ACT001 suppressing M1 polarization against inflammation via NF-?B and STAT1 signaling pathways alleviates acute lung injury in mice
ACT001 suppressing M1 polarization against inflammation via NF-?B and STAT1 signaling pathways alleviates acute lung injury in mice
复制标题
DOI：
10.1016/j.intimp.2022.108944
复制
发表时间：
2022-09-01
影响因子：
5.6
通讯作者：
Chen, Zhe
中科院分区：
文献类型：
--
作者：
Guo, Hui;Song, Yan;Chen, Zhe
ACT 001已显示出优异的抗肿瘤和抗纤维化活性。然而，ACT 001在急性肺损伤（ALI）中的作用及其潜在机制仍不清楚。本研究旨在探讨ACT 001对急性肺损伤的保护作用及其机制。本研究首次建立了脂多糖（LPS）腹腔注射致小鼠急性肺损伤模型。ACT 001治疗显著减轻了ALI小鼠肺组织的组织病理学变化，肺M1巨噬细胞浸润减少。然后，我们进行了体外实验，发现ACT 001处理有效地抑制了RAW 264.7和THP-1的M1表型。接下来，我们进行了下拉和质谱分析，以筛选ACT 001的相互作用蛋白，确定IKK β和STAT 1为ACT 001的关键靶蛋白。ACT 001处理显著抑制NF-κ B和STAT 1通路，从而抑制体内和体外M1对抗炎症的极化。最后，我们使用IMD 0354（IMD）和氟达拉滨（Flud）特异性阻断IKK β和STAT 1的活性，并通过IKK β和STAT 1过表达刺激巨噬细胞。我们的数据清楚地表明，ACT 001诱导的M1极化降低被IMD和Flud处理阻断，并被RAW 264.7细胞中IKK β和STAT 1过表达逆转。总之，我们发现ACT 001通过抑制NF-κ B B和STAT 1信号通路，显著减轻炎症反应，限制肺巨噬细胞的M1表型，为开发治疗ALI/ARDS的药物提供了新的思路。
ACT001 has been shown to exhibit excellent antitumor and anti-fibrosis activities. However, the role of ACT001 in acute lung injury (ALI) and the underlying mechanism remains largely unclear. The present study aimed to investigate the protective effects of ACT001 on ALI and explore the potential mechanisms. Herein, we firstly established the ALI mouse model induced by intratracheal instillation of lipopolysaccharide (LPS). ACT001 treatment significantly alleviated histopathological changes of lung tissues with lower infiltration of pulmonary M1 macrophages in ALI mice. Then, we performed in vitro experiment and found that ACT001 treatment effectively inhibited the M1 phenotype of RAW264.7 and THP-1.. Next, we performed pull-down and mass spectrometry analysis to screen the interacting proteins of ACT001, identifying IKK beta and STAT1 as the critical target proteins of ACT001. And ACT001 treatment significantly suppressed the NF-Kappa B and STAT1 pathways, thereby inhibiting the M1 polarization against inflammation in vivo and in vitro. Finally, we used IMD 0354 (IMD) and Fludarabine (Flud) to specifically block the activity of IKK beta and STAT1, and stimulated macrophages through IKK beta and STAT1 overexpression. Our data clearly showed that ACT001-induced decrease of the M1 polarization was blocked by IMD and Flud treatment, and reversed by IKK beta and STAT1 overexpression in RAW264.7 cells. In conclusion, we discovered that ACT001 significantly alleviates inflammation and limits M1 phenotype of pulmonary macrophages via suppressing NF-Kappa B and STAT1 signaling pathways, providing new insights for the development of drugs to treat ALI/ARDS.