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目的探讨不同精卵共孵育时间对小鼠体外受精受精率、氧化应激产生及胚胎质量的影响,及小鼠体外受精最短的精卵共孵育时间。方法分别比较ICR小鼠精卵共孵育30s、2min、10min、0.5h、2h、4h及18h的体外受精的受精率及囊胚形成率,并对精卵共孵育0h、2h及18h培养液的丙二醛浓度进行检测。结果各组获得卵细胞数分别为52、53、53、46、48、48及47,受精率分别为67.3%、81.1%、83.0%、87.0%、85.4%、81.3%及48.9%。精卵共孵育30s组的受精率显著低于0.5h及2h组(P<0.05),而18h组的受精率显著低于2min、10min、0.5h、2h及4h组(P<0.05)。2min组的受精率相比10min、0.5h、2h及4h组均无差异(P>0.05)。各组的囊胚形成率分别为82.9%、65.1%、70.5%、67.5%、75.6%、71.8%及65.2%,两两比较差异均无显著性(P>0.05)。精卵共孵育0h、2h及18h丙二醛浓度检测的均值分别为(1.06±0.21)×10-3mol/L、(1.44±0.22)×10-3mol/L及(2.00±0.21)×10-3mol/L,各组间两两比较差异均有显著性(P<0.05),丙二醛浓度随着精卵共孵育时间的缩短而降低。结论小鼠体外受精精卵共孵育时间缩短至2min不会降低受精率,同时随着孵育时间的缩短,产生的氧化应激也随之减少。
Objective To investigate the effect of co-incubation time of different spermatozoa on in vitro fertilization rate, oxidative stress and embryo quality in mice, and the incubation time of the sperm in mice with the shortest in vitro fertilization. Methods The fertilization rate and blastocyst formation rate of in vitro fertilization of ICR mouse sperm incubated for 30s, 2min, 10min, 0.5h, 2h, 4h and 18h were compared respectively, and incubated for 0h, 2h and 18h Malondialdehyde concentration was tested. Results The numbers of oocytes obtained in each group were 52, 53, 53, 46, 48, 48 and 47, respectively. The fertilization rates were 67.3%, 81.1%, 83.0%, 87.0%, 85.4%, 81.3% and 48.9%, respectively. The sperm fertilization rate in 30s group was significantly lower than that in 0.5h and 2h groups (P <0.05), while the fertilization rate in 18h group was significantly lower than that in 2min, 10min, 0.5h, 2h and 4h groups (P <0.05). The fertilization rate in 2min group was no significant difference compared with 10min, 0.5h, 2h and 4h groups (P> 0.05). The blastocyst formation rates of each group were 82.9%, 65.1%, 70.5%, 67.5%, 75.6%, 71.8% and 65.2%, respectively. There was no significant difference between the groups (P> 0.05). The mean concentrations of malondialdehyde (MDA) detected at 0h, 2h and 18h were (1.06 ± 0.21) × 10-3mol / L, (1.44 ± 0.22) × 10-3mol / L and (2.00 ± 0.21) × 10- 3mol / L, there was a significant difference between every two groups (P <0.05). The concentration of malondialdehyde decreased with the time of sperm incubation. Conclusions The short incubation time of in vitro fertilized sperm in mice can not reduce the fertilization rate, and the oxidative stress also decreases with the shortening of incubation time.