1 Program in Cell Biology, Department of Pediatrics, and.
Am J Respir Cell Mol Biol. 2014 Apr;50(4):825-37. doi: 10.1165/rcmb.2013-0386OC.
Idiopathic pulmonary fibrosis (IPF) is a relentless, fibrotic parenchymal lung disease in which alternatively programmed macrophages produce profibrotic molecules that promote myofibroblast survival and collagen synthesis. Effective therapies to treat patients with IPF are lacking, and conventional therapy may be harmful. We tested the hypothesis that therapeutic lung delivery of the proinflammatory cytokine tumor necrosis factor (TNF)-α into wild-type fibrotic mice would reduce the profibrotic milieu and accelerate the resolution of established pulmonary fibrosis. Fibrosis was assessed in bleomycin-instilled wild-type and TNF-α(-/-) mice by measuring hydroxyproline levels, static compliance, and Masson's trichrome staining. Macrophage infiltration and programming status was assessed by flow cytometry of enzymatically digested lung and in situ immunostaining. Pulmonary delivery of TNF-α to wild-type mice with established pulmonary fibrosis was found to reduce their fibrotic burden, to improve lung function and architecture, and to reduce the number and programming status of profibrotic alternatively programmed macrophages. In contrast, fibrosis and alternative macrophage programming were prolonged in bleomycin-instilled TNF-α(-/-) mice. To address the role of the reduced numbers of alternatively programmed macrophages in the TNF-α-induced resolution of established pulmonary fibrosis, we conditionally depleted macrophages in MAFIA (MAcrophage Fas-Induced Apoptosis) mice. Conditional macrophage depletion phenocopied the resolution of established pulmonary fibrosis observed after therapeutic TNF-α delivery. Taken together, our results show for the first time that TNF-α is involved in the resolution of established pulmonary fibrosis via a mechanism involving reduced numbers and programming status of profibrotic macrophages. We speculate that pulmonary delivery of TNF-α or augmenting its signaling pathway represent a novel therapeutic strategy to resolve established pulmonary fibrosis.
特发性肺纤维化(IPF)是一种进行性、纤维化的肺实质疾病,其中替代性编程的巨噬细胞产生促纤维化分子,促进肌成纤维细胞的存活和胶原合成。缺乏治疗 IPF 患者的有效疗法,常规治疗可能有害。我们测试了这样一个假设,即向野生型纤维化小鼠的肺部输送促炎细胞因子肿瘤坏死因子(TNF)-α,可减少促纤维化环境并加速已建立的肺纤维化的消退。通过测量羟脯氨酸水平、静态顺应性和 Masson 三色染色,评估博莱霉素诱导的野生型和 TNF-α(-/-)小鼠的纤维化。通过酶消化肺和原位免疫染色的流式细胞术评估巨噬细胞浸润和编程状态。向已建立肺纤维化的野生型小鼠肺部输送 TNF-α 可降低其纤维化负担,改善肺功能和结构,并减少促纤维化的替代性编程巨噬细胞的数量和编程状态。相比之下,博莱霉素诱导的 TNF-α(-/-)小鼠的纤维化和替代性巨噬细胞编程延长。为了解决 TNF-α诱导的已建立肺纤维化消退中替代性编程巨噬细胞数量减少的作用,我们在 MAFIA(巨噬细胞 Fas 诱导凋亡)小鼠中条件性耗尽巨噬细胞。条件性巨噬细胞耗竭模仿了在治疗性 TNF-α给药后观察到的已建立肺纤维化的消退。总之,我们的研究结果首次表明,TNF-α通过涉及减少促纤维化巨噬细胞的数量和编程状态的机制参与已建立的肺纤维化的消退。我们推测,肺部输送 TNF-α或增强其信号通路代表了一种治疗已建立的肺纤维化的新策略。