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目的探讨丙烯腈(ACN)诱导氧化应激对大鼠附睾精子质量与鞭毛超微结构的影响。方法 40只雄性SPF级健康SD大鼠按体重随机分为4组,染毒组以12.5、25和50 mg/kg ACN灌胃,对照组给予等体积玉米油。1次/d,6 d/周,连续13周。测精子活性氧(reactive oxygen species,ROS)、丙二醛(methane dicarboxylic aldehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)、精子线粒体膜电位(mitochondrial membrane potential,MMP)和精子质量(0.86%生理盐水中游离精子),电子显微镜下观察精子鞭毛超微结构。结果 50 mg/kg组精子MMP较其他组均降低(P<0.05)。50 mg/kg组精子ROS含量较其他组升高(P<0.05)。染毒组SOD活力较对照组降低(P<0.05)。精子鞭毛超微结构显示,25和50 mg/kg ACN组精子鞭毛外部致密纤维和线粒体结构有损伤。精子密度50 mg/kg组为(3.52±0.46)×10~6/ml,较对照组、12.5和25 mg/kg组[分别为(5.71±0.66)×10~6/ml、(4.95±1.33)×106/ml和(5.99±0.78)×106/ml]均降低(P<0.05);25和50 mg/kg组精子活率分别为(62.18±3.4)%和(66.25±7.23)%,较对照组(83.22±13.86)%降低(P<0.05)。d级精子率50 mg/kg组为(40.76±1.47)%,25 mg/kg组为(37.77±3.41)%,与对照组(27.77±4.85)%和12.5 mg/kg组(31.77±4.85)%比较均升高(P<0.05)。精子平均路径速度对照组高于各ACN组,差异有统计学意义(P<0.05)。直线速度50 mg/kg组较对照组降低(P<0.05)。精子ROS含量与精子MMP呈负相关(r=-0.643,P=0.024),与SOD活力呈负相关(r=-0.713,P=0.009),与精子活力呈负相关(r=-0.629,P=0.028);与精子平均路径速度呈负相关(r=-0.839,P=0.001);与d级精子率呈正相关(r=0.699,P=0.011)。结论 ACN可致附睾内精子ROS含量增高,精子鞭毛内线粒体和外部致密纤维超微结构改变,而使精子活动能力和运动速度降低。
Objective To investigate the effects of oxidative stress induced by acrylonitrile on epididymal sperm quality and ultrastructure of flagellar in rats. Methods Forty male SPF healthy SD rats were randomly divided into four groups according to body weight. The rats in the exposure group were administered orally with 12.5, 25 and 50 mg / kg ACN, while the control group were given the same volume of corn oil. 1 time / d, 6 d / week for 13 weeks. The levels of reactive oxygen species (ROS), methane dicarboxylic aldehyde (MDA), superoxide dismutase (SOD), mitochondrial membrane potential (MMP) and sperm mass 0.86% saline free sperm), observed under the electron microscope sperm flagellum ultrastructure. Results 50 mg / kg sperm MMP decreased compared with other groups (P <0.05). The level of ROS in 50 mg / kg group was higher than that in other groups (P <0.05). The activity of SOD in the exposed group was lower than that in the control group (P <0.05). Ultrastructures of sperm flagella showed that outer dense fibers and mitochondrial structure of sperm flagella were damaged in 25 and 50 mg / kg ACN group. (5.71 ± 0.66) × 10 ~ 6 / ml and (4.95 ± 1.33), respectively, compared with control group, 12.5 and 25 mg / kg group at sperm concentration of 50 mg / kg ) (P <0.05). The sperm motility rates in the 25 and 50 mg / kg groups were (62.18 ± 3.4)% and (66.25 ± 7.23)%, respectively (83.22 ± 13.86)% lower than the control group (P <0.05). Compared with the control group (27.77 ± 4.85)% and the 12.5 mg / kg group (31.77 ± 4.85), the d-sperm sperm count was (40.76 ± 1.47)% in the 50 mg / kg group and (37.77 ± 3.41) % Were higher (P <0.05). The average path velocity of sperm in control group was higher than that of ACN group, the difference was statistically significant (P <0.05). Linear velocity 50 mg / kg group than the control group decreased (P <0.05). There was a negative correlation between sperm motility and sperm motility (r = -0.643, P = 0.024), but negatively correlated with sperm motility (r = -0.713, P = 0.009) (R = -0.839, P = 0.001). There was a positive correlation between sperm motility and sperm motility (r = 0.699, P = 0.011). Conclusion ACN can induce intracisternal sperm ROS content increased, ultrastructural changes of mitochondria and outer dense fibers in sperm flagellum, and sperm motility and exercise speed decreased.