In this study, the effects of adenoviral-mediated glial cell line-derived neurotrophic factor (Ad-GDNF) on apoptosis of cultured bone marrow mesenchymal stem cells (BMSCs) induced by hydrogen peroxide (H2O2) were investigated. After BMSCs infected with Ad-GDNF were treated with 500 μM H2O2 at 37° C for 1 h, lactate dehydrogenase (LDH), MTT and TUNEL methods were used to detect cell viability and apoptosis. In addition, the levels of GDNF in the supernatants in BMSCs cultures were detected by ELISA, pro-inflammatory cytokines IL-6 and TNF-a in the supernatants and apoptosis-related protein Bax and Bcl-2 in cell pellets were investigated. The results showed that H2O2 treatment significantly induced apoptosis of BMSCs and decreased the viability of BMSCs. However, Ad-GDNF significantly reversed the effects of H2O2 on BMSCs. Furthermore, Ad-GDNF significantly decreased the levels of IL-6 and TNF-a and increased Bcl-2/Bax ratio in BMSCs treated by H2O2. In conclusion, Ad-GDNF inhibits apoptosis of BMSCs induced by H2O2, and the mechanisms may be related to down-regulating the expression of IL-6 and TNF-a and up-regulating the ratio of Bcl-2/Bax in BMSCs.