To investigation the effect of arbutin on hypoxia/reoxygenation (H/R)-induced injury of cardiomyocytes (H9C2) through the neurogenic locus notch homolog protein 1(Notch1)/hairy and enhancer of split 1 (Hes1) signaling pathway.

An H/R-induced H9C2 cell injury model was established. The optimal intervention concentration of arbutin was screened by pretreating cells with various concentrations (12.50-200.00 μmol·L^-1). H9C2 cells were divided into a normal group, a model group, experimental-L, M, H groups (25.00, 50.00, and 100.00 μmol·L^-1 arbutin, respectively), and an inhibitor group {100.00 μmol·L^-1 arbutin+10.00 μmol·L^-1 N-[N-(3, 5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT)}. Cells in the normal group were cultured under normal conditions, while H/R models were established in the remaining groups. All treatment groups received arbutin pretreatment at concentrations ranging from 12.50 to 200.00 μmol·L^-1 for 24 hours before modeling. Cell proliferation was detected by the CCK-8 assay, and apoptosis was assessed by TUNEL assay; mitochondrial membrane potential was measured by JC-1; reactive oxygen species (ROS), lactate dehydrogenase (LDH), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), and superoxide dismutase (SOD) levels were detected using kits. Expression of Notch1, Hes1, Bcl-2, Bax, and caspase-3 was detected by Western blot.

Arbutin at concentrations of 25.00–200.00 μmol·L^-1 significantly increased the survival rate of H/R-induced H9C2 cells (P<0.05). Arbutin at 25.00, 50.00, and 100.00 μmol·L^-1 was selected as the low, medium, and high doses for subsequent experiments. The creatine kinase isoenzyme MB (CK-MB) levels in the normal group, model group, experimental-H group, and inhibitor group were (2.45±0.39), (8.72±0.82), (3.56±0.42), and (5.48±0.57) U·mL^-1, respectively; the cTnI levels were (22.58±2.23), (68.34±5.96), (30.12±3.26), and (49.62±5.15) ng·L^-1, respectively; the LDH levels were (53.42±5.44), (163.72±13.41), (65.45±6.57), and (98.33±9.85) U·L^-1, respectively; the relative ROS levels were 1.01±0.12, 3.34±0.35, 1.46±0.18, and 2.24±0.23, respectively; the SOD levels were (96.23±8.12), (24.31±2.24), (78.39±7.93), and (43.78±4.49) U·mg^-1, respectively; the GSH-Px levels were (74.82±7.55), (16.42±1.83), (61.32±6.29), and (35.72±3.38) U·mg^-1, respectively; the MDA levels were (15.45±2.03), (58.33±5.57), (21.56±2.24), and (42.47±4.13) μmol·L^-1, respectively; the apoptosis rates were (2.45±0.39)%, (29.86±3.25)%, (9.86±0.91)%, and (21.34±2.25)%, respectively; the red/green fluorescence intensity ratios were (5.78±0.65)%, (1.42±0.20)%, (4.89±0.54)%, and (2.85±0.33)%, respectively; the Notch1 protein expression levels were 1.27±0.12, 0.34±0.03, 1.05±0.08, and 0.54±0.05, respectively; the Hes1 protein expression levels were 0.95±0.10, 0.25±0.03, 0.81±0.08, and 0.57±0.05, respectively; the Bcl-2 protein expression levels were 1.01±0.11, 0.31±0.03, 0.89±0.08, and 0.61±0.06, respectively; the Bax protein expression levels were 0.45±0.04, 1.34±0.12, 0.61±0.06, and 0.93±0.09, respectively; and the caspase-3 protein expression levels were 0.23±0.03, 0.91±0.09, 0.37±0.04, and 0.64±0.06, respectively. Comparisons between the normal group and the model group, between the model group and the experimental-L, M, H groups, and between the experimental-H group and the inhibitor group all showed statistically significant differences for the above indicators (all P<0.05).

Arbutin alleviates H/R-induced H9C2 cell injury by activating the Notch1/Hes1 pathway and inhibiting oxidative stress.