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Effects of spleen tyrosine kinase on oxidative stress-induced mucous hypersecretion of airway epithelial cells

RAN Dan-hua1, CHEN Ling-xiu1, HUANG Hua-ping2, HAN Zhong2, ZHOU Xiang-dong1,2   

  1. 1.Department of Respiratory Medicine， The Second Affiliated Hospital， Chongqing Medical University, Chongqing 400010， China; 2.Affiliated Hospital of Hainan Medical University, Haikou 570102, China
  • Online:2016-02-28 Published:2016-03-29
  • Supported by:

    National Natural Science Foundation of China, 81370111, 31171346

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Abstract:

Objective To explore the effects of spleen tyrosine kinase (Syk) on oxidative stress-induced airway mucous hypersecretion. Methods Human airway epithelial cells 16HBE were cultured in vitro. The mucous hypersecretion model was established by stimulating cells by H2O2. Intervention factors were Syk siRNA and extracellular signal regulated kinase (ERK1/2) specific inhibitor U0126 and P38 specific inhibitor SB203580 of mitogen-activated protein kinase (MAPK) signaling pathway. The cells were randomly divided into the control group, H2O2 group, H2O2+Syk siRNA group, H2O2+negative siRNA group, H2O2+U0126 group, and H2O2+SB203580 group. The cell viability was detected by MTT assay. Transcription and protein expression of mucin (MUC) 5AC were detected by RT-PCR and ELISA respectively. The protein expression of Syk, phosphorylated ERK1/2 (p-ERK1/2), and phosphorylated P38 (p-P38) were detected by Western blotting. The content and distribution of MUC5AC protein were detected by confocal laser technology. Results Compared with the control group, expressions of Syk, p-ERK1/2, p-P38, and MUC5AC significantly increased after being stimulated by H2O2 (P=0.000). Above effects of H2O2 were significantly inhibited after being treated by Syk siRNA, U0126, and SB203580 (P=0.000). Conclusion Syk involves the oxidative stress induced airway mucous hypersecretion via ERK1/2 and P38 MAPK signaling pathways.

Key words: spleen tyrosine kinase, signaling pathway, MUC5AC