Anthocyanins Extracted from <i>Aronia melanocarpa</i> Protect SH-SY5Y Cells against Amyloid-beta (1–42)-Induced Apoptosis by Regulating Ca²⁺ Homeostasis and Inhibiting Mitochondrial Dysfunction

Abstract

We investigated the cytoprotective effects of anthocyanins in Aronia melanocarpa against apoptosis induced by Aβ_1-42, a key mediator of AD pathophysiology. We measured intracellular calcium with a colorimetric kit, cellular apoptosis with DAPI, intracellular ROS with the fluorescent marker 2,3-dimethoxy-1,4-naphthoquinone, mitochondrial membrane potential with JC-1, and ATP with a colorimetric kit. Gene transcription and protein expression levels of calmodulin, cytochrome c, caspase-9, cleaved caspase-3, Bcl-2, and Bax were analyzed by RT-PCR and Western blotting. The results showed that pretreatment with anthocyanins significantly inhibited Aβ_1-42-induced apoptosis, decreased intracellular calcium and ROS, and increased ATP and mitochondrial membrane potential. RT-PCR and Western blotting revealed that anthocyanins upregulated the gene transcription and protein expression of calmodulin and Bcl-2 and downregulated those of cytochrome c, caspase-9, cleaved caspase-3, and Bax. A. melanocarpa anthocyanins protected SH-SY5Y cells against Aβ_1-42-induced apoptosis by regulating Ca²⁺ homeostasis and apoptosis-related genes and inhibiting mitochondrial dysfunction.