Dillingham Blythe C, Benny Klimek Margaret E, Gernapudi Ramkishore, Rayavarapu Sree, Gallardo Eduard, Van der Meulen Jack H, Jordan Sarah, Ampong Beryl, Gordish-Dressman Heather, Spurney Christopher F, Nagaraju Kanneboyina
Research Center for Genetic Medicine, Children's National Medical Center, 111 Michigan Ave NW, Washington, D.C., USA.
Institut de Recerca Hospital de la Santa Creu i Sant Pau. U.A.B.C./Pare Claret, 167 08025 Barcelona, Spain.
J Neurol Sci. 2015 Sep 15;356(1-2):157-62. doi: 10.1016/j.jns.2015.06.042. Epub 2015 Jun 24.
The dysferlin-deficient A/J mouse strain represents a homologous model for limb-girdle muscular dystrophy 2B. We evaluated the disease phenotype in 10 month old A/J mice compared to two dysferlin-sufficient, C57BL/6 and A/JOlaHsd, mouse lines to determine which functional end-points are sufficiently sensitive to define the disease phenotype for use in preclinical studies in the A/J strain. A/J mice had significantly lower open field behavioral activity (horizontal activity, total distance, movement time and vertical activity) when compared to C57BL/6 and A/JoIaHsd mice. Both A/J and A/JOIaHsd mice showed decreases in latency to fall with rotarod compared to C57BL/6. No changes were detected in grip strength, force measurements or motor coordination between these three groups. Furthermore, we have found that A/J muscle shows significantly increased levels of the pro-inflammatory cytokine TNF-α when compared to C57BL/6 mice, indicating an activation of NF-κB signaling as part of the inflammatory response in dysferlin-deficient muscle. Therefore, we assessed the effect of celastrol (a potent NF-κB inhibitor) on the disease phenotype in female A/J mice. Celastrol treatment for four months significantly reduced the inflammation in A/J muscle; however, it had no beneficial effect in improving muscle function, as assessed by grip strength, open field activity, and in vitro force contraction. In fact, celastrol treated mice showed a decrease in body mass, hindlimb grip strength and maximal EDL force. These findings suggest that inhibition of inflammation alone may not be sufficient to improve the muscle disease phenotype in dysferlin-deficient mice and may require combination therapies that target membrane stability to achieve a functional improvement in skeletal muscle.
dysferlin基因缺陷的A/J小鼠品系是肢带型肌营养不良2B的同源模型。我们评估了10月龄A/J小鼠与两种dysferlin基因正常的C57BL/6和A/JOlaHsd小鼠品系的疾病表型,以确定哪些功能终点对定义A/J品系临床前研究中的疾病表型具有足够的敏感性。与C57BL/6和A/JoIaHsd小鼠相比,A/J小鼠的旷场行为活动(水平活动、总距离、移动时间和垂直活动)显著降低。与C57BL/6相比,A/J和A/JOIaHsd小鼠在转棒试验中的跌落潜伏期均缩短。这三组之间在握力、力量测量或运动协调性方面未检测到变化。此外,我们发现与C57BL/6小鼠相比,A/J小鼠肌肉中促炎细胞因子TNF-α水平显著升高,表明NF-κB信号通路被激活,这是dysferlin基因缺陷肌肉炎症反应的一部分。因此,我们评估了雷公藤红素(一种有效的NF-κB抑制剂)对雌性A/J小鼠疾病表型的影响。雷公藤红素治疗四个月可显著减轻A/J小鼠肌肉的炎症;然而,通过握力、旷场活动和体外力量收缩评估,它对改善肌肉功能没有有益作用。事实上,雷公藤红素治疗的小鼠体重、后肢握力和最大EDL力量均下降。这些发现表明,仅抑制炎症可能不足以改善dysferlin基因缺陷小鼠的肌肉疾病表型,可能需要针对膜稳定性的联合治疗来实现骨骼肌功能的改善。