GSTM5 Down-regulates TNF-α-Mediated Inflammation Levels in Human Bronchial Epithelial Cells by p38MAPK/NF-κB Pathway_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

HuangYun ¹ , YuanWeifeng ² , LiLi ² ,  HuangWenjie ²

1. Postgraduate Institute of Guangzhou Medical University, Guangzhou, Guangdong, 510182, China;
2. ;

Corresponding author：

HuangWenjie, Email: huangyelu1029@vip.163.com

Keywords：

Glutathione S-transferase M5; Acute lung injury; Inflammation cytokines; Nuclear factor-κB; p38 mitogen-activated protein kinase

DOI：

10.7507/1671-6205.2016040

Video：

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Objective To investigate the effects of glutathione S-transferase M5 (GSTM5) on the inflammation in human bronchial epithelial 16HBE cells and its possible molecular mechanisms. Methods Acute lung injury cell model was constructed with 16HBE cells induced by tumour necrosis factorα (TNF-α, 10 ng/mL). The cells were devided into a control group, a TNF-α group (TNF-α), a GSTM5 group (GSTM5+TNF-α), a negative control group (negative control plasmid+TNF-α). GSTM5-GFP plasmid and negative control plasmid were respectively transfected to the cells of the GSTM5 group and the negative control group using Lipofectamine2000. The contents of interleukin-6(IL-6), IL-8, IL-10 in the cell supernatant were measured by ELISA.The expression of nuclear factor-κB (NF-κB) mRNA was detected by RT-PCR, and the expression of NF-κB, phospho-NF-κB, p38, phospho-p38 protein were detected by Western blot. Results The GSTM5-GFP eukaryotic expression vector was successfully constructed and transfected successfully confirmed by fluorescence microscope. The contents of IL-6, IL-8, IL-10 in the TNF-α-induced cell supernatant were significantly higher than those in the control group(P < 0.05), and the contents of IL-6, IL-8, IL-10 in the GSTM5 group were lower than those in the TNF-α group (P < 0.05)with statistically significant difference. At the same time, the total NF-κB mRNA, phospho -NF-κB and phospho-p38 protein were increased in TNF-α stimulated cells compared with the control group (P < 0.05), while the GSTM5 group was lower than that in the TNF-α group and the negative control group (P < 0.05). Conclusion Overexpression of GSTM5 inhibits the phosphorylation of p38MAPK and NF-κB and down-regulates the inflammation of TNF-α-induced human bronchial epithelial 16HBE cells.

Citation： HuangYun, YuanWeifeng, LiLi, HuangWenjie. GSTM5 Down-regulates TNF-α-Mediated Inflammation Levels in Human Bronchial Epithelial Cells by p38MAPK/NF-κB Pathway. Chinese Journal of Respiratory and Critical Care Medicine, 2016, 15(2): 171-175. doi: 10.7507/1671-6205.2016040 Copy

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1. Cross LJ, Matthy MA. Biomarkers in acute lung injury: insights into the pathogenesis of acute lung injury. Crit Care Clin, 2011, 27:355-377.
2. Zivkovi M, Stankovi A, Djuri T, et al. Effects of glutathione S-transferase T1 and M1 deletions on advanced carotid atherosclerosis, oxidative, lipid and inflammatory parameters. Mol Biol Rep, 2014, 41:1157-1164.
3. 吴炜景, 李理, 袁伟锋, 等.NF-κB p65基因在TNF-α诱导的肺上皮细胞氧化损伤中的作用.中国呼吸与危重监护杂志, 2012, 11:46-51.
4. 杜鹃, 高伟良, 冯清州, 等.p38MAPK信号通路在LPS诱导RAW264.7细胞表达sTREM-1中的作用.中国呼吸与危重监护杂志, 2015, 14:161-164.
5. Matthay MA. Treatment of acute lung injury: clinical and experimental studies. Proc Am Thorac Soc, 2008, 5:297-299.
6. Hu XY, Neill SJ, Cai WM, et al. Induction of defence gene expression by oligogalacturonic acid requires increases in both cytosolic calcium and hydrogen peroxide in Arabidopsis thaliana. Cell Res, 2004, 14:234-240.
7. Uno Y, Murayama N, Kunori M, et al. Systematic identification and characterization of glutathione S-transferases in cynomolgus macaque. Biochem Pharmacol, 2013, 86:679-690.
8. 刘涛, 任成山.炎症介质在急性肺损伤/急性呼吸窘迫综合征发病机制中的作用.中华肺部疾病杂志(电子版), 2013, 6:265-269.
9. Xu M, Cao F, Liu L, et al. Tanshinone IIA-induced attenuation of lung injury in endotoxemic mice is associated with reduction of hypoxia-inducible factor 1α expression. Am J Respir Cell Mol Biol, 2011, 45:1028-1035.
10. Yu PJ, Li JR, Zhu ZG, et al. Praeruptorin D and E attenuate lipopolysaccharide/hydrochloric acid induced acute lung injury in mice. Eur J Pharmacol, 2013, 710:39-48.
11. Chen X, Yang X, Liu T, et al. Kaempferol regulates MAPKs and NF-κB signaling pathways to attenuate LPS-induced acute lung injury in mice. Int Immunopharmacol, 2012, 14:209-216.
12. Yang R, Yang L, Shen X, et al. Suppression of NF-κB pathway by crocetin contributes to attenuation of lipopolysaccharide-induced acute lung injury in mice. Eur J Pharmacol, 2012, 674:391-396.
13. Keshet Y, Seger R. The MAP kinase signaling: a system of components regulates a diverse array of physiological functions. Methods Mol Biol, 2010, 661:3-38.

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GSTM5 Down-regulates TNF-α-Mediated Inflammation Levels in Human Bronchial Epithelial Cells by p38MAPK/NF-κB Pathway

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