Mitochondrial dysfunction is associated with type 2 diabetes mellitus (T2
DM
). 5‐Aminolevulinic acid (
ALA
), a natural amino acid produced only in the mitochondria, is a precursor of heme. Cytochromes that contain heme play an important role in aerobic energy metabolism. Thus,
ALA
may help reduce T2
DM
‐associated hyperglycemia. In this study, we investigated the effect of
ALA
combined with sodium ferrous citrate (
SFC
) on hyperglycemia in Zucker diabetic fatty (
ZDF
) rats. We found that the gavage administration of
ALA
combined with
SFC
(
ALA
/
SFC
) for 6 weeks reduced plasma glucose and hemoglobin A1c (HbA1c) levels in rats without affecting plasma insulin levels. The glucose‐lowering effect depended on the amount of
ALA
/
SFC
administered per day. Furthermore, the glucose tolerance was also significantly improved by
ALA
/
SFC
administration. Although food intake was slightly reduced in the rats administered
ALA
/
SFC
, there was no effect on their body weight. Importantly,
ALA
/
SFC
administration induced heme oxygenase‐1 (
HO
‐1) expression in white adipose tissue and liver, and the induced expression levels of
HO
‐1 correlated with the glucose‐lowering effects of
ALA
/
SFC
. Taken together, these results suggest that
ALA
combined with ferrous ion is effective in reducing hyperglycemia of T2
DM
without affecting plasma insulin levels.
HO
‐1 induction may be involved in the mechanisms underlying the glucose‐lowering effect of
ALA
/
SFC
.