摘要
目的探讨Zunyimycin C调控小胶质细胞的分子机制及其抗阿尔茨海默病(Alzheimer's disease,AD)的潜在价值。方法构建Aβ25-35诱导的BV2小胶质细胞炎症模型,整合LC-MS/MS蛋白质组学技术解析差异表达蛋白(differentially expressed proteins,DEP),结合KEGG通路富集分析与RT-qPCR验证,系统揭示Zunyimycin C对MAPK/ERK、NF-κB通路及炎症因子(iNOS、Arg-1等)的调控机制,并通过透射电镜观察线粒体超微结构动态变化。结果模型组与对照组相比,共鉴定出188个DEP(P<0.05,|log2FC|≥1),其中122个上调、66个下调。高剂量Zunyimycin C(2.5µmol/L)显著逆转80个DEP的表达,包括下调炎症相关蛋白Lamtor5(-1.8倍)、Hspb1(-1.5倍)和Rom1(-1.7倍)。KEGG分析显示,Zunyimycin C主要富集于MAPK信号通路(P<0.01)、铁死亡通路(P<0.01)及干细胞多能性调控通路(P<0.05)。机制研究证实,Zunyimycin C通过抑制ERK磷酸化(p-ERK1/2减少42%)和NFκB核转位(p65核蛋白减少38%),显著下调促炎因子iNOS(-1.9倍)和CD86(-1.6倍),同时上调抗炎因子Arg-1(+1.3倍)和CD206(+1.5倍)。电镜结果显示,Zunyimycin C干预可有效逆转模型组细胞形态改变,减少线粒体/溶酶体数量,促进自噬小体形成。结论Zunyimycin C通过MAPK/PTEN/PI3K通路交叉调控实现小胶质细胞炎症状态的动态平衡,其抗炎效果优于阳性对照姜黄素,为AD的治疗提供了新策略。
Objective To explore the molecular mechanisms of Zunyimycin C in regulating microglial cells and its potential against Alzheimer's disease(AD).Methods In this study,an Aβ25-35-induced BV2 microglial cell model was used.LC-MS/MS proteomics analyzed differentially expressed proteins(DEP)after Zunyimycin C treatment.KEGG pathway enrichment and RT-qPCR verified changes in key pathways(MAPK/ERK,NF-κB)and inflammatory factors(iNOS,Arg-1,etc.).Transmission electron microscopy assessed mitochondrial ultrastructural changes and cellular and organelle alterations.Results Compared to the control group,188 DEP were identified in the model group(P<0.05,|log2FC|≥1),with 122 upregulated and 66 downregulated.High-dose Zunyimycin C(2.5µmol/L)significantly reversed the expression of 80 DEP,including downregulating inflammation-related proteins Lamtor5(-1.8×),Hspb1(-1.5×),and Rom1(-1.7×).KEGG analysis revealed Zunyimycin C enrichment in MAPK signaling(P<0.01),ferroptosis(P<0.01),and stem cell pluripotency regulation(P<0.05).Mechanistic studies confirmed that Zunyimycin C inhibited ERK phosphorylation(p-ERK1/2 reduced by 42%)and NF-κB nuclear translocation(p65 nuclear protein reduced by 38%),downregulated pro-inflammatory factors iNOS(-1.9×)and CD86(-1.6×),and upregulated anti-inflammatory factors Arg-1(+1.3×)and CD206(+1.5×).Electron microscopy revealed that,compared to the normal group,the model group exhibited activated inflammatory cell conditions,with notable changes in cell shape,membrane wrinkles with protrusions,more pseudopodia,and increased mitochondria and lysosomes.After Zunyimycin C intervention,these pro-inflammatory changes were suppressed.Cells elongated,with more endoplasmic reticulum and Golgi apparatus,and autophagic changes like autophagosomes and mitophagy appeared.Conclusion Zunyimycin C dynamically balances microglial inflammation via multi-target synergy(MAPK/PTEN/PI3K pathway crosstalk),showing better anti-inflammatory effects than curcumin,offering a new AD intervention strategy.
作者
林丹
杨甜
梁厶爻
王卓灵
郑敏
李倩
吕玉红
LIN Dan;YANG Tian;LIANG Siyao;WANG Zhuoling;ZHENG Min;LI Qian;LYU Yuhong(Yan'an Medical College of Yan'an University,Yan'an 716000,China;Yan'an Key Laboratory of Microbial Drug Innovation and Transformation,Yan'an 716000,China;Department of Laboratory Medicine,Yan'an University Affiliated Hospital,Yan'an 716000,China.)
出处
《延安大学学报(医学科学版)》
2025年第2期1-9,共9页
Journal of Yan'an University:Medical Science Edition
基金
国家自然科学基金(82060654)。
