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AIM: To investigate the effect of berberine on proliferation of <i>in vitro</i> pterygium fibroblasts and its possible mechanism.<p>METHODS: Pterygium fibroblast obtained from cultured pterygium tissue after surgical excision. Pterygium fibroblasts cells apoptosis level, mitochondrial membrane potential and apoptosis-related factors mRNA and protein expression levels were detected after the induction of berberine with different final concentrations(0, 20, 40, 80μmol/L).<p>RESULTS: Berberine increased the mitochondrial depolarization level, apoptosis rate, expression level of pro-apoptotic gene Bax, Bad mRNA and protein, and decreased the expression level of Bcl-2 gene mRNA and protein <i>in vitro</i> pterygium fibroblasts cells in a dose-dependent manner. <p>CONCLUSION: Berberine may induce <i>in vitro</i> cultured pterygium cell apoptosis by increasing mitochondrial depolarization.