Cigarette smoke extract (CSE) induces transient receptor potential ankyrin 1(TRPA1) expression via activation of HIFI cdn A549 cells

摘要

We previously found that transient receptor potential ankyrin 1 (TRPA1) in guinea pig tracheal epithelial cells was elevated after 14 days of cigarette smoke (CS) exposure. However, the mechanism underlying CS-induced TRPA1 expression remains unknown. Here, we explored whether cigarette smoke extract (CSE)-induced TRPA1 expression is related with modulation of HIF1 alpha in A549 cells. Our results showed that CSE increased TRPA1 expression in A549 cells, decreased I kappa B, PHD2, and HDAC2, and increased ROS release and nuclear translocation of NF-kappa B and HIF1 alpha. Moreover, HIF1 alpha siRNA and/or MG132 (a proteasome inhibitor) pretreatment significantly inhibited CSE-induced TRPA1 expression and HIF1 alpha nuclear translocation in A549 cells. However, HIFloc siRNA pretreatment did not affect CSE-induced NF-kappa B nuclear translocation, suggesting that CSE-induced TRPA1 expression in A549 cells is directly mediated by HIFloc, but not by NF-kappa B. Similar to CSE treatment, treatment of A549 cells with LPS caused significant increases in nuclear translocation of NF-kappa B and HIFloc mRNA expression, but did not alter TRPA1 mRNA expression. However, pretreatment with PHD2 siRNA did result in increased TRPA1 mRNA expression in LPS-treated A549 cells; an effect that was inhibited by SN50 (a NF-kappa B inhibitor). It suggests a role for NF-kappa B to indirectly regulate TRPA1 mRNA expression via modulating HIFloc mRNA transcription. In addition, treatment cells with HDAC2 siRNA plus 2%CSE resulted in increased HIFI oc nuclear trans location and TRPA1 expression, which was significantly inhibited by MG132 and HIFI oc siRNA. These results suggest that HDAC2 indirectly modulates TRPA1 expression by promoting the DNA-binding activity of HIFloc. These findings show that CSE increases TRPA1 expression in airway epithelial cells by directly activating HIFloc, and that this increase in TRPA1 expression is indirectly regulated via NF-kappa B, PHD2 and HDAC2 modulation of HIFI oc activity. (C) 2016 Published by Elsevier Inc.