Sulforaphane increases Nrf2 expression and protects alveolar epithelial cells against injury caused by cigarette smoke extract

Jiao,Zongxian; Chang,Jiachen; Li,Jing; Nie,Dengmei; Cui,Huijuan; Guo,Dongfang

doi:10.3892/mmr.2017.6700

Sulforaphane increases Nrf2 expression and protects alveolar epithelial cells against injury caused by cigarette smoke extract

Authors:
- Zongxian Jiao
- Jiachen Chang
- Jing Li
- Dengmei Nie
- Huijuan Cui
- Dongfang Guo
View Affiliations

Affiliations: Research Institute of Pathology, School of Basic Medical Sciences, Lanzhou University, Lanzhou, Gansu 730000, P.R. China
Published online on: June 6, 2017 https://doi.org/10.3892/mmr.2017.6700
Pages: 1241-1247
Copyright: © Jiao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Cigarette smoking is a primary risk factor for chronic obstructive pulmonary disease (COPD), as it damages epithelial cells through a variety of mechanisms. Sulforaphane (SFN) is an antioxidant agent, which exerts protective effects against cell damage by activating the nuclear factor erythroid 2 like 2 (NFE2L2; Nrf2). The present study was undertaken to investigate the effects and underlying mechanisms of SFN in preventing cigarette smoke extract (CSE)‑induced oxidative damage to RLE‑6TN rat lung epithelial cells. MTT assay was used to determine the cytotoxicity of SFN and CSE. The effect of SFN and CSE on cell cycle progression, apoptosis and intracellular reactive oxygen species (ROS) levels were analyzed using flow cytometry. Reverse transcription‑quantitative polymerase chain reaction and western blotting were used to quantify mRNA and protein expression levels of Nrf2 respectively. SFN protected RLE‑6TN cells from oxidative damage, potentially via increasing Nrf2 expression and reducing ROS levels. In addition, SFN attenuated G1 phase cell cycle arrest and abrogated apoptosis. Therefore, SFN protected alveolar epithelial cells against CSE‑induced oxidative injury by upregulating Nrf2 expression. The results of the present study may provide theoretical support for the clinical use of SFN in patients with COPD.

