Abstract: AIM:To investigate whether adiponectin (APN) generates cardioprotection in response to high-glucose/high-fat damage through increasing mitochondrial biogenesis and functions. METHODS: Cultured neonatal rat ventricular myocytes (NRVM) were randomized into control group (5 mmol/L glucose and 20 mmol/L mannitol), high-glucose/high-fat group (25 mmol/L glucose and 500 μmol/L sodium palmitate, 18 h incubation) and high-glucose/high-fat+APN group (25 mmol/L glucose, 500 μmol/L sodium palmitate and 3 μg/ml globular APN, 18 h incubation) after 3 days of normal culture. Cells were harvested for evaluation of mitochondrial membrane potential and mRNA levels of Tfam after high-glucose/high-fat incubation. RESULTS: Compared with those in control group, both mitochondrial membrane potential and mRNA levels of Tfam decreased and cell apoptosis increased in high-glucose/high-fat group. However, these changes were reversed by APN. CONCLUSION: High-glucose/high-fat can lead to mitochondrial dysfunction in cardiomyocytes and APN can protect cardiomyocytes by increasing mitochondrial biogenesis and functions.