Inhibition of CTR1 expression improves hypoxia/reoxygenation-induced myoblast injury by blocking cuproptosis

• Published in: Biochemical and biophysical research communications 2024-11, Vol.735, p.150804, Article 150804
• Main Authors: Fu, Dong-ge, He, Jing-zi, Mu, Qi-chen, Huo, Yan-fang, Zhang, Ning-mei, Zhang, Le, Hua, Shu, Gao, Bao-quan
• Format: Article
• Language: English
• Subjects: Animals; Cation Transport Proteins - genetics; Cation Transport Proteins - metabolism; Cell Hypoxia - drug effects; Cell Line; Cell Survival - drug effects; Copper - metabolism; Copper Transporter 1 - metabolism; CTR1; Cuproptosis; Ischemia/reperfusion; Mice; Myoblasts - drug effects; Myoblasts - metabolism; Oxygen - metabolism; Reperfusion Injury - metabolism; Reperfusion Injury - pathology; Skeletal muscle
• ISSN: 0006-291X; 1090-2104; 1090-2104
• DOI: 10.1016/j.bbrc.2024.150804

Skeletal muscle ischemia-reperfusion injury (IRI) is a common severe disease with a complex pathological process. This study found that copper chloride (CuCl2) inhibited cell viability in a concentration dependent manner, increased intracellular copper levels and downregulated copper transporter 1 (CTR1) expression. CTR1 upregulation promoted copper uptake by myoblasts and then enhanced cuproptosis, leading to a significant increase in the levels of dihydrolipoamide S-acetyltransferase (DLAT) oligomers, while a significant decrease in the levels of lipoylated (Lip)-dihydrolipoamide S-succinyltransferase (DLST) and Lip-DLAT, ultimately inhibiting cell viability and inducing cell injury. Inducing cuproptosis with elesclomol plus CuCl2 (ES + Cu) further confirmed that “ES + Cu” treatment significantly reduced the contents of adenosine triphosphate (ATP) and glutathione (GSH), decreased the activities of mitochondrial complex I and III, and increased the contents of lactate (LA), malondialdehyde (MDA), creatine kinase (CK) and lactate dehydrogenase (LDH); when tetrathiomolybdate (TTM) was added to inhibit cuproptosis, myoblast injury was recovered significantly. Meanwhile, hypoxia/reoxygenation (H/R) induced CTR1 expression, increased the levels of intracellular copper, DLAT oligomers, LA, MDA, CK and LDH, reduced the levels of Lip-DLST, Lip-DLAT, ATP and GSH, and weakened the activities of mitochondrial complex I and III; after knocking down CTR1 expression, the levels of intracellular copper and the activation of cuproptosis pathway were decreased, and cell viability, injury and inflammation levels were significantly improved. Therefore, cuproptosis can promote myoblast injury, while H/R enhances copper uptake by inducing CTR1 expression, thereby enhancing cuproptosis and inducing cell injury, indicating that cuproptosis is a new mechanism of H/R-induced myoblast injury. •CTR1 overexpression combined with copper treatment inhibits skeletal muscle cell viability by promoting copper uptake.•CTR1 overexpression plus copper treatment induces skeletal muscle cell cuproptosis.•Inducing cuproptosis promotes skeletal muscle cell injury.•H/R treatment induces cuproptosis by upregulating the expression of CTR1.•H/R treatment enhances skeletal muscle cell injury and inflammation by inducing cuproptosis.