Ouabain诱导大鼠弱精子症模型建立及睾丸间质细胞分泌Ouabain的探索性研究
发布时间：2018-11-20 20:01
【摘要】：实验一Ouabain诱导大鼠弱精子症模型建立的实验研究 目的为探讨Ouabain引起弱精子症的病理机制,建立SD大鼠腹腔注射Ouabain诱导的弱精子症动物模型。 方法健康性成熟SD雄性大鼠40只,随机分为对照组10只(生理盐水10mg/kg/d),低剂量实验组15只(Ouabain 12.5ug/kg/d),高剂量实验组15只(Ouabain 25ug/kg/d),连续腹腔注射30d,观察各组大鼠附睾精子活力及活动率的变化,同时观察本实验剂量范围的Ouabain对大鼠生殖系统及全身可能的毒性反应。 结果与对照组相比,实验组大鼠精子数量无明显变化,精子活力及活动率显著下降。高剂量及低剂量Ouabain组连续给药30d后大鼠附睾精子活力分别为48.27%和44.73%,与对照组(73.33%)相比有显著性差异(p0.01),且高、低剂量组间差异有显著性(p0.01);高、低剂量组大鼠精子活动率分别为52.26%和49.95%,与对照组(78.70%)相比有显著性差异(p0.01),高、低剂量组间差异亦有显著性(p0.01);本试验剂量范围内,Ouabain对大鼠生殖器官及全身重要脏器未显示明显的毒性反应。 结论Ouabain经腹腔注射给药30d能成功诱导SD大鼠弱精子症模型的建立,且未见明显的生殖系统和全身毒性反应。 实验二小鼠睾丸间质细胞分泌Ouabain的探索性研究 目的检测体外培养的睾丸间质细胞经hCG、FSH干预后分泌Ouabain的功能,通过对小鼠睾丸间质细胞功能的研究来探讨其与Ouabain及弱精子症之间的关系。 方法体外培养小鼠睾丸间质细胞,用不同浓度的hCG(0U, 4U,8U,16U,32U,64U,100U,200U)和FSH(0,5,10,20,50,100mU/ml)对小鼠睾丸间质细胞进行干预,收集上清,ELISA竞争法检测待测上清内的Ouabain水平。 结果在hCG刺激下的体外培养的小鼠睾丸间质细胞上清液中能够检测到Ouabain,在4U hCG作用下Ouabain水平为49.82pg/ml,在8U hCG作用下Ouabain水平增加为60.70 pg/ml,而在200U hCG作用下Ouabain水平达259.13 pg/ml;在一定浓度范围FSH刺激作用下,体外培养小鼠睾丸间质细胞上清液中未能检测到Ouabain。 结论小鼠睾丸间质细胞能够合成Ouabain,且在一定剂量范围内分泌Ouabain的水平随hCG刺激浓度的增高而升高,存在一定的量效关系;但在本实验剂量范围FSH刺激下对小鼠睾丸间质细胞未见影响。
[Abstract]:Experiment 1 the establishment of rat model of asthenospermia induced by Ouabain objective to investigate the pathological mechanism of asthenospermia induced by Ouabain and to establish an animal model of asthenospermia induced by intraperitoneal injection of Ouabain into SD rats. Methods Forty healthy mature male SD rats were randomly divided into three groups: control group (n = 10), low dose group (Ouabain 12.5ug/kg/d) and high dose group (Ouabain 25ug/kg/d). The changes of sperm motility and motility of epididymal sperm were observed after 30 days of continuous intraperitoneal injection. The possible toxicity of Ouabain in the range of dose to the reproductive system and the whole body of rats was also observed. Results compared with the control group, the number of spermatozoa in the experimental group did not change significantly, but the sperm motility and motility decreased significantly. The sperm motility of the epididymal sperm was 48.27% and 44.73% respectively in the high dose and low dose Ouabain groups after 30 days of continuous administration, which was significantly higher than that in the control group (73.33%) (p0.01). There was significant difference between the low dose groups (p0.01). The sperm motility rates in the high and low dose groups were 52.26% and 49.95% respectively, which were significantly higher than those in the control group (78.70%) (p0.01). In the range of dose, Ouabain showed no obvious toxicity to the reproductive organs and important organs of the whole body in rats. Conclusion the model of asthenospermia in SD rats can be successfully induced by intraperitoneal injection of Ouabain for 30 days, and there is no obvious reproductive system or systemic toxicity. An exploratory study on the secretion of Ouabain by the Leydig cells of the testes of the second Rat objective to investigate the function of Ouabain secreted by cultured interstitial cells of testis after hCG,FSH intervention in vitro. The relationship between interstitial cell function of mouse testis and Ouabain and asthenospermia was studied. Methods Leydig cells of mouse testis were cultured in vitro. The interstitial cells of mouse testis were treated with different concentrations of hCG (0U, 4U 8U 16U 16U) and FSH (2050mU / ml), and the supernatants were collected. ELISA competition law was used to detect the level of Ouabain in the supernatant. Results in the supernatant of cultured mouse testicular stromal cells stimulated by hCG, the Ouabain level of Ouabain, was 49.82 PG / ml at 4U hCG, and the Ouabain level increased to 60.70 pg/ml, under 8U hCG. The level of Ouabain reached 259.13 pg/ml; under 200U hCG. Ouabain. was not detected in the supernatant of mouse testicular stromal cells cultured in vitro under the stimulation of certain concentration range of FSH. Conclusion the interstitial cells of mouse testis can synthesize Ouabain, and the level of endocrine Ouabain increases with the increase of the concentration of hCG in a certain dose range, which has a dose-effect relationship. However, there was no effect on the interstitial cells of mouse testis stimulated by FSH in the dose range of this experiment.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R698;R-332
