Objective: To investigate the molecular mechanism of mitochondrial unfolded protein reaction (mtUPR) induced by down-regulation of Drp1 expression in MEF cells. Methods: The expression levels of CHOP, ABCB10, LONP1, and Hsp60 were detected by Western blotting in Drp1 KO/KD cells, MEF cells treated with Drp1 inhibitors Mdivi1 or P110 and control cells after administration of 3-NP at different concentrations (0, 2.5, 5, 10 mM). RT-qPCR was used to detect ABCB10 mRNA levels in Drp1 KD or Mdivi-1 treated MEF cells. Results: The expression levels of CHOP were upregulated in Drp1 KO/KD cells or MEF cells treated with Drp1 inhibitors Mdivi-1 or P110. The expression levels of ABCB10 and mitochondrial unfolded protein response related proteins LONP1 and Hsp60 were upregulated in Drp1 KO/KD cells or MEF cells treated with Drp1 inhibitors Mdivi-1 or P110. Compared with control groups, the mRNA levels of ABCB10 were upregulated in Drp1 KD or Mdivi-1 treated MEF cells. Conclusions: Deficiency of Drp1 activates the mitochondrial unfolded protein response through ABCB10, which causes the upregulation of the mitochondrial unfolded protein reaction proteins CHOP, LONP1 and Hsp60.