AMPK Complex Activation Promotes Sarcolemmal Repair in Dysferlinopathy
AMPK Complex Activation Promotes Sarcolemmal Repair in Dysferlinopathy
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AMPK 复合物激活促进肌纤维蛋白病的肌膜修复
DOI：
10.1016/j.ymthe.2020.02.006
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发表时间：
2020
影响因子：
12.4
通讯作者：
Hayashi
中科院分区：
文献类型：
--
作者：
Ono Hiroya;Suzuki Naoki;Kanno Shin-ichiro;Kawahara Genri;Izumi Rumiko;Takahashi Toshiaki;Kitajima Yasuo;Osana Shion;Nakamura Naoko;Akiyama Tetsuya;Ikeda Kensuke;Shijo Tomomi;Mitsuzawa Shio;Nagatomi Ryoichi;Araki Nobukazu;Yasui Akira;Warita Hitoshi;Hayashi
indysferlin突变导致一组进行性、复发性遗传性肌营养不良，称为dysferlin病。利用重组蛋白和亲和纯化结合液相色谱-串联质谱（LC-MS/MS）的方法，我们发现AMP激活的蛋白激酶（AMPK）γ1与dysferlin的第三和第四C2结构域之间的区域结合。利用exvivolaser损伤实验，我们证明AMPK复合物在骨骼肌纤维肌膜损伤修复中是至关重要的。损伤诱导的AMPK复合物积累依赖于Ca 2+的存在，积累速率受dysferlin调节。此外，研究发现AMPKα的磷酸化对于质膜修复是必不可少的，并且用AMPK激活剂治疗挽救了在具有降低的dysferlin和dysferlin缺失小鼠纤维表达的永生化人肌管中观察到的膜修复损伤。最后，确定用AMPK激活剂二甲双胍治疗改善了dysferlin缺乏症的斑马鱼和小鼠模型的肌肉表型。这些发现表明AMPK复合物对于质膜修复是必需的，并且是dysferlin病的潜在治疗靶点。
Mutations indysferlinare responsible for a group of progressive, recessively inherited muscular dystrophies known as dysferlinopathies. Using recombinant proteins and affinity purification methods combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS), we found that AMP-activated protein kinase (AMPK)γ1 was bound to a region of dysferlin located between the third and fourth C2 domains. Usingex vivolaser injury experiments, we demonstrated that the AMPK complex was vital for the sarcolemmal damage repair of skeletal muscle fibers. Injury-induced AMPK complex accumulation was dependent on the presence of Ca2+, and the rate of accumulation was regulated by dysferlin. Furthermore, it was found that the phosphorylation of AMPKα was essential for plasma membrane repair, and treatment with an AMPK activator rescued the membrane-repair impairment observed in immortalized human myotubes with reduced expression of dysferlin and dysferlin-null mouse fibers. Finally, it was determined that treatment with the AMPK activator metformin improved the muscle phenotype in zebrafish and mouse models of dysferlin deficiency. These findings indicate that the AMPK complex is essential for plasma membrane repair and is a potential therapeutic target for dysferlinopathy.