Zhang Yue-Yue, Yang Man, Bao Jin-Fang, Gu Li-Jie, Yu Hong-Lei, Yuan Wei-Jie
Department of Nephrology, Shanghai General Hospital, Shanghai Jiaotong University School of Medcine, Haining Road 100rd, Hongkou Distrinct, Shanghai, 200080, China.
BMC Nephrol. 2018 Feb 27;19(1):45. doi: 10.1186/s12882-018-0836-2.
Accelerated muscle atrophy is associated with a three-fold increase in mortality in chronic kidney disease (CKD) patients. It is suggested that hyperphosphatemia might contribute to muscle wasting, but the underlying mechanisms remain unclear. Although evidence indicates that autophagy is involved in the maintenance of muscle homeostasis, it is not known if high phosphate levels can result in activation of autophagy, leading to muscle protein loss.
Immortalized rat L6 myotubes were exposed to a high concentration of phosphate, with or without autophagy inhibition. Myotube atrophy was examined by phase contrast microscopy. Autophagic activity was assessed by measuring the expression of microtubule-associated protein 1 light chain 3 (LC3) and p62 using quantitative real-time polymerase chain reaction and western blot.
Phosphate induced cell atrophy in L6 myotubes in a dose- and time-dependent manner, and these responses were not associated with calcification or osteogenesis. Phosphate also dose- and time-dependently increased the LC3-II/LC3-I ratio. Inhibition of autophagy with wortmannin or knockdown of Atg5 significantly suppressed myotube atrophy caused by high phosphate concentration.
High phosphate concentration induces muscle cell atrophy through the activation of autophagy. Targeting autophagy could be a therapeutic strategy for preventing muscle wasting caused by hyperphosphatemia.
慢性肾脏病(CKD)患者中,加速的肌肉萎缩与死亡率增加三倍相关。有研究表明,高磷血症可能导致肌肉消耗,但潜在机制仍不清楚。尽管有证据表明自噬参与维持肌肉稳态,但尚不清楚高磷水平是否会导致自噬激活,进而导致肌肉蛋白流失。
将永生化大鼠L6肌管暴露于高浓度磷酸盐中,同时或不进行自噬抑制。通过相差显微镜检查肌管萎缩情况。使用定量实时聚合酶链反应和蛋白质免疫印迹法,通过测量微管相关蛋白1轻链3(LC3)和p62的表达来评估自噬活性。
磷酸盐以剂量和时间依赖性方式诱导L6肌管细胞萎缩,且这些反应与钙化或骨生成无关。磷酸盐还以剂量和时间依赖性方式增加LC3-II/LC3-I比率。用渥曼青霉素抑制自噬或敲低Atg5可显著抑制高磷酸盐浓度引起的肌管萎缩。
高磷酸盐浓度通过激活自噬诱导肌肉细胞萎缩。针对自噬可能是预防高磷血症引起的肌肉消耗的一种治疗策略。