The Potential Role of Cycloastragenol in Promoting Diabetic Wound Repair In Vitro.

Yi Cao,Fengling Xing,Qiongxiang Ge,Xiaohong Yang,Li Xu,Yuan Dong,Hongbin Luo

doi:10.1155/2019/7023950

Abstract

Background Refractory wound healing is a severe complication of diabetes with a significant socioeconomic burden. Whereas current therapies are insufficient to accelerate repair, stem cell-based therapy is increasingly recognized as an alternative that improves healing outcomes. The aim of the present study is to explore the role of cycloastragenol (CAG), a naturally occurring compound in Astragali Radix, in ameliorating refractory cutaneous wound healing in vitro, which may provide a new insight into therapeutic strategy for diabetic wounds. Methods Human epidermal stem cells (EpSCs) obtained from nine patients were exposed to CAG, with or without DKK1 (a Wnt signaling inhibitor). A lentiviral short hairpin RNA (shRNA) system was used to establish the telomerase reverse transcriptase (TERT) and β-catenin knockdown cell line. Cell counting kit-8, scratch wound healing, and transwell migration assay were used to determine the effects of CAG in cell growth and migration. The activation of TERT, β-catenin, and c-Myc was determined using real-time qPCR and western blot analysis. Chromatin immunoprecipitation (ChIP) was performed to evaluate the associations among CAG, TERT, and Wnt/β-catenin signals. Results CAG not only promoted the proliferation and migration ability of EpSCs but also increased the expression levels of TERT, β-catenin, c-Myc. These effects of CAG were most pronounced at a dose of 0.3 μM. Notably, the CAG-promoted proliferative and migratory abilities of EpSCs were abrogated in TERT and β-catenin-silenced cells. In addition, the ChIP results strongly suggested that CAG-modulated TERT was closely associated with the activation of Wnt/β-catenin signaling. Conclusion Our data indicate that CAG is a TERT activator of EpSCs and is associated with their proliferation and migration, a role it may play through the activation of Wnt/β-catenin signaling.

Highlights

In recent years, wound repair, especially for chronic nonhealing wounds, has emerged as a global public health issue that concerns physical and mental health in both type 1 and type 2 diabetes [1]
Cell migratory ability was observed using both transwell migration assay (Figure 2(c)) and scratch wound healing assay (Figure 2(d)). ese observations showed that CAG effectively promoted the migratory ability of epidermal stem cells (EpSCs) at a dose of 0.3 μM up to 24 h. ese data strongly support the hypothesis that CAG can play an important role in promoting the proliferation and migration of human primary EpSCs
In the in vitro experiment, we found that the expression of telomerase reverse transcriptase (TERT) was considerably raised in EpSCs subjected to 0.3 μM of CAG EpSCs (Figure 3(a))

Summary

Introduction

Wound repair, especially for chronic nonhealing wounds, has emerged as a global public health issue that concerns physical and mental health in both type 1 and type 2 diabetes [1]. BioMed Research International tissue, is increasingly recognized as a crucial component in diabetic wound healing. Stem cell-based therapy is gaining recognition as a way to improve healing outcomes [6]. Whereas current therapies are insufficient to accelerate repair, stem cell-based therapy is increasingly recognized as an alternative that improves healing outcomes. E aim of the present study is to explore the role of cycloastragenol (CAG), a naturally occurring compound in Astragali Radix, in ameliorating refractory cutaneous wound healing in vitro, which may provide a new insight into therapeutic strategy for diabetic wounds. Human epidermal stem cells (EpSCs) obtained from nine patients were exposed to CAG, with or without DKK1 (a Wnt signaling inhibitor). CAG promoted the proliferation and migration ability of EpSCs and increased the expression levels of TERT, β-catenin, c-Myc

Methods

Results

Conclusion