View Figures

View References

1	Jansson SA, Backman H, Stenling A, Lindberg A, Rönmark E and Lundbäck B: Health economic costs of COPD in Sweden by disease severity-has it changed during a ten years period? Respir Med. 107:1931–1938. 2013. View Article : Google Scholar : PubMed/NCBI
2	Decramer M, Janssens W and Miravitlles M: Chronic obstructive pulmonary disease. Lancet. 379:1341–1351. 2012. View Article : Google Scholar : PubMed/NCBI
3	Tamimi A, Serdarevic D and Hanania NA: The effects of cigarette smoke on airway inflammation in asthma and COPD: Therapeutic implications. Respir Med. 106:319–328. 2012. View Article : Google Scholar : PubMed/NCBI
4	Zhang H, Shih A, Rinna A and Forman HJ: Exacerbation of tobacco smoke mediated apoptosis by resveratrol: An unexpected consequence of its antioxidant action. Int J Biochem Cell Biol. 43:1059–1064. 2011. View Article : Google Scholar : PubMed/NCBI
5	Goven D, Boutten A, Leçon-Malas V, Boczkowski J and Bonay M: Prolonged cigarette smoke exposure decreases heme oxygenase-1 and alters Nrf2 and Bach1 expression in human macrophages: Roles of the MAP kinases ERK(1/2) and JNK. FEBS Lett. 583:3508–3518. 2009. View Article : Google Scholar : PubMed/NCBI
6	Regoli F and Giuliani ME: Oxidative pathways of chemical toxicity and oxidative stress biomarkers in marine organisms. Mar Environ Res. 93:106–117. 2014. View Article : Google Scholar : PubMed/NCBI
7	Tasaki M, Kuroiwa Y, Inoue T, Hibi D, Matsushita K, Kijima A, Maruyama S, Nishikawa A and Umemura T: Lack of nrf2 results in progression of proliferative lesions to neoplasms induced by long-term exposure to non-genotoxic hepatocarcinogens involving oxidative stress. Exp Toxicol Pathol. 66:19–26. 2014. View Article : Google Scholar : PubMed/NCBI
8	Cano M, Thimmalappula R, Fujihara M, Nagai N, Sporn M, Wang AL, Neufeld AH, Biswal S and Handa JT: Cigarette smoking, oxidative stress, the anti-oxidant response through Nrf2 signaling and Age-related macular degeneration. Vision Res. 50:652–664. 2010. View Article : Google Scholar : PubMed/NCBI
9	Rangasamy T, Cho CY, Thimmulappa RK, Zhen L, Srisuma SS, Kensler TW, Yamamoto M, Petrache I, Tuder RM and Biswal S: Genetic ablation of Nrf2 enhances susceptibility to cigarette smoke-induced emphysema in mice. J Clin Invest. 114:1248–1259. 2004. View Article : Google Scholar : PubMed/NCBI
10	Suzuki M, Betsuyaku T, Ito Y, Nagai K, Nasuhara Y, Kaga K, Kondo S and Nishimura M: Down-regulated NF-E2-related factor 2 in pulmonary macrophages of aged smokers and patients with chronic obstructive pulmonary disease. Am J Respir Cell Mol Biol. 39:673–682. 2008. View Article : Google Scholar : PubMed/NCBI
11	Malhotra D, Thimmulappa R, Navas-Acien A, Sandford A, Elliott M, Singh A, Chen L, Zhuang X, Hogg J, Pare P, et al: Expression of concern: Decline in NRF2-regulated antioxidants in chronic obstructive pulmonary disease lungs due to loss of its positive regulator, DJ-1. Am J Respir Crit Care Med. 178:592–604. 2008. View Article : Google Scholar : PubMed/NCBI
12	Ping Z, Liu W, Kang Z, Cai J, Wang Q, Cheng N, Wang S, Wang S, Zhang JH and Sun X: Sulforaphane protects brains against hypoxic-ischemic injury through induction of Nrf2-dependent phase 2 enzyme. Brain Res. 1343:178–185. 2010. View Article : Google Scholar : PubMed/NCBI
13	Guerrero-Beltrán CE, Calderón-Oliver M, Pedraza-Chaverri J and Chirino YI: Protective effect of sulforaphane against oxidative stress: Recent advances. Exp Toxicol Pathol. 64:503–508. 2012. View Article : Google Scholar : PubMed/NCBI
14	Bergström P, Andersson HC, Gao Y, Karlsson JO, Nodin C, Anderson MF, Nilsson M and Hammarsten O: Repeated transient sulforaphane stimulation in astrocytes leads to prolonged Nrf2-mediated gene expression and protection from superoxide-induced damage. Neuropharmacology. 60:343–353. 2011. View Article : Google Scholar : PubMed/NCBI
15	Hu R, Hebbar V, Kim BR, Chen C, Winnik B, Buckley B, Soteropoulos P, Tolias P, Hart RP and Kong AN: In vivo pharmacokinetics and regulation of gene expression profiles by isothiocyanate sulforaphane in the rat. J Pharmacol Exp Ther. 310:263–271. 2004. View Article : Google Scholar : PubMed/NCBI
16	Kobayashi A, Kang MI, Okawa H, Ohtsuji M, Zenke Y, Chiba T, Igarashi K and Yamamoto M: Oxidative stress sensor Keap1 functions as an adaptor for Cul3-based E3 ligase to regulate proteasomal degradation of Nrf2. Mol Cell Biol. 24:7130–7139. 2004. View Article : Google Scholar : PubMed/NCBI
17	Iida K, Itoh K, Kumagai Y, Oyasu R, Hattori K, Kawai K, Shimazui T, Akaza H and Yamamoto M: Nrf2 is essential for the chemopreventive efficacy of oltipraz against urinary bladder carcinogenesis. Cancer Res. 64:6424–6431. 2004. View Article : Google Scholar : PubMed/NCBI
18	Xu C, Huang MT, Shen G, Yuan X, Lin W, Khor TO, Conney AH and Kong AN: Inhibition of 7,12-dimethylbenz(a)anthracene-induced skin tumorigenesis in C57BL/6 mice by sulforaphane is mediated by nuclear factor E2-related factor 2. Cancer Res. 66:8293–8296. 2006. View Article : Google Scholar : PubMed/NCBI
19	Burkhardt BR, Lyle R, Qian K, Arnold AS, Cheng H, Atkinson MA and Zhang YC: Efficient delivery of siRNA into cytokine-stimulated insulinoma cells silences Fas expression and inhibits Fas-mediated apoptosis. FEBS Lett. 580:553–560. 2006. View Article : Google Scholar : PubMed/NCBI
20	Oda K, Yumoto R, Nagai J, Katayama H and Takano M: Mechanism underlying insulin uptake in alveolar epithelial cell line RLE-6TN. Eur J Pharmacol. 672:62–69. 2011. View Article : Google Scholar : PubMed/NCBI
21	Nadigel J, Audusseau S, Baglole CJ, Eidelman DH and Hamid Q: IL-8 production in response to cigarette smoke is decreased in epithelial cells from COPD patients. Pulm Pharmacol Ther. 26:596–602. 2013. View Article : Google Scholar : PubMed/NCBI
22	Thorne D and Adamson J: A review of in vitro cigarette smoke exposure systems. Exp Toxicol Pathol. 65:1183–1193. 2013. View Article : Google Scholar : PubMed/NCBI
23	Bryan HK, Olayanju A, Goldring CE and Park BK: The Nrf2 cell defence pathway: Keap1-dependent and -independent mechanisms of regulation. Biochem Pharmacol. 85:705–717. 2013. View Article : Google Scholar : PubMed/NCBI
24	Kosmider B, Messier EM, Chu HW and Mason RJ: Human alveolar epithelial cell injury induced by cigarette smoke. PLoS One. 6:e260592011. View Article : Google Scholar : PubMed/NCBI