Iron deficiency anemia (IDA) is a global public health problem that imposes negative effects on human quality of life, especially in pregnant women and children. Silkworm feces extract (SFE) has been used for improving renal anemia of maintenance hemodialysis patients. In this study, we aimed to explore the preventive efficacy of SFE toward iron deficiency anemia in rats and clarify the underlying mechanisms. Our study demonstrated that SFE treatment improved hematological parameters, decreased TIBC, increased serum iron, transferrin saturation, liver iron and spleen iron of IDA rats. Western blot analysis showed that SFE upregulated iron-regulatory proteins (IRPs) expression and downregulated ferritin expression in livers. Furthermore, SFE inhibited hepcidin expression and increased ferroportin1 expression by blocking JAK2/STAT3, BMP6/SMAD, and HFE/TfR2 pathways. And SFE also attenuated hepcidin expression induced by IL-6 and BMP6 in HepG2 cells. These findings suggested that SFE may be an efficient oral iron supplement to ameliorate iron deficiency anemia.