【作者】 姜忠玲；

【导师】 王林安；

【作者基本信息】 东北农业大学 ， 临床兽医学， 2006， 博士

【摘要】 以昆明雄性小白鼠为受试动物,分别用近似阴囊温度(33℃)、腹温(37℃)和超高温(42℃)三个温度组,实施1h/d的应激处理,以建立持续的全身热应激动物模型。以小鼠精子发生过程各阶段所需时间确定热应激持续的时间,每个温度组又分别设四个不同热应激持续时间(8d、13d、21d和35d)。在热应激处理的过程中记录小鼠应激前后体重和肛温的变化。分别在热应激持续到8d、13d、21d和35d时,应用形态学观察法,研究受试鼠睾丸和附睾重量及指数、组织结构、细胞凋亡和附睾精子形态学的变化;应用免疫荧光细胞化学和蛋白质印迹分析的方法,研究睾丸和附睾组织中热休克蛋白70(HSP70)的表达;应用比色法检测睾丸和附睾组织内活性氧(ROS)和丙二醛(MDA)、抗氧化物酶(SOD、GSH-Px、CAT)的活力或浓度的变化;应用交配试验,检测雄鼠受精能力及其致孕雌鼠的产仔数的变化,用以揭示热应激对雄鼠生殖机能的影响。试验结果如下:1.热应激前后的鼠体重和肛温变化体重的变化:受热应激前和后,鼠体重减轻,并且是热应激后的体重低于热应激前的体重。而受热应激前和后的鼠体重的差值变化是热应激温度越高其越大,热应激次数越多其越小。肛温的变化:受热应激后,鼠肛温上升,并且是热应激温度越高,上升的幅度越明显,但随热应激次数的增加变化不显著;而每次受热应激前,鼠肛温恢复到对照组的水平。2.热应激鼠体重、睾丸和附睾重量及其重量指数的变化受热应激后,与对照组相比,鼠的体重、睾丸和附睾重下降,并且热应激温度越高越明显。除37℃组和33℃组分别应激8d和13d外的各组,应激后的鼠体重均显著(P<0.05)低于对照组;除33℃组分别应激13d和21d、37℃组应激13d外的各组,应激后的鼠睾丸重量均显著(P<0.05)低于对照组;除33℃组和37℃组应激8d外的各组,应激后的鼠附睾重量均显著(P<0.05)低于对照组。除42℃应激后的鼠睾丸重量指数显著(P<0.05)小于对照组外,其它各组的鼠睾丸重量指数与对照组间均无显著差异(P>0.05)。而受热应激后,各应激组的鼠附睾重量指数与对照组间均无显著差异(P>0.05)。3.热应激鼠附睾尾精子的形态学变化受热应激后,鼠附睾尾精子密度和精子顶体完整率呈下降趋势,并低于对照组。除33℃组分别应激8d、13d后的精子密度与对照组间无显著差异(P>0.05)外,其它各组均显著(P<0.05)低于对照组。除33℃应激后的精子顶体完整率与对照组间无显著差异(P>0.05)外,其它各组均显著(P<0.05)低于对照组。受热应激后,鼠附睾尾精子畸形率呈现升高趋势,并高于对照组。除37℃组应激8d、33℃组分别应激8d、13d、21d后的精子畸形率与对照组无显著差异(P>0.05)外,其它各组均显著(P<0.05)高于对照组。4.热应激鼠睾丸和附睾组织病理学变化及细胞凋亡更多还原

【Abstract】 Male Kunming mice were exposed to heat stress within three groups at the temperature same as scrotum (33℃), abdomen (37℃)and severe high temperature (42℃), and the sustaining systemic heat stressed animal model were established by this way. Each group (8d, 13d, 21d and 35d) were divided into four groups with different duration of heat stress according to the phases of the process of spermatogenesis. The changes in the weight and rectal temperature were registered during the process of heat stress. The changes in the weight, exponential, structure of the tissue, cell apoptosis in testis and epididymis and the changes in the sperm tectology in epididymis were measured on the day 8, 13, 21 and 35 of the treatment,respectively,by the method of morphological examination. The methods of immunohistochemistry and western blot were used to study the expression of heat shock protein 70(HSP70); Colorimetry was used to determine the changes in the activity and concentration of active oxygen (ROS), malondialdehyde (MDA) and antioxidized enzymes (SOD, GSH-Px, CAT); Experments of mating were used to determine the reproductive performance of female mice mated by heat stressed male mice and the fertilizing ability of the male mice. Finally, the effects of heat stress on the reproductive function of the male mice were studied systemically. And the results show as follow:1.The changes in the body weight and rectal temperature of the mice before and after the heat stress.Changes of the body weight: the body weight of mice declined before and after the heat stress, and the different value in the body weight before and after the heat stress increased with the treated temperature and decreased with the number of the heat stress.Changes in the rectal temperature: the rectal temperature of the mice rose after heat stress and the temperature increased with the treated temperature. However, there was no correlation with the number of the heat stress. The rectal temperature of the mice returned to the normal level before the next heat stress.2.The changes in the body weight, testicle and the weight exponentialThe body weight, testicle and epididymal weight of the mice after the heat stresss as compared with that of the control group and there was a negative correlation between the above declines and the treated temperature. Except the group of 37℃and 33℃treated for 8d and 13d all the rest groups of the treated mice had a significant (P<0.05) lower body weight compared with the control group;The testicle weights of the mice were significant (P<0.05) lower than the control group, except in the group of 33℃treated for 13d and 21d and 37℃treated for 13d;The epididymal weights of the mice were significant (P<0.05) lower than the control group except 33℃and 37℃更多还原