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Aim: To investigate the antioxidant properties of berberine (Ber) on corpus cavenosum smooth muscle cells (CCSMC) in penile erectile dysfunction. Methods: We examined the effects of Ber on cultured rabbit CCSMC damaged by hydrogen peroxide (H2O2) through examining cell viability by methyl thiazolyl tetrazolium assay and assessing the level of malondialdehyde (MDA), superoxide dismutase (SOD) activity, nitric oxide (NO) products, and lactate dehydrogenase (LDH) re- lease in cells after stimulation with H202. Results: Treatment with 1 mmol/L H2O2 significantly decreased the cell viability, NO products, and SOD activity of CCSMC from 100% to 48.57%+4.1% (P<0.01), 66.8±16.3 to 6.7±2.1 μmol/L (P<0.01), and 49.5±1.8 to 30.1±2.6 U/mL (P<0.01), respectively, and increased LDH release and MDA content from 497.6±69.5 to 1100.5±56.3 U/L (P<0.01) and 3.7±1.3 to 78.4±2.9 nmol/mg protein (P<0.01), respectively. However, treatment with different con- centrations of Ber (10-1000 μmol/L) inhibited the damaging effects of H2O2, with increased cell viability (P<0.05 or P<0.01), NO production (P<0.01), and SOD activity (P<0.01) and decreased LDH release and MDA content (both P<0.01). Conclusion: Bet could produce its antioxidant action on oxidative stress-induced cultured CCSMC. These effects may be of benefit in the prevention of penile erectile dysfunction.