Objective To elucidate the effect of adipose derived stem cell (ADSC) on mitochondrial membrane permeability transition pore (MPTP) in rats with hepatic ischemia‑reperfusion (IR) injury and the possible mitochondrial protective mechanisms. Methods Two female SD rats were used for isolation, culture, and identification of ADSC. According to the random number table method, 24 SD rats were divided into four groups (n=6): a sham operation (Sham) group, an IR group, an IR+ADSC group, and an IR+ADSC+MPTP opener [carboxyatractyloside (CATR)] group. In the Sham group, rats underwent laparotomy without ischemia treatment. In the IR and IR+ADSC groups, 50 µl phosphate buffer saline (PBS) or 50 µl (5×10⁵ cells/µl) ADSC suspension was injected via the portal vein immediately before ischemia, respectively. In the IR+ADSC+CATR group, rats were injected with 5 mg/kg CATR via the portal vein 30 min before ischemia, based on the treatment in the IR+ADSC group. A rat model of 70% hepatic warm IR injury was established through ischemia over 60 min, followed by reperfusion for 6 h and then serum and liver tissue samples were collected. The changes in serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels were measured. Liver necrosis areas and the percentage of hepatocyte apoptosis were evaluated by hematoxylin‑eosin (H‑E) staining and terminal deoxynucleotidyl transferase‑mediated dUTP‑biotin nick end labeling assay (TUNEL). The expression of extracellular signal‑regulated kinase 1/2 (ERK1/2), phosphorylated ERK1/2 (p‑ERK1/2), glycogen synthase kinase‑3β (GSK‑3β), and phosphorylated GSK‑3β (p‑GSK‑3β) was detected by immunohistochemistry and Western blot. The levels of reactive oxygen species (ROS), superoxide dismutase (SOD), malondialdehyde (MDA), and cyclic adenosine monophosphate (cAMP) in liver tissue were measured. The hepatic mitochondria were isolated to detect mitochondrial calcium retention capacity (CRC). Results The positive expression rates of surface markers CD90, CD73, and CD44 on the third‑generation ADSC were 96.9%, 90.6%, and 93.3%, respectively, while the expression rates of CD45, CD34, and CD11b were all below 1%. The adipogenic, osteogenic, and chondrogenic differentiation assays indicated that the isolated ADSC had multilineage differentiation potential and could be successfully induced to differentiate into adipocytes, osteoblasts, and chondrocytes. H‑E staining revealed focal necrosis, severe vacuolar degeneration, and extensive inflammatory cell infiltration in the liver tissue of the IR group, while vacuolar degeneration and necrosis were significantly reduced in the IR+ADSC group. TUNEL results showed increased apoptotic hepatocytes in the IR group but decreased apoptosis in the IR+ADSC group. Compared with the Sham group, the IR, IR+ADSC, and IR+ADSC+CATR groups exhibited increased necrotic areas in the liver (all P<0.05), increased TUNEL‑positive cell proportions, elevated ALT and AST levels, increased ROS, MDA and cAMP levels, and increased p‑ERK1/2/ERK1/2 ratios (all P<0.05), along with reduced SOD levels (all P<0.05). Mitochondrial CRC was significantly reduced in the IR and IR+ADSC+CATR groups (all P<0.05); the p‑GSK‑3β/GSK‑3β ratio decreased in the IR group (P<0.05), but increased in the IR+ADSC group (P<0.05). Compared with the IR group, the IR+ADSC group showed reduced necrotic areas in the liver, decreased TUNEL‑positive cell proportions, reduced ALT and AST levels, and reduced ROS and MDA levels (all P<0.05), along with increased cAMP levels, p‑ERK1/2/ERK1/2 ratios, SOD levels, and mitochondrial CRC (all P<0.05). The p‑GSK‑3β/GSK‑3β ratio decreased in the IR+ADSC group, the IR+ADSC+CATR group (all P<0.05). Compared with the IR+ADSC group, the IR+ADSC+CATR group showed increases in the necrotic areas of the liver (P<0.05), increases in TUNEL‑positive cell proportions, ALT and AST levels, and ROS levels (all P<0.05), along with decreases in mitochondrial CRC (P<0.05). There were no statistical differences in other indicator among the groups (all P>0.05). Conclusions ADSC pretreatment can inhibit MPTP opening during hepatic ischemia reperfusion, thereby reducing hepatocyte necrosis and apoptosis. The underlying mechanism is related to the regulatory role of ADSC in the cAMP‑ERK1/2‑GSK‑3β signaling pathway.