NETs通过TLR9信号通路促进M1巨噬细胞极化,从而加速主动脉瓣钙化。

IF 3.7 2区 生物学 Q3 CELL BIOLOGY
Luyao Liu, Zhenzhen Lan, Xincan Liu, Yun Chen, Zhihao Chen, Li Cheng, Ting Hu
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引用次数: 0

摘要

中性粒细胞胞外陷阱(NETs)与钙化主动脉瓣疾病(CAVD)的严重程度呈正相关。本研究旨在阐明net对CAVD的作用机制。通过钙化促进饮食建立CAVD小鼠模型,并通过腹腔注射氨基氯调节NETs的形成。我们在体外通过调节NETs和TLR9来观察NETs对Raw264.7细胞的影响。ELISA法检测TNF-α、MPO-DNA复合物、IL-10浓度。通过免疫荧光法观察瓜氨酸组蛋白H3 (citH3)的形成。采用qRT-PCR和Western blotting检测Raw264.7细胞BMP2、RUNX2、IL-1β、TNF-α、IL-10、tlr9的表达水平,流式细胞术检测CD86、CD206的表达水平。结果显示,与载药组相比,CAVD组瓣膜明显增厚,钙沉积增加,炎症因子TNF-α和IL-1β、net相关标志物MPO-DNA复合物和citH3、骨化因子BMP2和RUNX2、TLR9水平升高。相反,IL-10水平显著降低。cl -脒干预可显著改善早期CAVD小鼠瓣膜厚度,降低钙沉积、炎症因子、nets相关标志物、骨化因子和TLR9水平,同时增加IL-10水平。氯脒可能通过减少NETs来延缓小鼠CAVD的进展。体外研究证实,CAVD小鼠血清诱导NETs,通过TLR9信号通路促进Raw264.7细胞向M1表型极化,从而释放促炎因子(TNF-α、IL-1β、IL-6),抑制M2极化和IL-10表达。综上所述,我们的研究结果表明,NETs通过TLR9信号通路促进Raw264.7细胞向M1极化,促进了CAVD的炎症反应。本研究提出了一种新的针对NETs的治疗策略来延缓CAVD的进展。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
NETs accelerate aortic valve calcification by promoting M1 macrophage polarization through the TLR9 signaling pathway.

Neutrophil extracellular traps (NETs) are positively correlated with the severity of calcific aortic valve disease (CAVD). This study aims to elucidate the mechanism by which NETs contribute to CAVD. The CAVD mice model was established by calcification-promoting diets, and NETs formation was modulated via intraperitoneal injection of Cl-amidine. We observed the effect of NETs on Raw264.7 cells by regulating NETs and TLR9 in vitro. Concentrations of TNF-α, MPO-DNA complex, and IL-10 were measured using ELISA. NETs formation was assessed through immunofluorescence assay citrullinated histone H3 (citH3). Expression levels of BMP2, RUNX2, IL-1β, TNF-α, IL-10, and TLR 9 were analyzed by qRT-PCR and Western blotting, while flow cytometry was used to assess the expression of CD86 and CD206 on Raw264.7 cells. Results indicated that compared to the vehicle group, the CAVD group exhibited significant valve thickening and increased calcium deposition, as well as elevated levels of inflammatory factors TNF-α and IL-1β, NET-related markers MPO-DNA complexes and citH3, ossification factors BMP2 and RUNX2, and TLR9. Conversely, IL-10 levels were significantly reduced. Cl-amidine intervention in early CAVD mice significantly improved valve thickness and reduced calcium deposition, inflammatory factors, NETs-related markers, ossification factors, and TLR9 levels, while increasing IL-10 levels. Cl-amidine may delay CAVD progression in mice by reducing NETs. In vitro studies confirmed that serum from CAVD mice induced NETs, promoting the polarization of Raw264.7 cells to the M1 phenotype via TLR9 signaling pathway, thereby releasing pro-inflammatory factors (TNF-α, IL-1β, and IL-6), and inhibiting M2 polarization and IL-10 expression. In summary, our findings suggest that NETs promote Raw264.7 cell polarization to M1 through the TLR9 signaling pathway, contributing to the inflammatory response in CAVD. This study proposes a novel therapeutic strategy targeting NETs to delay CAVD progression.

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来源期刊
Molecular and Cellular Biochemistry
Molecular and Cellular Biochemistry 生物-细胞生物学
CiteScore
8.30
自引率
2.30%
发文量
293
审稿时长
1.7 months
期刊介绍: Molecular and Cellular Biochemistry: An International Journal for Chemical Biology in Health and Disease publishes original research papers and short communications in all areas of the biochemical sciences, emphasizing novel findings relevant to the biochemical basis of cellular function and disease processes, as well as the mechanics of action of hormones and chemical agents. Coverage includes membrane transport, receptor mechanism, immune response, secretory processes, and cytoskeletal function, as well as biochemical structure-function relationships in the cell. In addition to the reports of original research, the journal publishes state of the art reviews. Specific subjects covered by Molecular and Cellular Biochemistry include cellular metabolism, cellular pathophysiology, enzymology, ion transport, lipid biochemistry, membrane biochemistry, molecular biology, nuclear structure and function, and protein chemistry.
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