Purpose: The spleen is a crucial organ manifesting immune functions. Thus, radiation-induced oxidative challenge is vulnerable for the spleen. Our major objective was to protect the spleen from radiation-induced anomalous situations and to identify the signaling pathways involved. Materials and methods: Swiss albino mice were treated with ferulic acid (FA) once in a day at a dose of 50 mg/kg body weight for 5 consecutive days before exposing them to single dose of 10 Gy irradiation. The ROS generation and MMP change were determined by flow cytometry. The expression of different signaling proteins was investigated by immunoblotting and immunocytochemistry. Results: FA pretreatment significantly prevented radiation-induced oxidative stress by downregulating TBARS formation and by upregulating SOD and catalase activity. FA scavenged ROS, prevented the alteration of MMP and downregulated the expression of stress marker Cdc42 and apoptotic markers p53, p21, Bax and PTEN. Cell cycle analysis showed DNA damage induced arrest of cells at subG0/G1 phase. Moreover, pretreatment with FA augmented Bcl2 expression and also increased the level of p-PI3K. Conclusion: FA prevented the activation of apoptotic signaling events in the spleen by interfering with the free radical chain reaction and by scavenging superfluous ROS. This is perhaps the first comprehensive study with a mechanistic viewpoint that FA can protect the spleen from ionizing radiation. © 2016 Informa UK Limited, trading as Taylor & Francis Group.