【摘要】：實(shí)驗(yàn)一Ouabain誘導(dǎo)大鼠弱精子癥模型建立的實(shí)驗(yàn)研究 目的為探討Ouabain引起弱精子癥的病理機(jī)制,建立SD大鼠腹腔注射Ouabain誘導(dǎo)的弱精子癥動(dòng)物模型。 方法健康性成熟SD雄性大鼠40只,隨機(jī)分為對(duì)照組10只(生理鹽水10mg/kg/d),低劑量實(shí)驗(yàn)組15只(Ouabain 12.5ug/kg/d),高劑量實(shí)驗(yàn)組15只(Ouabain 25ug/kg/d),連續(xù)腹腔注射30d,觀察各組大鼠附睪精子活力及活動(dòng)率的變化,同時(shí)觀察本實(shí)驗(yàn)劑量范圍的Ouabain對(duì)大鼠生殖系統(tǒng)及全身可能的毒性反應(yīng)。 結(jié)果與對(duì)照組相比,實(shí)驗(yàn)組大鼠精子數(shù)量無明顯變化,精子活力及活動(dòng)率顯著下降。高劑量及低劑量Ouabain組連續(xù)給藥30d后大鼠附睪精子活力分別為48.27%和44.73%,與對(duì)照組(73.33%)相比有顯著性差異(p0.01),且高、低劑量組間差異有顯著性(p0.01);高、低劑量組大鼠精子活動(dòng)率分別為52.26%和49.95%,與對(duì)照組(78.70%)相比有顯著性差異(p0.01),高、低劑量組間差異亦有顯著性(p0.01);本試驗(yàn)劑量范圍內(nèi),Ouabain對(duì)大鼠生殖器官及全身重要臟器未顯示明顯的毒性反應(yīng)。 結(jié)論Ouabain經(jīng)腹腔注射給藥30d能成功誘導(dǎo)SD大鼠弱精子癥模型的建立,且未見明顯的生殖系統(tǒng)和全身毒性反應(yīng)。 實(shí)驗(yàn)二小鼠睪丸間質(zhì)細(xì)胞分泌Ouabain的探索性研究 目的檢測(cè)體外培養(yǎng)的睪丸間質(zhì)細(xì)胞經(jīng)hCG、FSH干預(yù)后分泌Ouabain的功能,通過對(duì)小鼠睪丸間質(zhì)細(xì)胞功能的研究來探討其與Ouabain及弱精子癥之間的關(guān)系。 方法體外培養(yǎng)小鼠睪丸間質(zhì)細(xì)胞,用不同濃度的hCG(0U, 4U,8U,16U,32U,64U,100U,200U)和FSH(0,5,10,20,50,100mU/ml)對(duì)小鼠睪丸間質(zhì)細(xì)胞進(jìn)行干預(yù),收集上清,ELISA競(jìng)爭(zhēng)法檢測(cè)待測(cè)上清內(nèi)的Ouabain水平。 結(jié)果在hCG刺激下的體外培養(yǎng)的小鼠睪丸間質(zhì)細(xì)胞上清液中能夠檢測(cè)到Ouabain,在4U hCG作用下Ouabain水平為49.82pg/ml,在8U hCG作用下Ouabain水平增加為60.70 pg/ml,而在200U hCG作用下Ouabain水平達(dá)259.13 pg/ml;在一定濃度范圍FSH刺激作用下,體外培養(yǎng)小鼠睪丸間質(zhì)細(xì)胞上清液中未能檢測(cè)到Ouabain。 結(jié)論小鼠睪丸間質(zhì)細(xì)胞能夠合成Ouabain,且在一定劑量范圍內(nèi)分泌Ouabain的水平隨hCG刺激濃度的增高而升高,存在一定的量效關(guān)系;但在本實(shí)驗(yàn)劑量范圍FSH刺激下對(duì)小鼠睪丸間質(zhì)細(xì)胞未見影響。
[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.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R698;R-332

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