IL-4在哮喘中通过增加自噬通路和线粒体氧化应激来激活ULK1/Atg9a/Rab9、NLRP3炎症小体和高尔基体碎裂
2024/02/23
摘要
在哮喘发作期间，肺上皮炎症、线粒体氧化应激和高尔基体碎裂现象加剧。然而，其潜在机制仍然大部分未知。因此，本研究调查了ULK1、Atg9a和Rab9在上皮炎症、线粒体氧化应激和高尔基体碎裂中的作用。我们发现ULK1基因敲除减少了炎症细胞的浸润，恢复了Th1/Th2比例的失衡，并抑制了屋尘螨诱导的哮喘小鼠肺组织中炎症小体的形成。此外，我们证明了Atg9a与ULK1在S467位点相互作用。ULK1在S14位点磷酸化了Atg9a。使用ULK1激活剂（LYN-1604）和ULK1抑制剂（ULK-101）分别促进和抑制炎症小体的活化，表明屋尘螨诱导的哮喘小鼠中炎症小体的活化依赖于ULK1。为了验证体内结果，我们使用含有ULK1野生型和ULK1-S467A基因的慢病毒感染Beas-2b-ULK1敲除细胞并建立了稳定的细胞系。结果表明，ULK1的S467位点对IL-4诱导的炎症和氧化应激至关重要。实验证实，Atg9a是Rab9的优先信号通路。有趣的是，我们首次发现Rab9在炎症诱导的高尔基体碎裂中发挥了非常重要的作用。抑制ULK1/Atg9a/Rab9信号通路的激活可以抑制哮喘中的高尔基体碎裂和线粒体氧化应激，同时减少NLRP3介导的肺上皮炎症的产生。
IL-4 activates ULK1/Atg9a/Rab9 in asthma, NLRP3 inflammasomes, and Golgi fragmentation by increasing autophagy flux and mitochondrial oxidative stress
Chang Xu, Yilan Song, Wanting Liu, Ruobai Liu, Qiaoyun Bai, Liangchang Li, Chongyang Wang, Guanghai Yan
Abstract
During asthma, there is an intensification of pulmonary epithelial inflammation, mitochondrial oxidative stress, and Golgi apparatus fragmentation. However, the underlying mechanism remains largely unknown. Therefore, this study investigated the roles of ULK1, Atg9a, and Rab9 in epithelial inflammation, mitochondrial oxidative stress, and Golgi apparatus fragmentation. We found that ULK1 gene knockout reduced the infiltration of inflammatory cells, restored the imbalance of the Th1/Th2 ratio, and inhibited the formation of inflammatory bodies in the lung tissue of house dust mite-induced asthma mice. Moreover, we demonstrated that Atg9a interacted with ULK1 at S467. ULK1 phosphorylated Atg9a at S14. Treatment with ULK1 activator (LYN-1604) and ULK1 inhibitor (ULK-101) respectively promoted and inhibited inflammasome activation, indicating that the activation of inflammasome induced by house dust mite in asthma mice is dependent on ULK1. For validation of the in vivo results, we then used a lentivirus containing ULK1 wild type and ULK1-S467A genes to infect Beas-2b-ULK1-knockout cells and establish a stable cell line. The results suggest that the ULK1 S467 site is crucial for IL-4-induced inflammation and oxidative stress. Experimental verification confirmed that Atg9a was the superior signaling pathway of Rab9. Interestingly, we found for the first time that Rab9 played a very important role in inflammation-induced fragmentation of the Golgi apparatus. Inhibiting the activation of the ULK1/Atg9a/Rab9 signaling pathways can inhibit Golgi apparatus fragmentation and mitochondrial oxidative stress in asthma while reducing the production of NLRP3-mediated pulmonary epithelial inflammation.
