Gene Reports 26 (2022) 101522 Available online 22 January 2022 2452-0144/© 2022 Elsevier Inc. All rights reserved. N-Acetyl-L-cysteine effects on oxidative stress-induced high glucose-cultured human dermal fbroblasts Mohammad Hossein Rashnavadi , Zahra Tahmasebi , Zeinab Piravar * , Azadeh Mohammad Gholi Department of Biology, Faculty of Sciences, Central Tehran Branch, Islamic Azad University, Tehran, Iran A R T I C L E INFO Edited by Hee-Jeong Im Sampen Keywords: High glucose Oxidative stress Nrf2 Catalase Glutathione N-acetyl cysteine ABSTRACT Background: Wound healing in diabetic patients is delayed, but the mechanism of this impairment is unclear. We sought to investigate molecular changes in human dermal fbroblasts (HDFs) in a high-glucose state and improving effect of N-acetyl-L-cysteine (NAC). Methods: HDFs were cultured in 5.5, 25, 50, and 75 mM glucose concentrations for 72 h. Cell proliferation and migration were examined via in vitro scratch and 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assays. Cytotoxicity was determined through the measurement of lactate dehydrogenase (LDH). The level of intracellular reactive oxygen species (ROS) was also measured in the HDFs. Oxidative stress markers of nuclear factor erythroid 2 (NFE2)- related factor 2 (NRF2), Catalase (CAT) and Glutathione (GSH) were quantifed with Real time polymerase chain reaction (PCR). The antioxidant effect of NAC, 1 mM was examined to evaluate the involvement of ROS and LDH and oxidative markers in the glucose effects on the HDFs. Results: The in vitro scratch assay showed that high glucose concentrations signifcantly reduced fbroblast migration and proliferation at 12, 24, and 48 h. The MTT assay revealed a decline in cell viability in 50 and 75 mM glucose concentrations. A signifcant increase in LDH and ROS levels was observed in the HDFs incubated in 50 and 75 mM glucose concentrations after 72 h. mRNA level of Nrf2, CAT and GSH were downregulated. The antioxidant addition of NAC reduced the inhibitory effect of the high-glucose state on the migration and pro- liferation of the HDFs. Conclusions: A high-glucose state impairs the in vitro proliferation and migration of HDFs and may, therefore, induce increased oxidative stress and cellular dysfunction. The antioxidant effect of NAC ameliorates the damaging impact of a high-glucose state. 1. Introduction Diabetes is a disease characterized by elevated blood sugar levels. The impaired metabolism of glucose, lipids, and proteins in diabetes increases the risk of affiction to other diseases such as retinopathy, neuropathy, nephropathy, and cardiovascular disease (Pang et al., 2016). Delayed wound healing is another complication of diabetes, which may cause chronic ulcers and even amputation (Liu et al., 2011), with the latter not only diminishing the quality of life of the victims but also imposing substantial economic burdens on communities. Wound healing is a complex process that involves various chemical and bio- logical interactions between cells, growth and infammatory factors, and extracellular matrix compounds (Kulprachakarn et al., 2017). Hyper- glycemia delays wound healing by creating an infammatory state and inhibiting tissue angiogenesis. Laboratory studies have indicated that hyperglycemia stymies the migration, proliferation, and synthesis of vital factors such as collagen in cells and amplifes the rate of apoptosis (Chen et al., 2018). Wound healing requires coordination between Abbreviations: HDFs, Human Dermal Fibroblasts; NAC, N-Acetyl-L-cysteine; MTT, 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide; LDH, lactate dehydrogenase; ROS, reactive oxygen species; PCR, polymerase chain reaction; CAT, Catalase; NRF2, nuclear factor erythroid 2 (NFE2)- related factor 2; DMEM, Dulbecco's Modifed Eagle's Medium; FBS, fetal bovine serum; AUC, area under the percent migration curve; OD, optical density; DMSO, dimethyl sulfoxide; DCFDA, dichlorodihydrofuorescein diacetate. * Corresponding author at: Department of Biology, Faculty of Sciences, Central Tehran Branch, Islamic Azad University, Ashraf Isfahani Highway, Imam Hassan Blvd., Orchid code: 0000-0001-8949-362X, Tehran 1955847881, Iran. E-mail address: saba.piravar@gmail.com (Z. Piravar). Contents lists available at ScienceDirect Gene Reports journal homepage: www.elsevier.com/locate/genrep https://doi.org/10.1016/j.genrep.2022.101522 Received 2 October 2021; Received in revised form 23 December 2021; Accepted 14 January 2022 Downloaded from https://iranpaper.ir https://www.tarjomano.com https://www.tarjomano.com