Roscioli Eugene, Tran Hai B, Jersmann Hubertus, Nguyen Phan T, Hopkins Emily, Lester Susan, Farrow Nigel, Zalewski Peter, Reynolds Paul N, Hodge Sandra
Lung Research Unit, Department of Thoracic Medicine, Hanson Institute, Royal Adelaide Hospital, Adelaide, South Australia, Australia;
Deptartment of Medicine, The University of Adelaide, Adelaide, South Australia, Australia.
Am J Physiol Lung Cell Mol Physiol. 2017 Sep 1;313(3):L453-L465. doi: 10.1152/ajplung.00083.2017. Epub 2017 Jun 8.
The proper regulation of zinc (Zn) trafficking proteins and the cellular distribution of Zn are critical for the maintenance of autophagic processes. However, there have been no studies that have examined Zn dyshomeostasis and the disease-related modulation of autophagy observed in the airways afflicted with chronic obstructive pulmonary disease (COPD). We hypothesized that dysregulated autophagy in airway epithelial cells (AECs) is related to Zn dysregulation in cigarette smoke (CS)-induced COPD. We applied a human ex vivo air-liquid interface model, a murine model of smoke exposure, and human lung tissues and investigated Zn, ZIP1, and ZIP2 Zn-influx proteins, autophagy [microtubule-associated 1A/1B-light chain-3 (LC3), Beclin-1], autophagic flux (Sequestosome), apoptosis [Bcl2; X-linked inhibitor of apoptosis (XIAP), poly (ADP)-ribose polymerase (PARP)], and inflammation [thymic stromal lymphopoietin (TSLP), regulated on activation, normal T cell expressed and secreted (RANTES), and IL-1β]. Lung tissues from CS-exposed mice exhibit reduced free-Zn in AECs, with elevated ZIP1 and diminished ZIP2 expression. Interestingly, increased LC3 colocalized with ZIP1, suggesting an autophagic requirement for free-Zn to support its catabolic function. In human AECs, autophagy was initiated but was unable to efficiently degrade cellular debris, as evidenced by stable Beclin-1 and increased LC3-II, but with a concomitant elevation in Sequestosome. Autophagic dysfunction due to CS exposure coupled with Zn depletion also induced apoptosis, with the reduction of antiapoptotic and antiautophagic proteins Bcl2 and XIAP and PARP cleavage. This was accompanied by an increase in RANTES and TSLP, an activator of adaptive immunity. We conclude that the uncoupling of Zn trafficking and autophagy in AECs constitutes a fundamental disease-related mechanism for COPD pathogenesis and could provide a new therapeutic target.
锌(Zn)转运蛋白的适当调节以及锌在细胞内的分布对于维持自噬过程至关重要。然而,目前尚无研究探讨慢性阻塞性肺疾病(COPD)气道中锌稳态失调以及与疾病相关的自噬调节情况。我们推测,香烟烟雾(CS)诱导的COPD中气道上皮细胞(AECs)自噬失调与锌调节异常有关。我们应用了人离体气液界面模型、烟雾暴露小鼠模型和人肺组织,研究了锌、ZIP1和ZIP2锌流入蛋白、自噬[微管相关蛋白1A/1B轻链3(LC3)、Beclin-1]、自噬通量(聚集体蛋白)、凋亡[Bcl2;X连锁凋亡抑制蛋白(XIAP)、聚(ADP-核糖)聚合酶(PARP)]以及炎症[胸腺基质淋巴细胞生成素(TSLP)、活化调节正常T细胞表达和分泌因子(RANTES)和IL-1β]。CS暴露小鼠的肺组织显示AECs中游离锌减少,ZIP1表达升高而ZIP2表达降低。有趣的是,增加的LC3与ZIP1共定位,表明自噬需要游离锌来支持其分解代谢功能。在人AECs中,自噬启动但无法有效降解细胞碎片,稳定的Beclin-1和增加的LC3-II证明了这一点,但同时聚集体蛋白也升高。CS暴露导致的锌缺乏引起的自噬功能障碍还诱导了细胞凋亡,抗凋亡和抗自噬蛋白Bcl2和XIAP减少以及PARP裂解。这伴随着RANTES和TSLP的增加,TSLP是适应性免疫的激活剂。我们得出结论,AECs中锌转运与自噬的解偶联构成了COPD发病机制中与疾病相关的基本机制,并可能提供一个新的治疗靶点。