己烯雌酚（DES）對小鼠睪丸促性腺激素及其受體影響的實驗性研究
發(fā)布時間：2019-01-06 14:13
【摘要】：目的：研究表明人類乃至整個動物界的生殖發(fā)育都正受到日益嚴重的環(huán)境雌激素的影響，但環(huán)境雌激素對生殖系統(tǒng)發(fā)育影響的機理卻很不清楚。睪丸是雄性生殖系統(tǒng)的核心，促性腺激素(黃體生成素LH和卵泡刺激素FSH)是生殖系統(tǒng)發(fā)育過程中的重要激素，它們上連大腦下連睪丸并通過與睪丸上的相應受體結合而產生作用。近來，更有研究表明環(huán)境雌激素在低劑量和高劑量時對不同組織所產生的效應可以不同。環(huán)境雌激素對睪丸發(fā)育的影響機制目前非常不清楚。本實驗擬采用公認的環(huán)境雌激素己烯雌酚(DES)作用于孕期母鼠，然后通過檢測仔鼠發(fā)育過程中睪丸組織中的促性腺激素及其受體的變化情況以探討不同劑量外源性雌激素對睪丸乃至整個生殖系統(tǒng)發(fā)育的影響機理。 材料和方法：8～10周齡雌性昆明小鼠200只隨機分成8組(每組25只)：正常組、對照組、實驗組1、實驗組2、實驗組3、實驗組4、實驗組5、實驗組6。實驗組1-6分別給予DES 0.1、0.5、10.0、25.0、50.0及100.0μg．kg~(-1)．d~(-1)。DES以DMSO和生理鹽水為溶劑。對照組僅給予DMSO和生理鹽水；正常組不給任何藥物。雌雄小鼠按常規(guī)合籠交配，以出現(xiàn)陰栓作為受孕標志。發(fā)現(xiàn)陰栓當天定為妊娠第0天，記為GD0(gestation day，GD)。在GD9～GD17上午8～9時皮下注射給藥，每組所用的DES均溶于等量的DMSO及生理鹽水中，以保證每次每只小鼠每千克體重所給予的DMSO和生理鹽水相同。新生小鼠出生后，以小鼠出生第一天記為0，分別于0，5，15，25，35，45，60天處死6只雄性小鼠并取其睪丸：剖開部分睪丸固有鞘膜后固定于10％福爾馬林固定液中，，24小時后給予梯度酒精脫水，石蠟包埋。作4μm厚冠狀位連續(xù)切片進行常規(guī)的HE組織學染色觀察和特殊的免疫組織化學研究。應用光鏡觀察睪丸結構的變化，免疫組織化學方法分別檢測各組睪丸組織中的黃體生成素(LH)、黃體生成素受體(LHR)、卵泡刺激素(FSH)和卵泡刺激素受體(FSHR)，測定睪丸組織中各指標的陽性灰度值，以(?)±s表示，并采用方差分析和相關分析進行統(tǒng)計學分析。 結果：1．光鏡觀察可見正常組、對照組及實驗組的各級生精細胞、間質細胞和支持細胞形態(tài)上無明顯差異，但在100.0和50.0μg．kg~(-1)．d~(-1)組出現(xiàn)：①睪丸生精細胞層減少、稀疏，嚴重的廣泛性減少至細胞僅剩1～2層，睪丸生精細胞脫落至
[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 (?) 鹵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.
【學位授予單位】：汕頭大學
【學位級別】：碩士
【學位授予年份】：2006
【分類號】：R363
本文編號：2402899
[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 (?) 鹵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.
【學位授予單位】：汕頭大學
【學位級別】：碩士
【學位授予年份】：2006
【分類號】：R363
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