Effect of Prunus yedoensis Matsumura extract on the gene expression in HaCaT cells

Abstract

The bark of Prunus yedoensis (PY) is used in herbal remedies for the treatment of inflammatory skin disorders. Previous studies have reported the antioxidative, anti-inflammatory, anticancer, and vasorelaxant effects of PY; however, the safety and toxicity of PY extracts have not yet been reported. In the present study, we explored whether PY bark extract induced toxicity in HaCaT, a human keratinocyte cell line. HaCaT cells were treated with PYE, the ethyl acetate (EtOAc) fraction of the crude extract (produced from 80% EtOH) of PY, for 24 h and cytotoxicity and ROS generation were determined. In addition, we evaluated the effect of a high concentration of PYE on the transcriptional profile to determine the action mechanism of the toxicity to HaCaT cells. PYE treatment significantly induced cytotoxicity in HaCaT cells in a dose-dependent manner. At 313 μg/mL, PYE generated intracellular ROS and up-regulated immediate early genes and oxidative stress-related genes, indicating the activation of stress signals by PYE. Moreover, PYE significantly changed the expression of genes that promote ECM degradation, inflammatory responses, and keratinocyte differentiation. The present study demonstrated that PYE at 313 μg/mL, a concentration which is not cytotoxic but generates intracellular ROS, affected gene regulation in HaCaT human keratinocytes through multiple toxicity mechanisms and could therefore disturb skin homeostasis or lead to skin aging. We propose that for the safe use of PYE in herbal cosmetic or medicines, further evaluations of the toxicity and safety in the skin must be performed.