Increased proteolytic activity has been widely associated with skeletal muscle atrophy. However, elevated proteolysis is also critical for the maintenance of intracellular homeostasis. In this study, we aimed to investigate the significance of autophagy in obesity-induced muscle atrophy and clarify the mechanism involved. First, high-fat diet (HFD)-fed rats were administered vehicle or chloroquine (CQ), an autophagy inhibitor, and we found that HFD inhibited autophagic flux and reduced myofiber size and function in rats. Additionally, the expression levels of MyoD were decreased whereas those of Atrogin-1 were increased in rats fed a HFD. Sustained autophagy inhibition by CQ exacerbated HFD-induced muscular damage and changes in the expression of Atrogin-1 and MyoD. Similar effects were reproduced in vitro in myotubes, which exhibited increased levels of autophagy-related proteins, but the resultant autophagic flux was reduced following exposure to palmitic acid (PA)-conditioned medium. Moreover, PA significantly decreased MyoD levels and induced Atrogin-1 expression, leading to progressive myotube atrophy; this phenomenon was aggravated by CQ but alleviated by the autophagy activator rapamycin. Taken together, these in vivo and in vitro findings suggest that autophagic flux is blocked in skeletal muscle of individuals with high lipid, and autophagy mediates high lipid-induced muscle atrophy by affecting muscle degradation and regeneration. (C) 2020 Elsevier Inc. All rights reserved.