Altered trace element homeostasis is associated with diabetic complications, and studies have shown elevated copper levels in the serum of individuals with type 1 & 2 diabetes. Copper chelation has been shown to be beneficial by preventing or reversing diabetic organ damage and developing as a new treatment strategy for treating diabetic complications. Diabetic retinopathy is the major vision-threatening complication of diabetes. Recent studies have reported copper to be elevated in the serum of patients with diabetic retinopathy. Here in this study, we attempt to unravel the role of copper chelator penicillamine in retinal pigment epithelial cells exposed to high glucose (HG) and copper as a model for diabetic retinopathy. We have found that high glucose by itself and along with copper alters the mitochondrial morphology, reduces the expression of the mitochondrial fusion protein 2 (MFN2), and induces endoplasmic reticulum (ER) stress and inflammation. Copper chelation with penicillamine reduced all these changes in mitochondria, thereby rescuing the cells from mitochondrial damage and inflammation.Keywords:Copper transporter 1; High glucose; Mitofusion 2; Retinal pigment epithelial cells.