本文選題：黃體　切入點：類固醇　出處：《內(nèi)蒙古農(nóng)業(yè)大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：黃體是哺乳動物卵巢上重要的功能單位,與動物正常生殖的維持密切相關(guān),其功能性變化與動物的發(fā)情周期密切相關(guān),類固醇合成相關(guān)基因StAR、P450(CYP11A)、 3β-HSD在卵巢的周期性變化過程中具有積極作用。因此,了解綿羊卵巢周期性變化過程中黃體形成類固醇合成相關(guān)基因與潛在能量代謝通路AMPK、ERK1/2的關(guān)系對進(jìn)一步認(rèn)識卵巢功能具有價值。為此,本研究以綿羊發(fā)情周期卵巢為材料,應(yīng)用RT-qPCR、Western Blot和免疫熒光(三標(biāo))技術(shù),分析發(fā)情后第0、3、6、9、12d的綿羊卵巢中類固醇激素合成相關(guān)基因的表達(dá)變化,以期了解它們在綿羊卵巢發(fā)情周期不同時期的表達(dá)規(guī)律及能量代謝與激素合成相關(guān)基因之間潛在的聯(lián)系。結(jié)果表明,發(fā)情后第0、3、6、9、12d,綿羊卵巢黃體組織類固醇激素StAR、3β-HSD mRNA的表達(dá)量呈逐步升高然后降低的趨勢,在黃體上的表達(dá)量第9d達(dá)到最高(p0.05); 3β-HSD蛋白水平在黃體上的變化較卵巢顯著(p0.01),呈上升趨勢。而CYP11AmRNA水平隨發(fā)情周期的延續(xù)在第9到第12d仍呈逐漸升高的趨勢。同時,在已形成黃體的卵巢和未形成黃體的卵巢上GC、卵母細(xì)胞、各級卵母細(xì)胞及黃體上均有3β-HSD、StAR蛋白的表達(dá),特別指出的是(CYP17A1蛋白在卵母細(xì)胞呈陰性。且類固醇激素合成相關(guān)基因StAR、P450(CYP11A)、3p-HSD mRNA相對表達(dá)量與血液孕酮水平一致。AMPK和ERK1/2的磷酸化在發(fā)情周期的卵巢上呈逐漸升高然后下降的顯著變化。類固醇合成相關(guān)基因StAR、P450(CYP11A)、3β-HSD參與綿羊發(fā)情周期變化的進(jìn)程,且與AMPK和ERK1/2通路積極相關(guān)的實驗結(jié)果提示,類固醇激素合成相關(guān)基因與卵泡發(fā)育和黃體功能的變化密切相關(guān),該結(jié)果對進(jìn)一步認(rèn)識綿羊繁殖過程的調(diào)節(jié)機(jī)制奠定了基礎(chǔ)。
[Abstract]:Luteal body is an important functional unit in mammalian ovary, which is closely related to the maintenance of normal reproduction, and its functional changes are closely related to the oestrous cycle of animals. Steroid biosynthesis related gene Starn P450 CYP11AU, 3 尾 -HSD plays an active role in the periodic changes of ovary. It is valuable to understand the relationship between luteal steroid biosynthesis related genes and potential energy metabolism pathway AMPK ERK1 / 2 during the periodic changes of ovaries in sheep. Therefore, the ovaries of sheep estrous cycle were used as materials. The expression changes of steroid hormone synthesis-related genes in ovaries of sheep were analyzed by RT-qPCRG Western Blot and immunofluorescence (triple labeling) technique on the 12th day after oestrus. In order to understand their expression at different stages of oestrous cycle in sheep and the potential relationship between energy metabolism and hormone biosynthesis genes, the results showed that, On the 12th day after oestrus, the expression of steroid steroid hormone Starn3 尾 -HSD mRNA in ovarian luteal tissue of sheep increased gradually and then decreased. The expression level of 3 尾 -HSD on the corpus luteum reached the highest level on the 9th day, and the level of 3 尾 -HSD protein in the corpus luteum was significantly higher than that in the ovary, and the level of CYP11AmRNA increased gradually with the continuation of the estrus cycle from the 9th to the 12th day. At the same time, the expression of 尾 -HSD protein in the corpus luteum was significantly higher than that in the ovary. The expression of 3 尾 -HSD-Star protein was observed in the ovaries of luteal formation and ovaries without luteal formation, and in oocytes, oocytes and corpus luteum. It was pointed out that CYP17A1 protein was negative in oocytes, and the relative expression of steroid hormone synthesis-associated gene StARP450, CYP11Af3p-HSD mRNA was consistent with the level of blood progesterone. Phosphorylation of AMPK and ERK1/2 increased gradually in oocytes during estrous cycle and then decreased. The steroid biosynthesis related gene Starn P450, CYP11Af3 尾 -HSD, was involved in the process of estrous cycle change in sheep. The results of positive correlation with AMPK and ERK1/2 pathway suggested that steroid hormone biosynthesis related genes were closely related to follicular development and luteal function, which laid a foundation for further understanding the regulation mechanism of sheep reproductive process.
【學(xué)位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S826

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