To study the effects of different concentration of glycerol on the sperm motility and related enzymes in the frozen bovine semen dilution. Three groups of different concentrations of glycerol (A set of 5.0%, B set of 7.0%, C set of 9.0%) were compared. The results showed that: the osmotic pressure of different concentrations glycerol were 1196.4 ± 4.16, 1532.2 ± 7.19, 1733.4 ± 2.88 mmol/kg. The difference between the groups was significant (P <0.05). It demonstrated that the added amount of glycerol of dilution could cause osmotic pressure rises. The motility of A group was higher than B and C groups(P <0.05), after the diluted and balanced of semen. The aspartate aminotransferase (AST) seepage quantity of A group was lower than B and C groups (P <0.05). The lactate dehydrogenase catalase (LDH) seepage quantity of A group was lower than B and C groups(P <0.05). The motility of A group after freezing was higher than B and C groups(P <0.05). The aspartate aminotransferase (AST) seepage quantity of A group was lower than B and C(P <0.05)groups The lactate dehydrogenase catalase (LDH) seepage quantity of A group was lower than B and C groups (P <0.05).The effects of high concentration of glycerol on sperm motility was mainly caused by the change of osmotic pressure. The seepage quantity of AST, LDH could testify to the extent of damage of sperm cells (on the sperm motility). The concentration for 5.0% glycerol of diluent was the best.%为探讨牛冷冻精液稀释液中不同甘油浓度对精子活力及相关酶渗出量的影响，以不同甘油浓度（A组5.0％、B组7.0％、C组9.0％）进行了试验比较。结果表明：A、B、C组的渗透压值分别为1196.4±4.16mmol/kg、1532.2±7.19mmol/kg、1733.4±2.88mmol/kg，组间差异显著（P<0.05），证实稀释液中甘油量的增加可引起渗透压的升高。精液经稀释平衡后，精子活力A组高于B、C组（P<0.05）；精清中谷草转氨酶（AST）的渗出量，A组低于B、C组（P<0.05）；乳酸脱氢酶（LDH）的渗出量A组低于B、C组（P<0.05），B组与C组差异不显著（P>0.05）。冷冻后，精子活力A组高于B、C组（P<0.05）；精清中谷草转氨酶（AST）的渗出量A组低于B、C组（P<0.05）；乳酸脱氢酶（LDH）渗出量A组低于B、C组（P<0.05）。试验显示，高浓度甘油对精子活力的影响，主要是渗透压变化所致。AST、LDH渗出量的高、低可印证精液中精子细胞受损程度。5.0％甘油浓度的稀释液为最理想。
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