Glycitein alleviates inflammation and apoptosis in keratinocytes via ROS-associated PI3K-Akt signalling pathway.

Abstract

A chronic inflammatory skin disorder, psoriasis, affects 2-3% of people worldwide. A bioactive substance, glycitein (GCN), has several pharmacological characteristics. This work aims to evaluate the effects of GCN on the in vitro proliferation and death of human HaCaT keratinocytes. An in vitro model was created to simulate psoriatic features utilizing HaCaT keratinocytes activated by M5 cytokines. The 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide test was used to quantify cell viability, whereas the BrdU assay was used to assess the proliferation rate. Using a DCFH-DA probe and an Annexin V-FITC/propidium iodide detection kit, flow cytometry was used to examine the generation of reactive oxygen species (ROS) and apoptosis, respectively. Western blot and quantitative polymerase chain reaction were employed to determine the amounts of phosphorylated Akt (p-Akt) and Akt proteins. GCN dramatically decreased the inflammation and hyperproliferation that cytokines caused in HaCaT keratinocytes. The alteration of mitochondrial membrane potential promoted apoptosis and caused cell cycle arrest at the sub-G1 phase, which indicates apoptotic DNA fragmentation. The suppression of the PI3K/Akt signalling pathway was linked to increased intracellular ROS levels brought on by GCN therapy. These results imply that GCN reduces inflammation and keratinocyte hyperproliferation by controlling cell cycle progression and apoptosis via ROS-associated inhibition of the PI3K/Akt pathway.