Department of Physiology and Pennsylvania Muscle Institute, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, United States of America.
PLoS One. 2010 Oct 20;5(10):e13450. doi: 10.1371/journal.pone.0013450.
Duchenne's muscular dystrophy (DMD) is a severe progressive myopathy caused by mutations in the DMD gene leading to a deficiency of the dystrophin protein. Due to ongoing muscle necrosis in respiratory muscles late-stage DMD is associated with respiratory insufficiency and chronic hypoxia (CH). To understand the effects of CH on dystrophin-deficient muscle in vivo, we exposed the Drosophila model for DMD (dmDys) to CH during a 16-day ascent to the summit of Mount Denali/McKinley (6194 meters above sea level). Additionally, dmDys and wild type (WT) flies were also exposed to CH in laboratory simulations of high altitude hypoxia. Expression profiling was performed using Affymetrix GeneChips® and validated using qPCR. Hypoxic dmDys differentially expressed 1281 genes, whereas the hypoxic WT flies differentially expressed 56 genes. Interestingly, a number of genes (e.g. heat shock proteins) were discordantly regulated in response to CH between dmDys and WT. We tested the possibility that the disparate molecular responses of dystrophin-deficient tissues to CH could adversely affect muscle by performing functional assays in vivo. Normoxic and CH WT and dmDys flies were challenged with acute hypoxia and time-to-recover determined as well as subjected to climbing tests. Impaired performance was noted for CH-dmDys compared to normoxic dmDys or WT flies (rank order: Normoxic-WT ≈ CH-WT> Normoxic-dmDys> CH-dmDys). These data suggest that dystrophin-deficiency is associated with a disparate, pathological hypoxic stress response(s) and is more sensitive to hypoxia induced muscle dysfunction in vivo. We hypothesize that targeting/correcting the disparate molecular response(s) to hypoxia may offer a novel therapeutic strategy in DMD.
杜氏肌营养不良症(DMD)是一种严重的进行性肌病,由 DMD 基因突变导致抗肌萎缩蛋白缺失引起。由于呼吸肌不断坏死,晚期 DMD 与呼吸功能不全和慢性缺氧(CH)有关。为了了解 CH 对体内缺乏 dystrophin 的肌肉的影响,我们在将果蝇模型(dmDys)送往麦金利山(Denali/McKinley,海拔 6194 米)山顶的 16 天上升过程中使其暴露于 CH 之下。此外,dmDys 和野生型(WT)果蝇也在高海拔缺氧的实验室模拟中暴露于 CH 之下。使用 Affymetrix GeneChips®进行表达谱分析,并使用 qPCR 进行验证。低氧 dmDys 差异表达了 1281 个基因,而低氧 WT 果蝇差异表达了 56 个基因。有趣的是,许多基因(例如热休克蛋白)在 dmDys 和 WT 对 CH 的反应中呈现出不同的调节。我们通过在体内进行功能测定来测试缺乏 dystrophin 的组织对 CH 的不同分子反应可能对肌肉产生不利影响的可能性。对正常氧和 CH 的 WT 和 dmDys 果蝇进行急性低氧挑战,并确定恢复时间以及进行攀爬测试。与正常氧 dmDys 或 WT 果蝇相比,CH-dmDys 的表现受损(等级顺序:正常氧-WT ≈ CH-WT>正常氧-dmDys> CH-dmDys)。这些数据表明,缺乏 dystrophin 与不同的、病理性的低氧应激反应有关,并且对体内缺氧诱导的肌肉功能障碍更为敏感。我们假设针对/纠正对低氧的不同分子反应可能为 DMD 提供一种新的治疗策略。