver operating characteristic(ROC)curve analysis was used to evaluate the diagnostic value of DEGs in SCI.Verified the expression levels of key DEGs with reverse transcription-quantitative polymerase chain reaction(q RT-PCR).Analyzed protein levels with western blotting to comprehensively evaluate its biological significance.Results:We identified 55 DEGs associated with SCI and mitophagy,with their enriched signaling pathways closely related to inflammation,apoptosis,and cell survival.Six key genes were identified(Ulk1,Atg14,Map1lc3b,Gabarap,Gabarapl2,and Fundc1).The qRT-PCR analysis results showed that the expression of Atg14,Map1lc3b,and Gabarapl2 was downregulated,while the expression of Gabarap was upregulated.Immune infiltration analysis revealed significant alterations in the abundance of 10 immune cell types after spinal cord injury.Among these,neutrophils showed a strong negative correlation with M0 macrophages(r=-0.56,P<0.05).Meanwhile,Gabarap was significantly positively correlated with M0 macrophages(r=0.58,P<0.05),while Gabarapl2 was significantly negatively correlated with M0 macrophages(r=-0.56,P<0.05).Western blot results indicated increased expression of NF-κB and p-ERK following SCI.Conclusion:By validating the hub genes and identifying immune cell interactions,the potential targets of mitochondrial autophagy in SCI treatment were identified,which pointed out the direction for the subsequent development of effective SCI treatment.
吴欣;卢枫;王力峰;张雨;邱嘉欣;韩乐;谢海玉
赣南医科大学第一临床医学院赣南医科大学第一附属医院麻醉手术中心||赣州市麻醉学重点实验室,江西 赣州 341000赣南医科大学第一附属医院麻醉手术中心||赣州市麻醉学重点实验室,江西 赣州 341000赣南医科大学第一临床医学院赣南医科大学第一临床医学院赣南医科大学第一临床医学院赣南医科大学第一附属医院麻醉手术中心||赣州市麻醉学重点实验室,江西 赣州 341000
医药卫生
线粒体自噬脊髓损伤生物标志物免疫细胞浸润分子验证受试者工作特征曲线
Mitochondrial autophagySpinal cord injuryBiomarkersImmune cell infiltrationMolecular verificationReceiver operating characteristic curve
《赣南医科大学学报》 2026 (1)
16-25,10
江西省教育厅科学技术研究项目(编号:GJJ201505)
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