ABSTRACT.Purpose: The goal of the present study was to investigate whether the antioxidants vitamin E, vitamin C and vitamin B1 can reduce the transforming growth factor-beta2 (TGF-b2)-induced gene expressions in cultured human optic nerve head (ONH) astrocytes. Methods: Cultured human ONH astrocytes were pretreated with different concentrations of vitamin E, vitamin C and vitamin B1 and then exposed to 1.0 ng ⁄ ml TGF-b2 for 24 hr. Expression of the heat shock proteins Hsp27 and aB-crystallin, the extracellular matrix (ECM) component fibronectin and the ECM-modulating protein connective tissue growth factor (CTGF) was detected by immunohistochemistry or real-time PCR analysis. Results: TGF-b2 increased the expression of Hsp27, aB-crystallin, fibronectin and CTGF in human ONH astrocytes. Pretreatment with different concentrations of vitamin E, vitamin C and vitamin B1 reduced the TGF-b2-stimulated gene expressions. Conclusion: In cultured human ONH astrocytes, the TGF-b2-stimulated gene expressions could be reduced by pretreatment with vitamin E, vitamin C and vitamin B1. Therefore, the use of antioxidants in glaucomatous optic neuropathy might be a promising approach to prevent TGF-b2-induced cellular changes in ONH astrocytes.