Vanadium (V) can induce cell apoptosis in layers' oviduct resulting in egg quality reduction. In this study, we investigated the relationship between the mitogen‐activated protein kinase (MAPK)‐signaling pathway and V‐induced apoptosis in poultry oviduct magnum epithelial cells (OMECs). Cultured OMECs were divided into 8 treatment groups: 0 μmol/L V (control), 100 μmol/L V (V100), V100 + P38MAPK inhibitor (SB203580), SB203580, V100 + extracellular signal‐regulated kinases 1 and 2 (ERK1/2) inhibitor (U0126), U0126, V100 + c‐JUN NH2‐terminal kinase (JNK) inhibitor (SP600125), and SP600125. The OMECs were pretreated with the MAPK inhibitors before their treatment with V100 for 12 h. V100 increased the apoptosis of OMECs (P < .05), while 3 MAPK inhibitors suppressed V100‐induced apoptosis P < .05); V100 enhanced the depolarization of △ψm (P < .05), and SB203580 and U0126 alleviated the V100‐induced △ψm decrease (P < .05); V100 downregulated B‐cell lymphoma‐2 (Bcl‐2) and poly [Adenosine diphosphate ribose] polymerase 1 (PARP1) mRNA expression (P < .05), meanwhile it upregulated Bcl‐2 associated x (Bax), Apaf1, cytochrome C (CytC) and cysteine aspartase (caspase) 3, 8, 9 mRNA expression (P < .05). All MAPKs inhibitors alleviated the up‐regulation of V100 for Bax and caspase 3 mRNA expression and down‐regulation of V100 for Bcl‐2 expression (P < .05). SB203580 and U0126 upregulated CytC expression treated by V100 (P < .05), except SP600125, while SB203580 administration resulted in a similar upregulation of PARP1 expression (P < .05). SP600125 can alleviated V triggered p‐P38MAPK (phosphor‐P38), p‐ERK1/2 (phosphor‐ERK1/2), p‐JNK (phosphor‐JNK) increase on OME cells, and SB203580 and U0126 had a similar response to phosphor‐P38 and p‐JNK (P < .05). It concluded that V‐induced apoptosis in OMECs through the activation of P38 and ERK1/2, and by increasing the ratio of Bax/Bcl‐2, which resulted in △ψm decrease, CytC release into the cytosol; consequently caspase 3 is recruited and activated, PARP1 is cleaved, eventually leading to apoptosis.