Abstract: Cryopreservation of stallion semen has not reached the level of efficiency and positive results like other domestic farm animals. This is mainly due to the greater sensitivity of stallion sperm to the freezing process, showing higher rates of oxidative stress and plasma membrane damage, which suggest a requirement for the methods of improving the efficiency of this technique in stallion. In this experiment, Glutathione（Gsh）, taurine（Tau）, ascorbic acid（vitamin C）, methionine（Met） and proanthocyanidins（OPC） were chosen as antioxidants and added into semen dilution, respectively. The overall viability（TM）, progressive viability（PM）, rapid motility sperm ratio（RAP）, mean path velocity（VAP） and straightness（LIN） of semen after freezing and thawing were measured by sperm analyzer to evaluate the effects of different antioxidants on the cryopreservation of stallion semen. In the second step, taurine was used as the base antioxidant and other antioxidants were added to determine the effect of the combination of antioxidants on the preservation of equine sperm after freezing and thawing. Taurine used as the base antioxidant and some of antioxidants mentioned before were added to stallion semen before centrifugation, before freezing, before freezing and after thawing, respectively, to evaluate the effect of antioxidant combination adding procedures on the preservation of equine sperm after freezing and thawing.The results showed that taurine, vitamin C, methionine, proanthocyanidins were added in the experimental group, and the motility parameters of group 2,3,4,5 equine sperm were improved. Among the different antioxidant combination tests, the antioxidant effect of glutathione and methionine was the best in test group 9, and the indexes of equine sperm motility were higher than those of control group, while the antioxidant indexes of test group 8 supplemented with taurine, glutathione and methionine were mostly low. The motility parameters of equine sperm in group 10 added before centrifugation and group 11 added before freezing were higher than those in group 12 added twice before and after freezing（P＜0.05）. These results indicated that the addition of taurine, vitamin C, methionine, proanthocyanidins in semen diluent was beneficial, but the combination of taurine and other antioxidants could not achieve a good antioxidant effect. It is suitable to add antioxidants before centrifugation or freezing, and adding antioxidants twice before and after freezing may harm sperm.