摘要
实验一Attractin基因功能丧失的雄性小鼠细胞因子水平的变化
目的探讨敲除Attractin(Atrn)基因对雄性小鼠细胞因子水平的影响。
方法成年雄性Atrn基因敲除纯合子及正常小鼠(均为C3H/Hej种系)各12只,设为Atrn基因敲除组和对照组。内眦静脉取血法取外周血,用ELISA检测血清白细胞介素(IL) -2、白细胞介素(IL) -6及肿瘤坏死因子(TNF) -α浓度,并将结果进行组间对比。
结果Atrn基因敲除组血清IL-2浓度低于对照组雄性小鼠,血清IL-6和TNF-α浓度高于对照组雄性小鼠,差异均有统计学意义(P<0.05)。
结论Atrn基因敲除对雄性小鼠的血清IL-2浓度有下调作用,对雄性小鼠的血清IL-6和TNF-α浓度有升高作用。其具体机制有待进一步研究。
实验二Attractin及Mahognoid基因功能丧失的雄鼠下丘脑垂体睾丸等的组织学改变
目的探讨Attractin及Mahognoid基因功能丧失对雄鼠下丘脑垂体睾丸等的组织学改变。
方法取选Atrn基因功能丧失的雄鼠、Md基因功能丧失的雄鼠和正常雄鼠各6只分成3组(2个实验组,1个对照组),颈椎脱臼处死后迅速取其下丘脑、垂体、睾丸和附睾组织固定,石蜡包埋,并制做HE切片。
结果1. Atrn基因功能丧失的雄鼠和Md基因功能丧失的雄鼠下丘脑和垂体HE切片发现明显的海绵状变性。2. Atrn基因功能丧失的雄鼠和Md基因功能丧失的雄鼠睾丸和附睾HE切片未发现明显改变。
结论Atrn基因功能丧失的雄鼠和Md基因功能的丧失能引起雄鼠下丘脑和垂体的海绵状变性,但未见引起睾丸和附睾明显的组织学改变。
实验三Attractin基因功能丧失的雄性小鼠精液参数的观察
目的探讨Attractin基因功能丧失对雄鼠精液参数的改变。
方法取选正常雄鼠和Atrn基因功能丧失的雄鼠分成2组(1个对照组,1个实验组),取其中正常雄鼠和Atrn基因功能丧失的雄鼠各6只,颈椎脱臼处死小鼠后迅速取双侧睾丸、附睾和精囊腺组织,分离脂肪筋膜,滤纸吸干称重。取一侧附睾尾和部分输精管至于预先温育的0.3mlF-10培养液中,剪碎,孵育5～10min后取尽可能多的精子悬液,弃去组织,记录体积,混匀。精液常规分析,并将所得数据进行统计学分析。
结果Atrn基因功能丧失雄鼠的精子爬高速度明显下降(P<0.05),精子密度(×106个/ml)、精子活率(%)、低渗肿胀实验尾部卷曲率(%)、精子活动率(%)、睾丸、附睾和精囊腺的脏器系数(%)等其他指标未见明显改变。
结论Atrn基因功能丧失与雄鼠精子爬高速度下降具有一定相关性,但其具体机制尚有待进一步探讨。
Experiment I The change of the cytokines level of male Attractin gene knockout mice
Objective To examine the influence to the Cytokines level of Attractin gene knockout male mice.
Methods Male mature Attractin gene knockout homozygote mice and wild type mice(C3H/hej)were divided into 2 groups: the Atrn knockout group and the control group. Each group included 12 mice. Blood samples were collected from angular vein of mice. The level of IL-2,IL-6 and TNF-αwas determined with ELISA. The results would be compared between groups.
Results For the Atrn knockout group, the level of IL-2 was lower than that of the control group (P<0.05), while the level of IL-6 and TNF-αwas higher (P<0.05).
Conclusion Knocking out Atrn gene not only can down-regulate the level of IL-2 of male mice, but also can up-regulate the level of IL-6 and TNF-αof male mice. The mechanism of this phenomenon needs further study.
ExperimentⅡThe histological change of the hypothalamus-pituitary-testicle axle of Attractin gene or Mahoganoid (Md) gene knockout male mice
Objective To examine the influence to the histological HE slices of the hypothalamus-pituitary-testicle axle of Attractin gene or Mahoganoid (Md) gene knockout male mice.
Methods Male mature Attractin gene knockout homozygote mice, Male mature Md gene gene knockout homozygote mice and Male mature wild type mice(C3H/hej)were divided into 3 groups: the Atrn knockout group, the Md knockout group and the control group. Each group included 6 mice. The hypothalamus, the pituitary gland, the testicle and the epididymis of each group mice were taken out, fixed, embeded with wax and maken into HE slices.
Results 1. Obvious spongy degeneration was observed in the hypothalamus and the pituitary gland of the male mature Atrn gene gene knockout homozygote mice or male mature Md gene gene knockout homozygote mice. 2. There was no obvious change in the HE slices of the testicle and the epididymis.
Conclusion Obvious spongy degeneration was observed in the hypothalamus and the pituitary gland of the male mature Atrn gene or Md gene knockout homozygote mice. The HE slices of the testicle and the epididymis had no visible change.
ExperimentⅢThe observation of the seminal fluid parameter of Attractin (Atrn) gene knockout male mice
Objective To examine the influence to the seminal fluid parameter of Attractin (Atrn) gene knockout male mice.
Methods Male mature Atrn gene knockout homozygote mice and wild type mice(C3H/hej)were divided into 2 groups: the Atrn knockout group and the control group. Each group included 6 mice. After cervical vertebra dislocation execution, the bilateral testicle, epididymis and seminal vesicle were taken out immediately, separated from the fat and the fascia, attractted dryly by the filter paper. One side cauda epididymidis and the partial deferent duct were put into the 0.3mlF-10 nutrient fluid which was warmed in advance. Then these tissues were cut into fragments. Spermato-suspension was taken as more as possible after fostering 5~10minutes. The tissues fragments were abandoned. The volume was recorded. The seminal fluid parameter was examined, and the obtained data was eacuted a statistics analysis.
Results The sperm speed of the Atrn knockout group was lower than that of the control group (P<0.05). Compared with the control group, there was no obvious difference in the sperm density, sperm motility rate, curling tail rate of spermatozoon in the osmotic pressure swell experiment, sperm activity ratio, organ coefficient of testicle, epididymis and seminal vesicle of the Atrn knockout group.
Conclusion Knocking out Atrn gene may play an important role in the process of spermatogenisis but its specific mechanism remains to be clarified.
引文
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