慢性应激条件下雄性大鼠生殖行为和睾丸细胞形态学的改变
发布时间：2019-06-19 22:13
【摘要】：目的:许多临床研究表明,慢性应激是引发人类生殖功能障碍的重要因素。本研究采用慢性轻度不可预见性应激与孤养相结合的方法建立慢性应激大鼠模型,探索慢性应激对大鼠生殖行为的影响,并从形态学角度检验睾丸生精细胞和间质细胞在慢性应激作用下的形态学改变状况。以此探讨慢性应激条件下,生殖行为和生殖功能改变的机制问题。 方法:16只雄性SD大鼠随机分为对照组和模型组(n=8),模型组采用慢性轻度不可预见性应激加孤养刺激大鼠,连续进行35天,构建模型,对照组正常饲养大鼠。整个实验过程中连续检测大鼠体重,结束后使用糖水偏爱实验和旷场实验检测大鼠的情绪状态,以确认造模成功。接近行为的检测使用Agmo提出的雌鼠偏爱的实验方法,用雌鼠偏爱分数和雌/雄区停留时间反映大鼠的接近行为。行为检测结束后牺牲大鼠,取睾丸组织石蜡包埋,HE染色后光学显微镜下观察睾丸间质和生精小管的形态,测量间质细胞面积、数量;测量生精小管直径、生精上皮厚度及一定区域内细胞数,并作统计分析。与此同时,树脂包埋,超薄切片后,在透射电子显微镜下观察对照组、模型组睾丸间质细胞和生精细胞的细胞结构和细胞器形态,并对比发现其中的差异。 结果:(1)慢性应激对情绪状态的影响:在慢性应激作用下从14天起模型组大鼠体重显著少于对照组,并持续至35天。造模结束后与对照组相比,模型组大鼠糖水偏爱分数减少(p0.01),旷场实验中行进距离、站立次数显著减少(p0.05,p0.01),中央格停留时间显著延长(p0.05),过中央格次数、粪便粒数显著增多(p0.01,p0.05)。显示动物模型制作成功。 (2)慢性应激对接近行为的影响:模型组自身对比,雌鼠处于发情期或发情间期对模型组雄鼠的雌鼠偏爱分数不影响;对雌/雄鼠区停留时间无影响。与对照组对比,雌鼠为发情间期时,模型组雌鼠偏爱分数显著减小(p0.001);模型组雄鼠雌鼠区停留时间显著减少(p0.01),雄鼠区停留时间显著增加(p0.01)。 (3)慢性应激对睾丸影响的光镜检测结果:与对照组相比,模型组大鼠生精小管管腔内的精子明显稀少,模型组生精小管短轴平均直径(p0.01)、生精上皮平均厚度(p0.01)和生精细胞数(p0.001)均小于对照组;睾丸间质面积(p0.01)、单个间质细胞面积(p0.01)睾丸间质细胞数(p0.01)均小于对照组,且统计结果有显著性意义。 (4)慢性应激对睾丸影响的电镜检测结果:与对照组正常细胞相比,模型组精原细胞细胞质与细胞核间出现空泡;精母细胞内质网扩张,细胞质出现空泡,线粒体肿胀;精子细胞内质网肿胀甚至形成空泡。精子鞭毛结构紊乱,外周致密纤维出现断裂和空泡。间质细胞细胞核出现深凹陷,染色质聚集,出现细胞凋亡现象。 结论:慢性应激环境可以导致雄性大鼠发生生殖行为抑制;并且引发睾丸间质细胞损伤和凋亡,以及生精细胞的受损。慢性应激导致大鼠生殖行为的减退可能与睾丸间质细胞受损,睾酮水平下降有关。
[Abstract]:Objective: Many clinical studies have shown that chronic stress is an important factor to induce human reproductive dysfunction. In this study, the model of chronic stress rats was established by the combination of chronic mild unpredictability and isolation, and the effects of chronic stress on the reproductive behavior of rats were explored, and the morphological changes of the spermatogenic cells and the interstitial cells under the effects of chronic stress were examined from the morphological angle. In this paper, the mechanism of the change of reproductive and reproductive function under the condition of chronic stress is discussed. Methods:16 male SD rats were randomly divided into control group and model group (n = 8). Rat body weight was detected continuously throughout the course of the experiment. After the end of the experiment, the experimental and open-field experiment was used to test the emotional state of the rats, so as to confirm the model. The test method of the female mouse preference proposed by the Agmo for the detection of the proximity behavior, using the female mouse preference score and the female/ male region residence time to reflect the proximity of the rat In order to sacrifice the rat after the end of the behavior detection, the testis tissue was embedded in paraffin, and the morphology of the testis and the seminiferous tubules was observed under the optical microscope after HE staining, the area and the number of the interstitial cells were measured, the diameter of the spermatogenic tube, the thickness of the spermatogenic epithelium and the cells in a certain region were measured. Count and make statistics In the meantime, the cell structure and organelle morphology of the testis and the spermatogenic cells in the control group and the model group were observed under the transmission electron microscope at the same time, and the difference was found in the model group. The results were as follows: (1) The effect of chronic stress on the emotional state: The weight of rats in the model group was significantly lower than that of the control group under the effect of chronic stress. Compared with the control group, the preference score of sugar water in the model group decreased (p0.01) and the number of standing times was significantly reduced (p0.05, p0.01), and the time of the central cell was significantly prolonged (p0.05), the number of cells in the central cell and the number of the grains per day (p0.01, p0). 05). Display animal model system to be successful. (2) The effect of chronic stress on the approach behavior: the model group itself vs. the female rats in the estrus or oestrus interval did not affect the female preference score of the male rats in the model group; the female/ male mouse area was stopped There was no effect on the time. In contrast to the control group, the preference score of the female rats in the model group was significantly decreased (p0.001), while the residence time of the female rats in the model group was significantly decreased (p0.01), and the residence time of the male rats was significantly increased (p (3) The results of light microscopy on the effects of chronic stress on the testis: compared with the control group, the sperm in the cavity of the model group was significantly less than that of the control group, and the average diameter of the short axis of the seminiferous tubule in the model group was less than that of the control group. (p0.01), the average thickness of the spermatogenic epithelium (p0.01) and the number of spermatogenic cells (p0.01) were all smaller than that of the control group, the interstitial area of the testis (p0.01), the area of the individual interstitial cells (p0.01) and the number of the testis interstitial cells (p0.01) were all smaller than that of the control group, and the statistical junction The effect of chronic stress on the testis was found to be significant. (4) The results of the electron microscopic examination of the effect of chronic stress on the testis: the vacuoles between the cytoplasm and the nucleus of the spermatogonia were observed in the model group compared with the normal cells in the control group, and the endoplasmic reticulum of the spermatocytes expanded and the cytoplasm was out. vacuolation The swelling of the net and even the formation of vacuoles. The structure of the flagellum of the sperm is disordered and the outer Breakage and vacuolation of the fibers. The nucleus of the interstitial cells has a deep depression, and the chromatin is concentrated. Conclusion: The chronic stress environment can lead to the inhibition of the reproductive behavior of the male rats. The death, and the damage to the spermatogenic cells. Chronic stress can lead to a decrease in the reproduction behavior of the rat and the interstitial cells of the testis.
