The Regulation of GSTM5 in Oxidative Stress Induced by TNF-α in Human Bronchus Epithelial Cells_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

ZhaoYanxue ^1,2 , LiLi ¹ , YuanWeifeng ¹ , GuoZhenhui ³ ,  HuangWenjie ¹

1. Department of Respiratory Medicine, General Hospital of Guangzhou Military Command of PLA, Guangzhou, Guangdong, 510010, China;
2. ;
3. ;

Corresponding?author：

HuangWenjie, Email: huangyelu1029@vip.163.com

Keywords：

Glutathione S-transferase mu 5; Eukaryotic expression vector; Oxidative stress; Inflammation

DOI：

10.7507/1671-6205.2016066

Video：

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Objective To establish a cell inflammation model induced by tumor necrosis factor-α (TNF-α) in human bronchus epithelial cells, and investigate the effects of glutathione S-transferase mu 5 (GSTM5) on the inflammation and oxidative stress. Methods 16HBE cells were treated with TNF-α (10 ng/mL, 24 h) in the absence or presence of the constructed GSTM5 eukaryotic expression vector (1 μg/mL). The concentration of malondialdehyde (MDA) and total antioxidation capacity (T-AOC) were detected by colorimetric method. The survival rate of cells was assessed by the methyl thiazolyl tetrazolium (MTT) assay. The transcription level of NADPH oxidase-1 (NOX1), NOX2, NOX3, NOX4, NOX5, dual oxidase-1 (DUOX1) and DUOX2 were evaluated by RT-PCR. Western blot was performed to investigate the protein levels of NOX1 and NOX2. Results TNF-α simulation significantly increased the level of MDA in cells, and decreased the level of T-AOC and survival rate of 16HBE. When transfected with the GSTM5 eukaryotic expression vector, the concentration of MDA significantly decreased (P < 0.05), and the activation of T-AOC increased dramatically (P < 0.05). Consequently, the survival rate of 16HBE in the GSTM5 group improved (P < 0.05). The 16HBE cells transfected with the constructed GSTM5 eukaryotic expression vector had a lower transcription and protein levels of NOX1 and NOX2 (all P < 0.01). There were no significant changes in the mRNA expressions of NOX3, NOX4, NOX5, DUOX1 or DUOX2. Conclusion GSTM5 may down-regulate the transcription level of NOX1 and NOX2 to reduce the inflammation and oxidative stress induced by TNF-α.

Citation： Zhao Yanxue, Li Li, Yuan Weifeng, Guo Zhenhui, Huang Wenjie. The Regulation of GSTM5 in Oxidative Stress Induced by TNF-α in Human Bronchus Epithelial Cells. Chinese Journal of Respiratory and Critical Care Medicine, 2016, 15(3): 284-288. doi: 10.7507/1671-6205.2016066 Copy

1.	Prabhakaran P, Usatyuk PV, Gorshkova IA, et al. Acute respiratory distress syndrome. Indian Pediatr, 2010, 47:861-868.
2.	梁曉華, 黃勛, 田豐, 等.單免疫球蛋白白細胞介素1受體相關蛋白在脂多糖誘導的急性肺損傷小鼠肺組織中表達的研究.中國呼吸與危重監護雜志, 2013, 12:591-596.
3.	Chen H, Bai C, Wang X. The value of the lipopolysaccharide-induced acute lung injury model in respiratory medicine. Expert Rev Respir Med, 2010;4:773-783.
4.	Xu F, Fok TF, Yin J.Hyperoxia-induced lung injury in premature rat:description of a suitable model for the study of lung diseases in newborns. Chin Med J(Engl), 1998, 111:619-624.
5.	Chen S, Zheng S, Liu Z, et al. Endogeous sulfur dioxide protects against oleic acid-induced acute lung injury in association with inhibition of oxidative stress in rats. Lab Invest, 2015, 95:142-156.
6.	Corda S, Laplace C, Vicaut E, et al. Rapid reactive oxygen species production by mitochondria in endothelial cells exposed to tumor necrosis factor-α is mediated by ceramide. Am J Respir Cell Mol Biol, 2001, 24:762-768.
7.	龐戰軍, 周玫, 陳瑗, 主編.自由基醫學研究方法.北京:人民衛生出版社, 2000, 201-217.
8.	Rahman I. Oxidative stress, transcription factors and chromatin remodelling in lung inflammation. Biochem Pharmacol, 2002, 64:935-942.
9.	Tasaka S, Amaya F, Hashimoto S, et al. Roles of oxidants and redox signaling in the pathogenesis of acute respiratory distress syndrome. Antioxid Redox Signal, 2008, 10:739-753.
10.	邱嫻, 胡水旺, 徐俊, 等.A549細胞炎癥氧化應激模型Nox1啟動子差異結合蛋白的篩選.南方醫科大學學報, 2013, 5:703-707.
11.	Pendyala S, Usatyuk PV, Gorshkova IA, et al. Regulation of NADPH oxidase in vascular endothelium:the role of phospholipases, protein kinases, and cytoskeletal proteins. Antioxid Redox Signal, 2009, 11:841-860.
12.	Chua LS. A review on plant-based rutin extraction methods and its pharmacological activities. J Ethnopharmacol, 2013, 150:805-817.
13.	吳煒景, 李理, 袁偉鋒, 等.NF-κB p65基因在TNF-α誘導的肺泡上皮細胞氧化損傷中的作用.中國呼吸與危重監護雜志, 2012, 11:46-51.
14.	Maniatis NA, Orfanos SE. The endothelium in acute lung injury/acute respiratory distress syndrome. Curr Opin Crit Care, 2008, 14:22-30.
15.	Zhang WJ, Wei H, Tien YT, et al. Genetic ablation of phagocytic NADPH oxidase in mice limits TNFα-induced inflammation in the lungs but not other tissues. Free Radic Biol Med, 2011, 50:1517-1525.

