Notch signaling is involved in regulation of LPS-induced macrophage apoptosis through JNK/NF-kB signaling pathway

Abstract

Notch signaling plays a critical role in the development and function of macrophages. The aim of the present study was to investigate the relationship between Notch signaling pathway and macrophage apoptosis after LPS stimulation. In RAW 264.7 cells, the mRNA expression of Jagged1, Hes1, Hes 5 and GM-CSF, and protein expression of NICD1 and GM-CSF were increased after LPS stimulation. Inhibition of Notch signaling by γ-secretase inhibitor DAPT and the suppression of Notch1 expression using siRNA both significantly prevented LPS induced activation of JNK and NF-kB, and simultaneously the expression of GM-CSF was also down regulated significantly. JNK inhibitor SP600125 was used to block the phosphorylation of JNK signaling, Western blot results showed that the activation of NF-kB was blocked and expression of GM-CSF was down-regulated. Finally, flow cytometry analyses showed that the Notch signaling was involved in the regulation of macrophage apoptosis after LPS stimulation. Our study showed that the Notch signaling pathway was activated and involved in the regulation of macrophage apoptosis after LPS stimulation through JNK/ NF-kB signaling regulated GM-CSF expression.