微量元素硼影响动物生殖和免疫功能的作用机理研究

发布时间：2018-01-19 21:51

本文关键词： 硼大鼠睾丸脾脏猪卵泡颗粒细胞 ER 增殖与凋亡　出处：《中国农业科学院》2016年博士后论文　论文类型：学位论文

【摘要】：硼是人和动物不可缺少的微量元素之一,硼缺乏可致动物胚胎发育受阻、生殖和免疫功能出现异常,适量补硼对机体生殖和免疫功能有不同程度促进作用,高剂量硼则产生损伤甚至毒性作用。目前,硼影响人和动物生殖和免疫功能的作用机制尚不清楚。本研究通过体内和体外实验,研究不同剂量硼对大鼠睾丸ER表达与定位,脾脏淋巴细胞亚群与细胞因子分泌,以及睾丸、脾脏和体外培养猪卵泡颗粒细胞增殖与凋亡的影响,从不同层面揭示硼影响机体生殖和免疫功能的作用机制,以期为硼在动物生产和人类营养中的合理应用提供科学依据。研究体内试验选用刚断乳SPF级雄性SD大鼠为动物,体外试验选用猪卵泡颗粒细胞为研究对象。128只大鼠分为8组,分别饮用添加0、10、20、40、80、160、320和640mg/L硼的蒸馏水,体外分离培养猪卵泡颗粒细胞并分别添加0.01、0.1、0.5、1.0、2.0、50和100mmM硼。结果如下：饮水添加20和40 mg/L硼可显著增加60d和90d大鼠睾丸内ER含量和ER a阳性细胞率,促进ER α mRNA表达；显著升高90d大鼠血清IgG水平、脾脏INF-β和IL-4含量及CD3和CD4阳性细胞数；极显著促进90d大鼠睾丸和脾脏PCNA表达,抑制Caspase-3表达,降低CD3+T细胞中Caspase-3阳性细胞数。添加320和640m∥L硼则不同程度降低60d和90d大鼠睾丸内ER含量和ERa阳性细胞率,抑制ER α mRNA表达,明显降低90d大鼠血清IgG含量、脾脏INF-β和IL-4含量及CD3和CD4阳性细胞数；降低90d大鼠睾丸和脾脏PCNA阳性细胞数,增加Caspase-3阳性细胞数。体外细胞培养添加0.01、0.1、0.5和1.0mM硼可促进猪卵泡颗粒细胞增殖,降低其晚期凋亡率和死亡率；硼添加量高于2.0mM时,则可致猪卵泡颗粒细胞皱缩,胞核分解,早、晚期细胞凋亡率及死亡率显著增加。研究证明ER-α在硼影响睾丸发育和精子发生中起着重要作用,初步揭示细胞增殖和凋亡可能是硼影响机体生殖和免疫功能的重要机制。研究首次发现,适量添加硼(硼添加量0.01-1.0 mM)对体外培养猪卵巢颗粒细胞的生长有明显促进作用,对卵泡颗粒细胞的凋亡有明显抑制作用；高硼(添加量大于2.0mM)则对猪卵泡颗粒细胞生长有明显不良影响甚至毒性作用,加速卵泡颗粒细胞凋亡的发生,细胞死亡数目增多。
[Abstract]:Boron is one of the indispensable trace elements in human and animal. Boron deficiency can hinder the development of animal embryos and lead to abnormal reproductive and immune function. Proper amount of boron supplementation can promote the body's reproductive and immune function to varying degrees. At present, the mechanism of the effect of boron on reproductive and immune function of human and animal is not clear. This study is carried out in vivo and in vitro. To study the effects of different doses of boron on ER expression and localization in rat testis, lymphocyte subsets and cytokines secretion in spleen, proliferation and apoptosis of testis, spleen and porcine follicular granulosa cells in vitro. The mechanism of the effect of boron on reproductive and immune function was revealed from different aspects. In order to provide a scientific basis for the rational application of boron in animal production and human nutrition, the male SD rats of newly weaned SPF grade were selected as animals in vivo. Porcine follicular granulosa cells were selected as the object of study in vitro. 128 rats were divided into 8 groups. Porcine follicular granulosa cells were isolated from distilled water of 320 mg / L and 640 mg / L boron in vitro and supplemented with 0.01mg / L 0.51% 0.51.0C 2.0, respectively. The results were as follows: adding boron for 20 and 40 mg/L in drinking water significantly increased ER content and ER a positive cell rate in testis of 60 d and 90 d rats. Promote the expression of ER 伪 mRNA; The levels of serum IgG, the contents of INF- 尾 and IL-4 in spleen and the number of CD3 and CD4 positive cells were significantly increased in 90 d rats. The expression of PCNA in testis and spleen of 90 day rats was significantly increased, and the expression of Caspase-3 was inhibited. Reduce CD3. The number of Caspase-3 positive cells in T cells: the ER content and ERa positive cell rate in testis of 60 d and 90 d rats were decreased by adding 320 and 640 m / L boron. The expression of ER 伪 mRNA was inhibited, the content of serum IgG, the contents of INF- 尾 and IL-4 in spleen and the number of CD3 and CD4 positive cells in spleen of 90 d rats were significantly decreased. The number of PCNA positive cells and the number of Caspase-3 positive cells in testis and spleen of 90 d rats were decreased and the number of Caspase-3 positive cells were increased. 0.5 and 1.0 mm boron could promote the proliferation of porcine follicular granulosa cells and reduce the late apoptosis rate and mortality. When the addition of boron was more than 2.0 mm, the granulosa cells of pig follicles shrank and the nucleus decomposed early. The rate of late apoptosis and death rate were significantly increased. Studies have shown that ER- 伪 plays an important role in the effects of boron on testicular development and spermatogenesis. It is revealed that cell proliferation and apoptosis may be the important mechanism of boron affecting reproductive and immune function. The addition of boron (0.01-1.0 mm) significantly promoted the growth of porcine ovarian granulosa cells in vitro and inhibited the apoptosis of follicular granulosa cells. High boron (more than 2.0 mm) had obvious adverse effects on the growth of porcine follicular granulosa cells, and accelerated the occurrence of follicular granulosa cell apoptosis and increased the number of cell death.
【学位授予单位】：中国农业科学院
【学位级别】：博士后
【学位授予年份】：2016
【分类号】：S816

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