1. Prabhakaran P, Usatyuk PV, Gorshkova IA, et al. Acute respiratory distress syndrome. Indian Pediatr, 2010, 47:861-868.
2. 梁曉華, 黃勛, 田豐, 等.單免疫球蛋白白細胞介素1受體相關蛋白在脂多糖誘導的急性肺損傷小鼠肺組織中表達的研究.中國呼吸與危重監護雜志, 2013, 12:591-596.
3. Chen H, Bai C, Wang X. The value of the lipopolysaccharide-induced acute lung injury model in respiratory medicine. Expert Rev Respir Med, 2010;4:773-783.
4. Xu F, Fok TF, Yin J.Hyperoxia-induced lung injury in premature rat:description of a suitable model for the study of lung diseases in newborns. Chin Med J(Engl), 1998, 111:619-624.
5. Chen S, Zheng S, Liu Z, et al. Endogeous sulfur dioxide protects against oleic acid-induced acute lung injury in association with inhibition of oxidative stress in rats. Lab Invest, 2015, 95:142-156.
6. Corda S, Laplace C, Vicaut E, et al. Rapid reactive oxygen species production by mitochondria in endothelial cells exposed to tumor necrosis factor-α is mediated by ceramide. Am J Respir Cell Mol Biol, 2001, 24:762-768.
7. 龐戰軍, 周玫, 陳瑗, 主編.自由基醫學研究方法.北京:人民衛生出版社, 2000, 201-217.
8. Rahman I. Oxidative stress, transcription factors and chromatin remodelling in lung inflammation. Biochem Pharmacol, 2002, 64:935-942.
9. Tasaka S, Amaya F, Hashimoto S, et al. Roles of oxidants and redox signaling in the pathogenesis of acute respiratory distress syndrome. Antioxid Redox Signal, 2008, 10:739-753.
10. 邱嫻, 胡水旺, 徐俊, 等.A549細胞炎癥氧化應激模型Nox1啟動子差異結合蛋白的篩選.南方醫科大學學報, 2013, 5:703-707.
11. Pendyala S, Usatyuk PV, Gorshkova IA, et al. Regulation of NADPH oxidase in vascular endothelium:the role of phospholipases, protein kinases, and cytoskeletal proteins. Antioxid Redox Signal, 2009, 11:841-860.
12. Chua LS. A review on plant-based rutin extraction methods and its pharmacological activities. J Ethnopharmacol, 2013, 150:805-817.
13. 吳煒景, 李理, 袁偉鋒, 等.NF-κB p65基因在TNF-α誘導的肺泡上皮細胞氧化損傷中的作用.中國呼吸與危重監護雜志, 2012, 11:46-51.
14. Maniatis NA, Orfanos SE. The endothelium in acute lung injury/acute respiratory distress syndrome. Curr Opin Crit Care, 2008, 14:22-30.
15. Zhang WJ, Wei H, Tien YT, et al. Genetic ablation of phagocytic NADPH oxidase in mice limits TNFα-induced inflammation in the lungs but not other tissues. Free Radic Biol Med, 2011, 50:1517-1525.

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The Regulation of GSTM5 in Oxidative Stress Induced by TNF-α in Human Bronchus Epithelial Cells

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