NLRP3 Inflammasome Activation Contributes to Mechanical Stretch-Induced Endothelial-Mesenchymal Transition and Pulmonary Fibrosis
NLRP3 Inflammasome Activation Contributes to Mechanical Stretch-Induced Endothelial-Mesenchymal Transition and Pulmonary Fibrosis
复制标题
NLRP3 炎症小体激活有助于机械拉伸诱导的内皮-间质转化和肺纤维化
DOI：
10.1097/ccm.0000000000002799
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发表时间：
2018-01-01
影响因子：
8.8
通讯作者：
Zhu, Xiao-Yan
中科院分区：
文献类型：
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作者：
Lv, Zhou;Wang, Yan;Zhu, Xiao-Yan
目的：机械通气可诱发肺纤维化。本研究旨在探讨呼吸机诱导的肺纤维化是否与内皮-间质转化相关，并揭示其潜在机制。设计：随机对照动物实验和细胞培养实验。环境：大学研究实验室。研究对象：成年男性肿瘤研究所NACHT、LRR、PYD结构域含蛋白3 （NLRP3）敲除及野生型小鼠。原代培养小鼠肺血管内皮细胞。干预措施：肿瘤研究所、NLRP3敲除小鼠和野生型小鼠给予机械通气（20mL/kg） 2小时。小鼠肺血管内皮细胞循环拉伸24小时。测量结果及主要结果：机械通气小鼠肺组织中胶原沉积、羟脯氨酸和I型胶原含量、转化生长因子- β 1含量均增加。通气诱导的肺纤维化与间充质标志物（α -平滑肌肌动蛋白和vimentin）的表达增加以及内皮标志物（血管内皮-钙粘蛋白和CD31）的表达降低有关。双免疫荧光染色显示CD31/ α平滑肌肌动蛋白、CD31/vimentin和CD31/成纤维细胞特异性蛋白-1在肺组织中共定位，提示内皮-间质转化形成。机械通气也可诱导肺组织NLRP3炎性体活化。在体外直接机械拉伸原代小鼠肺血管内皮细胞可导致类似的NLRP3激活和内皮-间质转化形成，而NLRP3敲低可阻止这一过程。此外，与野生型小鼠相比，nlrp3缺陷小鼠的机械拉伸诱导的内皮-间质转化和肺纤维化得到改善。结论：机械拉伸可能通过nlrp3依赖途径促进内皮-间质转化和肺纤维化。通过NLRP3失活抑制内皮-间质转化可能是对抗机械通气相关肺纤维化的可行治疗策略。
Objectives: Mechanical ventilation can induce lung fibrosis. This study aimed to investigate whether ventilator-induced lung fibrosis was associated with endothelial-mesenchymal transition and to uncover the underlying mechanisms.Design: Randomized, controlled animal study and cell culture study.Setting: University research laboratory.Subjects: Adult male Institute of Cancer Research, NACHT, LRR, and PYD domains-containing protein 3 (NLRP3) knockout and wild-type mice. Primary cultured mouse lung vascular endothelial cells.Interventions: Institute of Cancer Research, NLRP3 knockout and wild-type mice were subjected to mechanical ventilation (20mL/kg) for 2 hours. Mouse lung vascular endothelial cells were subjected to cyclic stretch for 24 hours.Measurements and Main Results: Mice subjected to mechanical ventilation exhibited increases in collagen deposition, hydroxyproline and type I collagen contents, and transforming growth factor-beta 1 in lung tissues. Ventilation-induced lung fibrosis was associated with increased expression of mesenchymal markers (alpha smooth muscle actin and vimentin), as well as decreased expression of endothelial markers (vascular endothelial-cadherin and CD31). Double immunofluorescence staining showed the colocalization of CD31/alpha smooth muscle actin, CD31/vimentin, and CD31/fibroblast-specific protein-1 in lung tissues, indicating endothelial-mesenchymal transition formation. Mechanical ventilation also induced NLRP3 inflammasome activation in lung tissues. In vitro direct mechanical stretch of primary mouse lung vascular endothelial cells resulted in similar NLRP3 activation and endothelial-mesenchymal transition formation, which were prevented by NLRP3 knockdown. Furthermore, mechanical stretch-induced endothelial-mesenchymal transition and pulmonary fibrosis were ameliorated in NLRP3-deficient mice as compared to wild-type littermates.Conclusions: Mechanical stretch may promote endothelial-mesenchymal transition and pulmonary fibrosis through a NLRP3-dependent pathway. The inhibition of endothelial-mesenchymal transition by NLRP3 inactivation may be a viable therapeutic strategy against pulmonary fibrosis associated with mechanical ventilation.