Macrophage Notch1 Participates in LPS-Induced Acute Lung Injury via Regulating CCR5 Expression in Mice.

IF 3.3 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Ruiyu Zhang, Hui Du, Zhi Liu, Yuxi Lei, Huizhi Hu, Junwen Zheng, Pu Yang, Dongchi Zhao
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Abstract

Background: As pivotal immunoregulatory sentinels in pulmonary defense systems, alveolar macrophages (AMs) play dual roles in mediating inflammatory responses and tissue repair processes during various phases of inflammatory cascades. The present investigation focuses on elucidating the regulatory influence of Notch pathway activation within AM populations on the pathophysiological mechanisms underlying acute lung injury (ALI) development.

Methods: To investigate the regulatory roles of Notch intracellular domain (NICD) and C-C chemokine receptor type 5 (CCR5) in pulmonary inflammation, an ALI model was established through lipopolysaccharide (LPS) administration. Complementary studies used macrophage-specific Notch1 knockout mice and immortalized bone marrow-derived macrophages (iBMDMs). Molecular profiling of CCR5 and inflammatory mediators was performed through real-time quantitative reverse transcription PCR (qRT-PCR) and immunofluorescence staining. Functional assessments of macrophage migration were carried out using scratch wound healing assays and transwell migration assays.

Results: In the LPS-induced ALI model, pulmonary tissues exhibited elevated expression of both NICD and CCR5. Conversely, Notch1 knockout mice attenuated CCR5 expression, reduced macrophage infiltration and downregulated transcription of pro-inflammatory mediators compared to wild-type controls (p < 0.05). Lung injury was milder in the Notch1-deficient mice model compared to wild mice (p < 0.05). In vitro experiments demonstrated that inhibiting the Notch pathway in macrophages reduced CCR5 expression and attenuated CCL5-induced macrophage migration.

Conclusion: Notch signaling regulates macrophage infiltration and the inflammatory response by modulating CCR5 expression in ALI induced by LPS.

巨噬细胞Notch1通过调节CCR5表达参与lps诱导的小鼠急性肺损伤
背景:肺泡巨噬细胞(alveolar macrophages, AMs)作为肺防御系统中关键的免疫调节哨兵,在炎症级联反应的各个阶段介导炎症反应和组织修复过程中发挥双重作用。本研究的重点是阐明AM群体中Notch通路激活对急性肺损伤(ALI)发生的病理生理机制的调节作用。方法:采用脂多糖(LPS)诱导ALI模型,研究Notch胞内结构域(NICD)和C-C趋化因子受体5型(CCR5)在肺部炎症中的调节作用。补充研究使用巨噬细胞特异性Notch1敲除小鼠和永生化骨髓源性巨噬细胞(iBMDMs)。通过实时定量反转录PCR (qRT-PCR)和免疫荧光染色对CCR5和炎症介质进行分子谱分析。通过抓伤愈合试验和跨井迁移试验对巨噬细胞迁移进行功能评估。结果:在lps诱导的ALI模型中,肺组织中NICD和CCR5的表达均升高。相反,与野生型对照组相比,Notch1敲除小鼠CCR5表达减弱,巨噬细胞浸润减少,促炎介质转录下调(p < 0.05)。notch1缺失小鼠模型肺损伤较野生小鼠轻(p < 0.05)。体外实验表明,抑制巨噬细胞Notch通路可降低CCR5的表达,减轻ccl5诱导的巨噬细胞迁移。结论:Notch信号通过调节LPS诱导ALI中CCR5的表达,调控巨噬细胞浸润及炎症反应。
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