Abstract
BX-795 is an inhibitor of 3-phosphoinositide-dependent kinase 1 (PDK1), but also a potent inhibitor of the IKK-related kinase, TANKbinding kinase 1 (TBK1) and IKKɛ. In this study we attempted to elucidate the molecular mechanism(s) underlying the inhibition of BX-795 on Herpes simplex virus (HSV) replication. HEC-1-A or Vero cells were treated with BX-795 and infected with HSV-1 or HSV-2 for different periods. BX-795 (3.125-25 μmol/L) dose-dependently suppressed HSV-2 replication, and displayed a low cytotoxicity to the host cells. BX-795 treatment dose-dependently suppressed the expression of two HSV immediate-early (IE) genes (ICP0 and ICP27) and the late gene (gD) at 12 h postinfection. HSV-2 infection resulted in the activation of PI3K and Akt in the host cells, and BX-795 treatment inhibited HSV-2-induced Akt phosphorylation and activation. However, the blockage of PI3K/Akt/mTOR with LY294002 and rapamycin did not affect HSV-2 replication. HSV-2 infection increased the phosphorylation of JNK and p38, and reduced ERK phosphorylation at 8 h postinfection in the host cells; BX-795 treatment inhibited HSV-2-induced activation of JNK and p38 MAP kinase as well as the phosphorylation of c-Jun and ATF-2, the downstream targets of JNK and p38 MAP kinase. Furthermore, SB203580 (a p38 inhibitor) or SP600125 (a JNK inhibitor) dose-dependently inhibited the viral replication in the host cells, whereas PD98059 (an ERK inhibitor) was not effective. Moreover, BX-795 blocked PMA-stimulated c-Jun activation as well as HSV-2-mediated c-Jun nuclear translocation. BX-795 dose-dependently inhibited HSV-2, PMA, TNF-α-stimulated AP-1 activation, but not HSV-induced NF-κB activation. Overexpression of p38/JNK attenuated the inhibitory effect of BX-795 on HSV replication. BX-795 completely blocked HSV-2-induced MKK4 phosphorylation, suggesting that BX-795 acting upstream of JNK and p38 MAP kinase. In conclusion, this study identifies the anti-HSV activity of BX-795 and its targeting of the JNK/p38 MAP kinase pathways in host cells.
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Acknowledgements
We thank Dr Qi-han LI at the Institute of Medical Biology, Chinese Academy of Medical Sciences for HSV-1 (HF), Dr Er-guang LI at the School of Medicine, Nanjing University, China for HSV-2 (G) and Dr Claus-Henning NAGEL at the Heinrich Pette Institute-Leibniz Institute for Experimental Virology, Germany for pcDNA3-ICP0-1/2-GFP.
This study was supported by the Major Research and Development Project from the Ministry of Health (Grant No 2013ZX10001005-003 and 2016ZX10001005003), the National Key Research and Development Program of China (Grant No 2016YFC1201000), Jiangsu Natural Science Foundation (Grant No BK20130591), MOE Doctoral Base Foundation (Grant No 20130091120022), State Key Laboratory of Analytical Chemistry for Life Science (Grant No 5431ZZXM1615).
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Su, Ar., Qiu, M., Li, Yl. et al. BX-795 inhibits HSV-1 and HSV-2 replication by blocking the JNK/p38 pathways without interfering with PDK1 activity in host cells. Acta Pharmacol Sin 38, 402–414 (2017). https://doi.org/10.1038/aps.2016.160
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DOI: https://doi.org/10.1038/aps.2016.160
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