Histatin-1 alleviates high-glucose injury to skin keratinocytes through MAPK signaling pathway.

Abstract

Damage to keratinocytes and other skin cells in a high-glucose environment has been proven to be an important reason for the poor wound healing ability of chronic diabetes mellitus. Histatin-1 has been preliminarily proven to stimulate the wound healing process of the oral and non-oral mucosa and has been found to be related to the activation of extracellular signal-regulated kinase (ERK). The purpose of this study was to investigate the effect of histatin-1 on high-glucose-injured keratinocytes and the role of the Ras-Raf-MEK-ERK signaling pathway on the effect of histatin-1 to improve diabetic wound healing. A human keratinocyte model damaged by high glucose was constructed, cell proliferation was detected by the Cell Counting Kit-8 assay, and cell apoptosis was detected by flow cytometry. The expression level of 8-hydroxy-2'-deoxyguanosine (8-OHdG) was detected by ELISA, and the mitogen-activated protein kinase (MAPK) signaling pathway protein expression level was detected by Western blot. C-fos mRNA expression was detected by real-time PCR. The results indicated that histatin-1 promoted proliferation and reduced the rate of apoptosis and 8-OHdG content in keratinocytes with high-glucose injury. In addition, histatin-1 down-regulated MEK phosphorylation in keratinocytes with high-glucose injury. However, with the extension of the intervention, the effect of histatin-1 on c-fos mRNA expression was different. At the early stage of high-glucose injury (12 h), the expression of c-fos mRNA was not increased in high-glucose-injured keratinocytes treated with histatin-1 but then c-fos mRNA expression was gradually upregulated. Histatin-1 could alleviate keratinocyte injury caused by high glucose levels and promoted wound healing in vitro. In addition, histatin-1 could exert anti-apoptotic and antioxidant damage effects under high-glucose injury states. These effects of histatin-1 may be related to its regulation of the MAPK signaling pathway. Therefore, these findings provide an essential theoretical basis for histatin-1 to become a safe and effective new peptide biological agent to promote wound healing in patients with diabetes.