在哮喘发作期间，肺上皮炎症、线粒体氧化应激和高尔基体碎裂现象加剧。然而，其潜在机制仍然大部分未知。因此，本研究调查了ULK1、Atg9a和Rab9在上皮炎症、线粒体氧化应激和高尔基体碎裂中的作用。我们发现ULK1基因敲除减少了炎症细胞的浸润，恢复了Th1/Th2比例的失衡，并抑制了屋尘螨诱导的哮喘小鼠肺组织中炎症小体的形成。此外，我们证明了Atg9a与ULK1在S467位点相互作用。ULK1在S14位点磷酸化了Atg9a。使用ULK1激活剂（LYN-1604）和ULK1抑制剂（ULK-101）分别促进和抑制炎症小体的活化，表明屋尘螨诱导的哮喘小鼠中炎症小体的活化依赖于ULK1。为了验证体内结果，我们使用含有ULK1野生型和ULK1-S467A基因的慢病毒感染Beas-2b-ULK1敲除细胞并建立了稳定的细胞系。结果表明，ULK1的S467位点对IL-4诱导的炎症和氧化应激至关重要。实验证实，Atg9a是Rab9的优先信号通路。有趣的是，我们首次发现Rab9在炎症诱导的高尔基体碎裂中发挥了非常重要的作用。抑制ULK1/Atg9a/Rab9信号通路的激活可以抑制哮喘中的高尔基体碎裂和线粒体氧化应激，同时减少NLRP3介导的肺上皮炎症的产生。
(中日友好医院呼吸与危重症医学科 顾宪民 摘译 林江涛 审校)
(Redox Biol. 2024 Feb 15:71:103090. doi: 10.1016/j.redox.2024.103090.)
(Redox Biol. 2024 Feb 15:71:103090. doi: 10.1016/j.redox.2024.103090.)
IL-4 activates ULK1/Atg9a/Rab9 in asthma, NLRP3 inflammasomes, and Golgi fragmentation by increasing autophagy flux and mitochondrial oxidative stress
Chang Xu, Yilan Song, Wanting Liu, Ruobai Liu, Qiaoyun Bai, Liangchang Li, Chongyang Wang, Guanghai Yan
Abstract
During asthma, there is an intensification of pulmonary epithelial inflammation, mitochondrial oxidative stress, and Golgi apparatus fragmentation. However, the underlying mechanism remains largely unknown. Therefore, this study investigated the roles of ULK1, Atg9a, and Rab9 in epithelial inflammation, mitochondrial oxidative stress, and Golgi apparatus fragmentation. We found that ULK1 gene knockout reduced the infiltration of inflammatory cells, restored the imbalance of the Th1/Th2 ratio, and inhibited the formation of inflammatory bodies in the lung tissue of house dust mite-induced asthma mice. Moreover, we demonstrated that Atg9a interacted with ULK1 at S467. ULK1 phosphorylated Atg9a at S14. Treatment with ULK1 activator (LYN-1604) and ULK1 inhibitor (ULK-101) respectively promoted and inhibited inflammasome activation, indicating that the activation of inflammasome induced by house dust mite in asthma mice is dependent on ULK1. For validation of the in vivo results, we then used a lentivirus containing ULK1 wild type and ULK1-S467A genes to infect Beas-2b-ULK1-knockout cells and establish a stable cell line. The results suggest that the ULK1 S467 site is crucial for IL-4-induced inflammation and oxidative stress. Experimental verification confirmed that Atg9a was the superior signaling pathway of Rab9. Interestingly, we found for the first time that Rab9 played a very important role in inflammation-induced fragmentation of the Golgi apparatus. Inhibiting the activation of the ULK1/Atg9a/Rab9 signaling pathways can inhibit Golgi apparatus fragmentation and mitochondrial oxidative stress in asthma while reducing the production of NLRP3-mediated pulmonary epithelial inflammation.
上一篇：
塑料添加剂暴露诱导的炎症性脂质和脂肪酸代谢异常在儿童哮喘中的作用
下一篇：
哮喘患者中气道上皮细胞对RSV的反应主要表现为粘液细胞和多纤毛细胞的受损
