己烯雌酚（DES）对小鼠睾丸促性腺激素及其受体影响的实验性研究

发布时间：2019-01-06 14:13

【摘要】：目的：研究表明人类乃至整个动物界的生殖发育都正受到日益严重的环境雌激素的影响，但环境雌激素对生殖系统发育影响的机理却很不清楚。睾丸是雄性生殖系统的核心，促性腺激素(黄体生成素LH和卵泡刺激素FSH)是生殖系统发育过程中的重要激素，它们上连大脑下连睾丸并通过与睾丸上的相应受体结合而产生作用。近来，更有研究表明环境雌激素在低剂量和高剂量时对不同组织所产生的效应可以不同。环境雌激素对睾丸发育的影响机制目前非常不清楚。本实验拟采用公认的环境雌激素己烯雌酚(DES)作用于孕期母鼠，然后通过检测仔鼠发育过程中睾丸组织中的促性腺激素及其受体的变化情况以探讨不同剂量外源性雌激素对睾丸乃至整个生殖系统发育的影响机理。材料和方法：8～10周龄雌性昆明小鼠200只随机分成8组(每组25只)：正常组、对照组、实验组1、实验组2、实验组3、实验组4、实验组5、实验组6。实验组1-6分别给予DES 0.1、0.5、10.0、25.0、50.0及100.0μg．kg~(-1)．d~(-1)。DES以DMSO和生理盐水为溶剂。对照组仅给予DMSO和生理盐水；正常组不给任何药物。雌雄小鼠按常规合笼交配，以出现阴栓作为受孕标志。发现阴栓当天定为妊娠第0天，记为GD0(gestation day，GD)。在GD9～GD17上午8～9时皮下注射给药，每组所用的DES均溶于等量的DMSO及生理盐水中，以保证每次每只小鼠每千克体重所给予的DMSO和生理盐水相同。新生小鼠出生后，以小鼠出生第一天记为0，分别于0，5，15，25，35，45，60天处死6只雄性小鼠并取其睾丸：剖开部分睾丸固有鞘膜后固定于10％福尔马林固定液中，，24小时后给予梯度酒精脱水，石蜡包埋。作4μm厚冠状位连续切片进行常规的HE组织学染色观察和特殊的免疫组织化学研究。应用光镜观察睾丸结构的变化，免疫组织化学方法分别检测各组睾丸组织中的黄体生成素(LH)、黄体生成素受体(LHR)、卵泡刺激素(FSH)和卵泡刺激素受体(FSHR)，测定睾丸组织中各指标的阳性灰度值，以(?)±s表示，并采用方差分析和相关分析进行统计学分析。结果：1．光镜观察可见正常组、对照组及实验组的各级生精细胞、间质细胞和支持细胞形态上无明显差异，但在100.0和50.0μg．kg~(-1)．d~(-1)组出现：①睾丸生精细胞层减少、稀疏，严重的广泛性减少至细胞仅剩1～2层，睾丸生精细胞脱落至
[Abstract]:Objective: the study shows that the reproductive development of human beings and even the whole animal kingdom is affected by the increasingly serious environmental estrogens, but the mechanism of the effects of environmental estrogens on the development of the reproductive system is not clear. Testis are the core of male reproductive system. Gonadotropin (luteinizing hormone LH and follicle stimulating hormone FSH) are important hormones in the development of reproductive system. They connect to the testicles below the brain and act by binding to the corresponding receptors on the testicles. More recent studies have shown that environmental estrogens have different effects on different tissues at low and high doses. The effect of environmental estrogen on testicular development is unclear. In this study, the effect of diethylstilbestrol (DES), a recognized environmental estrogen, on pregnant rats was studied. Then the effects of different doses of exogenous estrogen on the development of testis and the whole reproductive system were studied by detecting the changes of gonadotropin and its receptors in testis during the development of newborn mice. Materials and methods: 200 10-week-old female Kunming mice were randomly divided into 8 groups (25 in each group): normal group, control group, experimental group 1, experimental group 2, experimental group 3, experimental group 4, experimental group 5, experimental group 6. Experimental group 1-6 was treated with DES 0.1g 路kg ~ (-1) and 100.0 渭 g 路kg ~ (-1) respectively. DMSO and normal saline were used as solvent. The control group was given only DMSO and normal saline, while the normal group was not given any drugs. The female and male mice were mated in a cage, and the vaginal suppository was used as the sign of conception. The day of discovery of Yin suppository was determined as day 0 of pregnancy, which was recorded as GD0 (gestation day,GD). When GD9~GD17 was given subcutaneously from 8 to 9 am, the DES of each group was dissolved in the same amount of DMSO and saline, so as to ensure that each mouse was given the same amount of DMSO and saline per kilogram of body weight. On the first day of birth, six male mice were killed on the first day of birth, and 6 male mice were killed and their testis were taken. The testicular sheath was cut open and fixed in 10% formalin fixative solution. After 24 hours, gradient alcohol dehydration and paraffin embedding were given. 4 渭 m thick coronal serial sections were used for routine HE staining and special immunohistochemical studies. The changes of testis structure were observed under light microscope. The luteinizing hormone (LH),) luteinizing hormone receptor (LHR),) follicle stimulating hormone (FSH) and follicle stimulating hormone receptor (FSHR),) in testis were detected by immunohistochemical method. The positive grayscale values of each index in testicular tissue were measured, expressed as (?) 卤s, and analyzed statistically by ANOVA and correlation analysis. Results: 1. There was no significant difference in morphology of spermatogenic cells, interstitial cells and Sertoli cells between normal group, control group and experimental group under light microscope. However, in the 100.0 and 50.0 渭 g 路kg ~ (-1) 路d ~ (-1) groups: (1) testicular spermatogenic cell layer decreased, sparse, severe extensiveness reduced to only 1 ~ 2 layers of cells, testicular spermatogenic cells fell off.
【学位授予单位】：汕头大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R363

【参考文献】