【学位授予单位】：浙江理工大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363
本文编号：2502677
[Abstract]:Objective: Many clinical studies have shown that chronic stress is an important factor to induce human reproductive dysfunction. In this study, the model of chronic stress rats was established by the combination of chronic mild unpredictability and isolation, and the effects of chronic stress on the reproductive behavior of rats were explored, and the morphological changes of the spermatogenic cells and the interstitial cells under the effects of chronic stress were examined from the morphological angle. In this paper, the mechanism of the change of reproductive and reproductive function under the condition of chronic stress is discussed. Methods:16 male SD rats were randomly divided into control group and model group (n = 8). Rat body weight was detected continuously throughout the course of the experiment. After the end of the experiment, the experimental and open-field experiment was used to test the emotional state of the rats, so as to confirm the model. The test method of the female mouse preference proposed by the Agmo for the detection of the proximity behavior, using the female mouse preference score and the female/ male region residence time to reflect the proximity of the rat In order to sacrifice the rat after the end of the behavior detection, the testis tissue was embedded in paraffin, and the morphology of the testis and the seminiferous tubules was observed under the optical microscope after HE staining, the area and the number of the interstitial cells were measured, the diameter of the spermatogenic tube, the thickness of the spermatogenic epithelium and the cells in a certain region were measured. Count and make statistics In the meantime, the cell structure and organelle morphology of the testis and the spermatogenic cells in the control group and the model group were observed under the transmission electron microscope at the same time, and the difference was found in the model group. The results were as follows: (1) The effect of chronic stress on the emotional state: The weight of rats in the model group was significantly lower than that of the control group under the effect of chronic stress. Compared with the control group, the preference score of sugar water in the model group decreased (p0.01) and the number of standing times was significantly reduced (p0.05, p0.01), and the time of the central cell was significantly prolonged (p0.05), the number of cells in the central cell and the number of the grains per day (p0.01, p0). 05). Display animal model system to be successful. (2) The effect of chronic stress on the approach behavior: the model group itself vs. the female rats in the estrus or oestrus interval did not affect the female preference score of the male rats in the model group; the female/ male mouse area was stopped There was no effect on the time. In contrast to the control group, the preference score of the female rats in the model group was significantly decreased (p0.001), while the residence time of the female rats in the model group was significantly decreased (p0.01), and the residence time of the male rats was significantly increased (p (3) The results of light microscopy on the effects of chronic stress on the testis: compared with the control group, the sperm in the cavity of the model group was significantly less than that of the control group, and the average diameter of the short axis of the seminiferous tubule in the model group was less than that of the control group. (p0.01), the average thickness of the spermatogenic epithelium (p0.01) and the number of spermatogenic cells (p0.01) were all smaller than that of the control group, the interstitial area of the testis (p0.01), the area of the individual interstitial cells (p0.01) and the number of the testis interstitial cells (p0.01) were all smaller than that of the control group, and the statistical junction The effect of chronic stress on the testis was found to be significant. (4) The results of the electron microscopic examination of the effect of chronic stress on the testis: the vacuoles between the cytoplasm and the nucleus of the spermatogonia were observed in the model group compared with the normal cells in the control group, and the endoplasmic reticulum of the spermatocytes expanded and the cytoplasm was out. vacuolation The swelling of the net and even the formation of vacuoles. The structure of the flagellum of the sperm is disordered and the outer Breakage and vacuolation of the fibers. The nucleus of the interstitial cells has a deep depression, and the chromatin is concentrated. Conclusion: The chronic stress environment can lead to the inhibition of the reproductive behavior of the male rats. The death, and the damage to the spermatogenic cells. Chronic stress can lead to a decrease in the reproduction behavior of the rat and the interstitial cells of the testis.
【学位授予单位】：浙江理工大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363
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