本文编号：2345850
[Abstract]:Experiment 1 the establishment of rat model of asthenospermia induced by Ouabain objective to investigate the pathological mechanism of asthenospermia induced by Ouabain and to establish an animal model of asthenospermia induced by intraperitoneal injection of Ouabain into SD rats. Methods Forty healthy mature male SD rats were randomly divided into three groups: control group (n = 10), low dose group (Ouabain 12.5ug/kg/d) and high dose group (Ouabain 25ug/kg/d). The changes of sperm motility and motility of epididymal sperm were observed after 30 days of continuous intraperitoneal injection. The possible toxicity of Ouabain in the range of dose to the reproductive system and the whole body of rats was also observed. Results compared with the control group, the number of spermatozoa in the experimental group did not change significantly, but the sperm motility and motility decreased significantly. The sperm motility of the epididymal sperm was 48.27% and 44.73% respectively in the high dose and low dose Ouabain groups after 30 days of continuous administration, which was significantly higher than that in the control group (73.33%) (p0.01). There was significant difference between the low dose groups (p0.01). The sperm motility rates in the high and low dose groups were 52.26% and 49.95% respectively, which were significantly higher than those in the control group (78.70%) (p0.01). In the range of dose, Ouabain showed no obvious toxicity to the reproductive organs and important organs of the whole body in rats. Conclusion the model of asthenospermia in SD rats can be successfully induced by intraperitoneal injection of Ouabain for 30 days, and there is no obvious reproductive system or systemic toxicity. An exploratory study on the secretion of Ouabain by the Leydig cells of the testes of the second Rat objective to investigate the function of Ouabain secreted by cultured interstitial cells of testis after hCG,FSH intervention in vitro. The relationship between interstitial cell function of mouse testis and Ouabain and asthenospermia was studied. Methods Leydig cells of mouse testis were cultured in vitro. The interstitial cells of mouse testis were treated with different concentrations of hCG (0U, 4U 8U 16U 16U) and FSH (2050mU / ml), and the supernatants were collected. ELISA competition law was used to detect the level of Ouabain in the supernatant. Results in the supernatant of cultured mouse testicular stromal cells stimulated by hCG, the Ouabain level of Ouabain, was 49.82 PG / ml at 4U hCG, and the Ouabain level increased to 60.70 pg/ml, under 8U hCG. The level of Ouabain reached 259.13 pg/ml; under 200U hCG. Ouabain. was not detected in the supernatant of mouse testicular stromal cells cultured in vitro under the stimulation of certain concentration range of FSH. Conclusion the interstitial cells of mouse testis can synthesize Ouabain, and the level of endocrine Ouabain increases with the increase of the concentration of hCG in a certain dose range, which has a dose-effect relationship. However, there was no effect on the interstitial cells of mouse testis stimulated by FSH in the dose range of this experiment.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R698;R-332
【参考文献】
相关期刊论文 前6条
1 杨小红,张伟程;雷公藤制剂的应用及其副作用[J];中医药临床杂志;2004年05期
2 石其贤,张寅恭,袁玉英;棉酚抗生育作用的研究 Ⅱ.对雄性大鼠的抗生育作用[J];动物学报;1981年01期
3 孙晓梅;陈瑜;李春花;彭辉;林善良;代解杰;;食蟹猴少精弱精动物模型建立的研究[J];实验动物科学与管理;2006年02期
4 吕卓人;;内源性哇巴因的研究与展望[J];西安交通大学学报(医学版);2005年06期
5 刘梅颜,吕卓人,王颢;哇巴因、地高辛与高血压发病关系的研究[J];西安交通大学学报(医学版);2003年03期
6 葛蘅,吕卓人,于汶,张汉伟;哇巴因浓度检测方法与评估[J];中国实验诊断学;1999年03期
，本文编号：2345850
本文链接：https://www.wllwen.com/yixuelunwen/shiyanyixue/2345850.html
最近更新
教材专著
