Nitiric oxide (NO) and eicosanoids function as crucial immune mediators in insects. This study focuses on cross-talkbetween two immune mediators in term of humoral immune response in Spodoptera exigua. Up-regulation of eight differentanti-microbial peptides (AMP) was observed against bacterial challenge. In contrast, injection of E. coli along with L-NAMEsignificantly down regulation the AMP production whereas D-NAME was not effective. The injection of E. coli withdexamethasone or SNAP also decreased AMP production whereas arachidonic acid (AA) compensated the dexamethasoneeffects. RNA inerferece against SeNOS showed the down-regulation of defensin mRNA level, whereas dsNOS injectionswith AA reversed the gene regulation.