Gastrodin Alleviates Oxidative Stress-Induced Apoptosis and Cellular Dysfunction in Human Umbilical Vein Endothelial Cells via the Nuclear Factor-Erythroid 2-Related Factor 2/Heme Oxygenase-1 Pathway and Accelerates Wound Healing In Vivo.

Jialiang Lin,Jiansen Miao,Chen Liu,Xiangyang Wang,Weimin Zeng,Haiming Jin,Hao Lin,Fangzhou Qi,Yuhao Wu,Jianwei Wu,Yifeng Shi

doi:10.3389/fphar.2019.01273

Abstract

Aims: To explore the effect and mechanism of gastrodin (GAS) on human umbilical vein endothelial cells (HUVECs) apoptosis induced by oxidative stress and its function in wound healing. Main methods: HUVECs were incubated with tert-butyl hydroperoxide (TBHP) to induce endothelial cell dysfunction and GAS was used as a protector. Cell viability was detected by Counting Kit-8 (CCK-8). HUVECs apoptosis was evaluated by TUNEL assay and western blotting for cleaved caspase3 (C-caspase3) and other apoptosis-related proteins. Transwell migration assay, tube formation assay, and cell-matrix adhesion assay were performed to evaluated cell function of HUVECs. Transfection with nuclear factor-erythroid 2-related factor 2 (Nrf2) small interfering ribonucleic acid and western blotting for Nrf2, HO-1, and apoptosis-related proteins were performed to prove that Nrf2/HO-1 pathway is involved in the protective effects of GAS. The skin wound model of rat was used to assess the protective effects of GAS in vivo. Key Findings: The results show that treating HUVECs with GAS attenuated TBHP-induced apoptosis and cellular dysfunction, including cellular tube formation, migration, and adhesion. Mechanistically, we found that GAS protects HUVECs from TBHP-induced cellular apoptosis by activating the nuclear factor (erythroid-derived 2)-like 2 (Nrf2)/heme oxygenase 1 (HO-1) pathway. An in vivo study illustrated that the oral administration of GAS enhances vascularization in regenerated tissue and facilitates wound healing. Significance: The findings of this study demonstrated that GAS may serve as a potential agent that accelerates wound healing.

Highlights

The skin is a barrier to the outside world that protects the body from exogenous damage, and compared to other tissues, it is considered to be the most frequently injured tissue (Lee et al, 2006; Boer et al, 2016)
The results showed that the rate of TUNEL-positive cells and the level of C-caspase3, an apoptosis-related protein, were increased by tert-butyl hydroperoxide (TBHP) stimulation but reversed by GAS pretreatment in a dose-dependent manner (Figures 1D–G)
The results showed that TBHP significantly suppressed human umbilical vein endothelial cells (HUVECs) neovascularization, while GAS pretreatment protected against the effects of TBHP in a dose-dependent manner (Figures 3B, E)

Summary

Introduction

The skin is a barrier to the outside world that protects the body from exogenous damage, and compared to other tissues, it is considered to be the most frequently injured tissue (Lee et al, 2006; Boer et al, 2016). Cutaneous wound healing is a complex process that repairs and regenerates tissue structure and function; classically, this process can be divided into four highly integrated and overlapping phases: clot formation, inflammation, proliferation, and remodeling (Hosemann et al, 1991; Watelet et al, 2002; Mendonca and Coutinho-Netto, 2009; Velnar et al, 2009). This process is complex and involves the coordinated efforts of several cell types, including keratinocytes, fibroblasts, endothelial cells, macrophages, and platelets (Gurtner et al, 2008; Schultz et al, 2011; Wang et al, 2012). Protecting endothelial cells from oxidative stress-induced damage may be a promising therapeutic target for accelerating cutaneous wound healing

Methods

Results

Conclusion