Experimental study of irisin alleviates house dust mite-induced airway epithelial cells inflammation and apoptosis via the NF-κB and JNK signaling pathways
10.12092/j.issn.1009-2501.2022.10.004
- Author:
Ying LI
1
;
Wei YAO
1
;
Meng WANG
1
;
Zhihong YU
1
;
Yuanqi GONG
1
;
Haibing LAN
1
;
Xiefei QI
1
Author Information
1. Department of Intensive Care Unit, The Second Affiliated Hospital of NanChang University
- Publication Type:Journal Article
- Keywords:
apoptosis;
house dust mite;
inflammation;
irisin;
oxidative stress;
ROS
- From:
Chinese Journal of Clinical Pharmacology and Therapeutics
2022;27(10):1106-1112
- CountryChina
- Language:Chinese
-
Abstract:
AlM: To explore the effects of irisin on house dust mite (HDM)-induced inflammation and apoptosis in human airway epithelial cells. METHODS: The human bronchial epithelial cell (16HBE) were cultured with in RPMI1640 culture medium with 10% of fetal bovine serum. After cells reached 85% confluence, the medium was replaced with serum-free culture medium for 12 h. Then the 16HBE cells were treated with various concentrations of HDM (0, 400, 800, 12 00 U/mL) for 24 h. Reactive oxygen species assay kit was used to detected the intracellular ROS generation. And qPCR was used to measure the interleukin (IL)-6, tumor necrosis factor-alpha (TNF-α) mRNA expression of the HDM-induced 16HBE cell. The cells were pre-treated with or without irisin for 2 h before exposure to various concentration of HDM for 24 h. Then reactive oxygen species assay kit was used to detected the intracellular ROS generation. The IL-6, TNF-α mRNA expression of 16HBE cell were measured by qPCR. Meanwhile, the phosphorylated and total P65 NF-κB and JNK proteins were detected by western blot. The pro-apoptosis protein cleaved-caspase3BAX and the anti-apoptosis protein were also detected by western blot. RESULTS: The quantitative assay showed that intracellular ROS in different concentrations of HDM stimulus group were obviously higher than NC group (P < 0.05). And RT-PCR analysis showed a higher expression level of pro-inflammatory cytokine TNF-α and IL-6 mRNA in different concentrations of HDM than in NC group (P < 0.05). Compared with the HDM group, Irisin significantly decreased the level of intracellular ROS of the 16HBE cells (P < 0.05). The released of the pro-inflammatory cytokine TNF-α and IL-6 mRNA was also decreased in irisin treated 16HBE cells (P < 0.05). And compared with control group, BCL-XL anti-apoptosis protein level was decreased and BAX and c-caspase3 pro-apoptosis protein levels were increased in HDM group (P < 0.05), irisin intervention significantly increased the level of BCL-XL and decreased the levels of BAX and cleaved-caspase 3 (P < 0.05). Compared the control group, phosphorylated P65 NF-κB and JNK protein levels were significantly increased after HDM stimulated (P < 0.05), and irisin intervention decreased the protein levels of phosphorylated P65 NF-κB and JNK (P < 0.05). CONCLUSlON: Irisin can effectively improve the inflammation and apoptosis of HDM-induced 16HBE cells, and this protective effect may be related to its inhibition of NF-κB and JNK MAPK signaling pathways. Irisin may be a potential drug for treating lung inflammation.
