【作者】 熊若虹；

【导师】 羊惠君；

【作者基本信息】 四川大学 ， 人体解剖与组织胚胎学， 2007， 博士

【摘要】 【目的】以建立雌性SD大鼠过量运动致运动性月经失调(Athletic menstrual cycle irregularities.AMI)模型为基础，研究运动疲劳对雌性生殖系统及骨形态和功能的影响，并初步探讨其机制。【材料与方法】1．模型建立：采用健康、均有正常动情周期的3月龄雌性SD大鼠140只，随机分为空白组和造模组，造模组100只，对照组40只。空白组大鼠不作任何干预，自然喂养至实验结束；造模组每天按设定的大负荷运动方案进行力竭游泳运动训练，每周训练6天，休息1天。两组大鼠每天上午8∶00～9∶00进行一次阴道脱落细胞检查，以动态观测大鼠阴道脱落细胞的变化，确定大鼠动情周期并计算出角化细胞所占的比例。2．造模成功后处死动物，无菌取其血液及卵巢、阴道中1／3、子宫、大脑半球组织进行一般形态学观察测量，并制作组织切片，观察其在显微、超微结构及受体表达等方面的改变。3．取两组新鲜卵巢组织，按Qbio-gene公司提供操作程序，平行操作，建立两组差异基因表达谱，并对其作初步比较分析。4．实验第22天用双能X线骨密度吸收仪测定两组大鼠全身骨密度。处死大鼠后，取大鼠右股骨上端2cm和腰4-6椎骨，剔净后测骨密度。对所得数据进行统计学处理。【结果】1．发现了长期力竭运动可使大鼠卵巢卵泡的发育、成熟、排卵、黄体形成等功能发生抑制，使颗粒细胞、卵泡膜细胞、黄体细胞等超微结构发生改变，细胞功能包括性激素的分泌功能受到抑制，性激素水平下降；长期力竭运动还对卵巢颗粒细胞、黄体细胞ER、PR的表达具有抑制作用。2．当大量研究都提示运动有利于促进骨骼生长、发育、改善骨代谢和提高骨密度，并可有效的预防和治疗骨质疏松症时，本研究发现长期大负荷运动训练可能对骨骼的健康产生负面影响，尤其是女性。3．RFDD-PCR结果显示：两种组织共获得了有意义的条带3512条，其中模型组卵巢组织1802条，对照组卵巢组织1710条，有354个基因片段为模型组卵巢所独有，336个片段在正常对照组中的特异显示。通过对差异表达谱中泛素-蛋白酶体系统进行生物信息学分析，发现3个蛋白酶体亚单位基因(PSs)、3个泛素特异蛋白酶基因(USPs)及泛素蛋白连接酶E3A基因(UBE3A)在模型组卵巢组织中过表达，提示运动疲劳所致生殖系统损伤过程中泛素-蛋白酶体系统活动明显加强。【结论】运动性疲劳运动可使大鼠生殖系统，尤其是卵巢等细胞功能包括性激素的分泌功能受到抑制，细胞超微结构发生改变，血清雌激素水平、睾酮水平下降，羟脯氨酸水平升高，阴道pH值增高、阴道脱落角化细胞大量减少等，提示可导致生殖系统和骨质的损伤。损伤涉及到广泛的基因表达变化，其中泛素-蛋白酶体系统相关基因活动明显加强。更多还原

【Abstract】 Objective: To build female SD rats AMI model due to over-loading exercise for the study of AMI in female athletes, to observe morphological and functional changes of reproductive system and skeleton under sports fatigue and explore its mechanism. Materiel and Method: 1. animal model: 140 healthy female SD rats, 3 month old, with normal menstrual cycle, were selected and divided into experimental group and control group randomly. No measure was taken to control group, while rats of experimental group were trained using over-loading swimming for 6 days per week, then after 1 day’s break, another 6 days’ training again until 22 days later. Detection of desquamated keratinocyte index of vagina was done at AM 8: 00～9: 00 every day for observe any possible changes on rats’ menstrual cycle 2. When AMI model was founded successfully 22 days later, rats of 2 groups were killed, samples of blood, tissues of ovary gland, uterus, middle 1/3 of vagina and cerebral hemisphere were collected for morphological observation and measurement. Tissue slides were made for observation IMH detection and microscope and electric-microscope observations. 3. 2 samples of ovary gland from 2 groups respectively, gene differential expression profiles were constructed on the same pace according to the protocol from Qbio-gene Company. Bioinformatics analysis was used to compare altered genes between 2 profiles. 4. on the 22^nd days of experiment, bone mineral density of rats from 2 groups were measured using X rays, upper part of right femur and 4^th to 6^th lumber vertebrae were taken to check bone mineral density again. Results: 1. almost all the functional process of reproductive system, especially ovary gland, were interfered by over-loading exercise. Cellular structures changes were observed under microscope. Levels of E2, P, T, GH, C, SOD, etc. decreased in experimental group compared with control group （p<0.05）. 2. measurement of mineral density of bones also showed that a decreasing tendency in experimental group. 3. Totally 3512 significant bands were displayed including 1802 of experimental ovary and 1710 in control tissue using RFDD-PCR. 354 bands were separated only in experimental group and 336 bands only in control group. Trough bioinformatics analysis on the differential expression profiles, over-expression members of UPS system were detected, including 3 PSs, 3 UPSs and UBE3A. Conclusion: sports fatigue can depress cellular function of rats’ reproductive system especially ovary gland including secretion of sex hormone, changes of cellular structure, level of esterone, PH value in vagina etc., showed the injury of reproductive system and bones. The mechanism of these changes may result from extensive genes differentially expression, among them, the activation of UPS system should be put emphasis on.更多还原