本文關(guān)鍵詞： 硼 大鼠 睪丸 脾臟 豬卵泡顆粒細胞 ER 增殖與凋亡　出處：《中國農(nóng)業(yè)科學院》2016年博士后論文　論文類型：學位論文

【摘要】：硼是人和動物不可缺少的微量元素之一,硼缺乏可致動物胚胎發(fā)育受阻、生殖和免疫功能出現(xiàn)異常,適量補硼對機體生殖和免疫功能有不同程度促進作用,高劑量硼則產(chǎn)生損傷甚至毒性作用。目前,硼影響人和動物生殖和免疫功能的作用機制尚不清楚。本研究通過體內(nèi)和體外實驗,研究不同劑量硼對大鼠睪丸ER表達與定位,脾臟淋巴細胞亞群與細胞因子分泌,以及睪丸、脾臟和體外培養(yǎng)豬卵泡顆粒細胞增殖與凋亡的影響,從不同層面揭示硼影響機體生殖和免疫功能的作用機制,以期為硼在動物生產(chǎn)和人類營養(yǎng)中的合理應(yīng)用提供科學依據(jù)。研究體內(nèi)試驗選用剛斷乳SPF級雄性SD大鼠為動物,體外試驗選用豬卵泡顆粒細胞為研究對象。128只大鼠分為8組,分別飲用添加0、10、20、40、80、160、320和640mg/L硼的蒸餾水,體外分離培養(yǎng)豬卵泡顆粒細胞并分別添加0.01、0.1、0.5、1.0、2.0、50和100mmM硼。結(jié)果如下：飲水添加20和40 mg/L硼可顯著增加60d和90d大鼠睪丸內(nèi)ER含量和ER a陽性細胞率,促進ER α mRNA表達；顯著升高90d大鼠血清IgG水平、脾臟INF-β和IL-4含量及CD3和CD4陽性細胞數(shù)；極顯著促進90d大鼠睪丸和脾臟PCNA表達,抑制Caspase-3表達,降低CD3+T細胞中Caspase-3陽性細胞數(shù)。添加320和640m∥L硼則不同程度降低60d和90d大鼠睪丸內(nèi)ER含量和ERa陽性細胞率,抑制ER α mRNA表達,明顯降低90d大鼠血清IgG含量、脾臟INF-β和IL-4含量及CD3和CD4陽性細胞數(shù)；降低90d大鼠睪丸和脾臟PCNA陽性細胞數(shù),增加Caspase-3陽性細胞數(shù)。體外細胞培養(yǎng)添加0.01、0.1、0.5和1.0mM硼可促進豬卵泡顆粒細胞增殖,降低其晚期凋亡率和死亡率；硼添加量高于2.0mM時,則可致豬卵泡顆粒細胞皺縮,胞核分解,早、晚期細胞凋亡率及死亡率顯著增加。研究證明ER-α在硼影響睪丸發(fā)育和精子發(fā)生中起著重要作用,初步揭示細胞增殖和凋亡可能是硼影響機體生殖和免疫功能的重要機制。研究首次發(fā)現(xiàn),適量添加硼(硼添加量0.01-1.0 mM)對體外培養(yǎng)豬卵巢顆粒細胞的生長有明顯促進作用,對卵泡顆粒細胞的凋亡有明顯抑制作用；高硼(添加量大于2.0mM)則對豬卵泡顆粒細胞生長有明顯不良影響甚至毒性作用,加速卵泡顆粒細胞凋亡的發(fā)生,細胞死亡數(shù)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 偽 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 偽 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- 偽 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.
【學位授予單位】：中國農(nóng)業(yè)科學院
【學位級別】：博士后
【學位授予年份】：2016
【分類號】：S816

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本文編號：1445657