摘要
目的:维生素E有清除自由基、抗氧化作用,观察维生素E对环磷酰胺致小鼠生精功能损害的保护作用。方法:实验于2004-09/11在泰山医学院形态学实验室完成。选用健康昆明种雄性小白鼠24只,体质量23～28g。随机将其分成3组,每组8只,分别为正常对照组、环磷酰胺组、维生素E治疗组。环磷酰胺组、维生素E治疗组小鼠经口灌胃5mg/(kg·d)环磷酰胺;维生素E治疗组小鼠每天给予环磷酰胺处理后4h皮下注射50mg/(kg·d)维生素E;正常对照组小鼠经口灌胃等量生理盐水。各组均连续处理28d,于末次给药后24h,处死全部小鼠。结果评估:①按放免试剂盒说明书分别测定小鼠血清卵泡刺激素、黄体生成素、睾酮。②400倍光镜观察小鼠精子悬液,计数精子,计算精子活率、畸形率、精子活力。③在冰浴条件下将小鼠睾丸组织制成睾丸匀浆测定小鼠睾丸组织超氧化物歧化酶、谷胱甘肽过氧化物酶、过氧化氢酶的活性及丙二醛含量;睾丸组织光镜切片后苏木精-伊红染色观察睾丸组织结构。④光镜观察100个曲细精管的精子发生情况,按抗精子发生效应积分评定法(精子发生正常为0分,15%曲细精管有精子发生障碍为1分,15%～30%曲细精管有精子发生障碍为2分,30%～40%曲细精管有精子发生障碍为3分,40%～60%曲细精管有精子发生障碍为4分,60%～75%曲细精管有精子发生障碍为5分,75%～90%曲细精管有精子发生障碍为6分,90%以上曲细精管有精子发生障碍为7分,90%以上曲细精管有精子发生障碍,且仅有少量精原细胞为8分)评价环磷酰胺和维生素E的不同效应。结果:进入结果分析小鼠24只。①环磷酰胺组血清卵泡刺激素、黄体生成素、睾酮、精子密度、活率、活动力及睾丸组织超氧化物歧化酶、谷胱甘肽过氧化物酶、过氧化氢酶活性均低于正常对照组(P<0.05～0.01),精子畸形率、睾丸组织丙二醛含量高于正常对照组(P<0.01),睾丸组织生精细胞层次减少,出现多核巨细胞、坏死等病变,抗精子效应评分高于正常对照犤(6.96±0.83),0分,P<0.01犦。②维生素E治疗组血清卵泡刺激素、黄体生成素、睾酮、精子密度、活率、活动力、睾丸组织超氧化物歧化酶、谷胱甘肽过氧化物酶、过氧化氢酶活性分别高于环磷酰胺组(P<0.05～0.01),精子畸形率、睾丸组织丙二醛含量低于环磷酰胺组(P<0.01),睾丸组织结构较环磷酰胺组明显恢复,抗精子效应评分低于环磷酰胺组犤(0.70±0.55),(6.96±0.83)分,P<0.01犦。结论:维生素E对环磷酰胺致生精功能损害有明显的保护作用,该作用的发挥可能与维生素E清除氧自由基、抗氧化作用及促进垂体前叶促性腺激素的释放等有关。
AIM： Vitamin E can clean free radical and has anti-oxidation action. To observe the protective effects of vitamin E on the damage of mouse spermatogenes induced by cyclophosphamide. METHODS： The experiment was finished in Department of morphology of Taishan Medical College from September to November 2004. Twenty-four Kunming healthy male mice, body mass 23-28 g were divided randomly into three groups with 8 mice in each group： normal control group, cyclophosphamide group, and vitamin E treatment group. The mice in latter two groups were given cyclophosphamide by gastric perfusion through mouth at a dosage of 5 mg/kg per day. The mice in vitamin E treatment group were given vitamin E by subcutaneous injection at a dose of 50 mg/kg per day after 4 hours cyclophosphamide treatment.The mice in normal control group were gavaged with equivalent saline through mouth. The treatment period for all groups was 28 days. All mice were sentenced to death 24 hours after the last treatment. Result evaluation： ① The blood serum of mice was collected to detect the level of plasma follicle stimulating hormone （FSH）, luteinizing hormone （LH）, and testosterone （T）. ② To observe sperm liquor, to count sperm quantity, to calculate sperm existing rate, malformed rate and sperm activity under 400 times light microscope. ③ The homogenizing mixture of one testis was made, and the mixture was centrifuged ,and the whole supernatant was taken to measure the activity of testicular superoxide dismutase （SOD）, glotathione-SH peroxidase （GSH-Px）, catalase （CAT）and the level of testicular malondialdehyde （MDA）. Testis constitution structure was observed by light mirror after haematine-eosin staining. ④ Condition of 100 sperm of seminiferous tubule was observed by light microscope. Different effects of cyclophosphamide and vitamin E were evaluated by anti-spermatogenesis effect integration （normal spermatogenesis： 0 points, 15% spermatogenesis obstacle of seminiferous tubule： 1 points, 15%-30%spermatogenesis obstacle of seminiferous tubule： 2 points, 30%-40% spermatogenesis obstacle of seminiferous tubule： 3 points, 40%-60% spermatogenesis obstacle of seminiferous tubule： 4 points, 60%-75% spermatogenesis obstacle of seminiferous tubule： 5 points, 75%-90% spermatogenesis obstacle of scminiferous tubule： 6 points, over 90% spermatogenesis obstacle of seminiferous tubule： 7 points, over 90% spermatogenesis obstacle of seminiferous tubule with a little spermatogenous cells： 8 points）. RESULTS： Twenty-four mice were involved in the analysis of results. ① As compared with the normal control group in the cyclophosphamide group, the plasma level of T, FSH,LH and the activity of testicular SOD,GSH-Px, CAT and sperm cout, motility, survival rate, were significantly decreased （P〈 0.05-0.01）. The MDA level and sperm abnormal forms rate were significantly higher than those in control group （P 〈 0.01）. Stratification of spermatogenic cell of testis constitution had small number, and lesion such as multinuclear giant cell and necrosis etc. was appeared. Anti-sperm effect was higher than that in control group [（6.96＋0.83 ）,0 point, （P 〈 0.01）]. ② As compared with the cyclophosphamide group in vitamin E treatment group, the plasma level of T,FSH,LH and the activity of testicular SOD, GSH-Px, CAT were significantly higher （P 〈 0.05-0.01）, and sperm teratism rate, and the content of MDA in testis constitution was lower than that in cyclophosphamide group （P 〈 0.01 ）. The testicular histological structure were significantly improved compared with cyclophosphamide group, while the anti-sperm effect was lower than that in cyclophosphamide group [（0.70±0.55 ）, （6.96±0.83）points ,P 〈 0.01]. CONCLUSION： Vitamin E has a signficantly protective effect on spermatogenic damage by cyclophosphamide. The action of vitamin E maybe have relationship with its scavenging free radical and anti-oxidation effect, as well as the acceleration of gonadotrophin releasing of pituitary anterior lobe etc.
出处
《中国临床康复》
CSCD
北大核心
2005年第27期74-76,共3页
Chinese Journal of Clinical Rehabilitation
