【作者】 王根辈；

【导师】 范光丽； 徐永平；

【作者基本信息】 西北农林科技大学 ， 神经生物学， 2008， 硕士

【摘要】 动情周期是雌性动物重要的生理周期之一,而作为孕育生命场所的子宫,其周期性变化则随着神经、免疫、内分泌系统的调节而发生相应的变化,三者既相互协同又相互制约。目前国内外对NSE、S100蛋白的研究多集中在通过测定外周血中的含量间接用于神经系统疾病的诊断、判断疾病严重程度、估计预后和指导治疗等方面,侧重于将其作为神经元损伤的标志物,忽略了NSE、S100蛋白和突触素本身还具有独特的生理作用,本试验采用免疫组织化学超敏感SP法和图像分析法对动情周期大鼠子宫中NSE、突触素和S100蛋白的分布和变化规律进行了研究,以期从子宫能量代谢、神经支配的突触终末和参与调节子宫功能作用的酸性钙离子结合蛋白三方面进行动情周期方面的研究,结果如下:1. NSE在大鼠动情周期子宫中的表达定位及变化规律:子宫各层均有不同程度NSE免疫阳性产物的分布,并随动情周期的变化呈现如下规律:子宫内膜中,动情期着色最深,动情后期、动情间期、动情前期着色依次减弱;子宫肌层和外膜中,动情期着色最深,动情后期着色最浅,动情间期表达量明显回升,动情前期比动情期着色稍浅,提示NSE的表达可能与雌激素表达呈正相关,是影响动情周期规律性变化的因子之一。2.突触素在大鼠动情周期子宫中的表达定位及变化规律:子宫各层均有不同程度突触素免疫阳性产物分布,并随动情周期的变化呈现如下规律:各期子宫内膜着色强弱变化不大,但动情前期着色最深,动情后期次之,动情期、动情间期着色最浅;子宫肌层和外膜中,动情前期着色最深,动情期着色减弱,动情后期着色最浅,动情间期表达量明显回升。揭示突触素在大鼠动情周期子宫中的表达分布变化具有规律性,可能与雌激素、孕激素具有一定的相关性。3. S100蛋白在大鼠动情周期子宫中的表达定位及变化规律:子宫各层均有不同程度S100蛋白免疫阳性产物的分布,并随动情周期的发生而表现如下规律:各期子宫内膜着色深浅有一定差别,动情前期着色最弱;动情期着色最深,表达升高;动情后期降低,但到动情间期S100蛋白表达重新升高,表达呈双峰趋势;子宫肌层和外膜中,各期着色差异不显著,但具有如下变化趋势:动情前期着色较浅,动情期着色最深,动情后期着色又变浅,但动情间期着色明显升高,表达亦有双峰趋势。提示S100蛋白的表达可能受卵巢类固醇激素的调控,在动情周期子宫中发挥重要生理作用,作为一类酸性钙结合蛋白,推测其可能参与调节子宫内局部酸碱度的变化,为精子获能提供适宜环境。更多还原

【Abstract】 As we know, uterus is the place of recreation, while as one of the most important physiological cycle, estrous cycle changes with neuro-immuno-endocrine regulation whichcan be enhanced or inhibited by each other. At present, most internal or external researches on NSE and S100 are focus on the diagnosis of neurosystem diseases, the level of diseases and the monitor of prognosis and instruction for treatment through determining the their content in the peripheral blood, taking them as the markers of neuron damage. These research fields ignored the specific per se physiological actions of NSE, S100 and synaptophysin. Our research utilize immunohistochemical SP method and the image analysis to analyze the distribution and variation of NSE, S100 protein and synaptophysin in the uterus of rat during its estrus which is destined to further understand estrous cycle from three aspects of energy metabolism, neuron allocation of synaptic terminus and acidic calcium binding protein which is involved in regulating the uterus functions. The results are as follows:1. The expression and variation of NSE in uterus in rat estrous cycle: The results shows that immunoactive products of NSE are present in each layer of uterus, though the intensity differs. And the distribution of the products changes with variation of estrus cycle which was found to follow certain rules: In endometrium, the density of coloration is strongest in the estrum, and then becomes weaker gradually in the metoestrum, anoestrum and proestrum. In muscular layer and adventitia, the density of coloration is strongest in the estrum and weakest in metoestrum, then the significant increase of the products was found in the anoestrum, and the density of the products was a little bit weaker in the proestrum than in the estrum, which imply that expression of NSE in the myometrium is positively related to estrogen, by which we speculate that NSE, as one factor affecting regularity variation of estrous cycle, may has the function in promoting matrix cell’s proliferation and decidualization, mucilage formation and secretion, as well as mating process.2. The expression and variation of Synaptophysin in uterus in rat estrous cycle: The results indicate that synaptophysin-IR products were present in each layer of uterus, though the intensity differs, and the distribution of the products in uterus changes with variation of estrus cycle which was found to follow certain rules: There is no significant difference of the expression of synaptophysin in the endometrium during every period. But the staining was strongest in the preoestrus, then it is weaker in metoestrus, and estrum and diestrus is lightest staining. In muscular layer and adventitia, the coloration was densest in the preoestrus, the significant increase of the products was found in the anoestrum, and the density of the products was a little bit weaker in the estrus than that in the metoestrus. This indicates that the distribution of Synaptophysin in uterus in rat estrus cycle has some regularity, and is related to estrogen and the progestogen. It is extrapolated that the estrogen may promote the Synaptophysin expression, the promotion of synapse generation, and increase the neuroncontact, on which they are probably opposite regulated by progestogen.3. The expression and variation of S100 protein in uterus in rat estrous cycle: uterine layers present different levels of immune S100 positive product distribution in estrous cycle and the expression is as follows: The staining intensity of immune-S100 positive cells is different in endometrial of different stages. The staining is weakest in proestrum,densest in estrum and decreased in postestrum. The expression of S100 is re-increased in metestrus which presents the bimodal distribution. There is no much difference in the expression of S100 protein in muscular layer in different periods, but there are some rules to follow: the expression of S100 protein in proestrus is slight, then it is strongly expressed in estrus, while it is decreased in metoestrus, which indicate that the expression of S100 which functions in uterus estrus cycle, is regulated by ovary steroid hormones. This implies that S100, as a kind of acidic calcium binding protein, may be involved in regulating pH values of uterus in order to give rise to proper environment for capacitation.更多还原