Identification and validation of metformin protects against PM2.5-induced macrophages cytotoxicity by targeting toll like receptor pathway.
Identification and validation of metformin protects against PM2.5-induced macrophages cytotoxicity by targeting toll like receptor pathway.
复制标题
DOI：
10.1016/j.chemosphere.2020.126526
复制
发表时间：
2020-07
期刊：
影响因子：
8.8
通讯作者：
Yanfeng Shi;Josevata Werelagi Batibawa;Modibo Maiga;Baiyang Sun;Yang Li;Junchao Duan;Zhiwei Sun
中科院分区：
文献类型：
--
作者：
Yanfeng Shi;Josevata Werelagi Batibawa;Modibo Maiga;Baiyang Sun;Yang Li;Junchao Duan;Zhiwei Sun
细颗粒物（PM2.5）已被广泛报道与肺部疾病的发病机制有关。最近有报道称二甲双胍可减轻pm2.5相关的呼吸和心血管损伤，但其潜在机制尚未发现。本文利用GEO数据库中的RNAseq图谱，对二甲双胍的保护作用及其机制进行了全面的生物信息学分析和充分的验证实验。结合edgeR、主成分分析（PCA）、K-Means聚类、基因集富集分析（GSEA）、GO和KEGG富集等多种生物信息学工具，鉴定了TLRs/MyD88/NF-κB轴在pm2.5相关毒性过程中起关键信号转导作用。pm2.5激活TLRs/MyD88/NF-κB通路，导致RAW264.7细胞IL-6、TNF-α的显著产生，线粒体损伤，细胞活力降低，LDH活性升高。二甲双胍通过限制TLRs/MyD88/NF-κB通路，显著降低IL-6的产生、线粒体损伤、细胞活力和LDH活性。将抗AMPKα2或阴性对照的siRNA转染RAW264.7细胞，以鉴定二甲双胍是否以依赖AMPKα2的方式保护pm2.5诱导的细胞毒性。二甲双胍预处理显著减弱pm2.5诱导的siAMPKα2细胞活力降低、LDH活性升高、TLRs/MyD88/NF-κB通路抑制。综上所述，我们的研究结果表明，二甲双胍通过以不依赖AMPKα2的方式抑制TLRs/MyD88/NF-κB信号通路，保护pm2.5诱导的线粒体损伤和细胞毒性。
Fine particle matter (PM2.5) has been extensively reported to contribute to the pathogenesis of pulmonary diseases. Recently, metformin has been reported to attenuate PM2.5associated respiratory and cardiovascular injury, but the underling mechanism has not been discovered. Here, we performed comprehensively bioinformatics analysis and fully validation experiment to investigate the protection role of metformin and underling mechanism with RNAseq profile in GEO database. A combination of various bioinformatics tools including edgeR, principal component analysis (PCA), K-Means clustering, Gene Set Enrichment Analysis (GSEA), GO and KEGG enrichment were performed to identify the TLRs/MyD88/NF-κB axis functional as the key signaling transduction during PM2.5associated toxicity. PM2.5activated TLRs/MyD88/NF-κB pathway and resulted in significantly generation of IL-6, TNF-α, mitochondrial damage, decreasing of cell viability and increased LDH activity in RAW264.7 cells. Metformin significantly attenuated the production of IL-6, mitochondrial damage, cell viability and LDH activity by limiting TLRs/MyD88/NF-κB pathway. The siRNA against AMPKα2 or negative control were transfected to RAW264.7 cells to identify whether metformin protects PM2.5-induced cytotoxicity in an AMPKα2-dependent manner. Pretreatment with metformin significantly attenuated PM2.5induced decreasing of cell viability and increased LDH activity, as well as inhibited the TLRs/MyD88/NF-κB pathway in both siControl or siAMPKα2 cells. Taken together, our results indicate that metformin protects against PM2.5-induced mitochondrial damage and cell cytotoxicity by inhibiting TLRs/MyD88/NF-κB signaling pathway in an AMPKα2 independent manner.