去乙酰化酶SIRT1基因在猪不同发育阶段卵泡颗粒细胞中的表达规律研究

Expression of deacetylase SIRT1 in porcine different developmental stages of follicular granulosa cells

【目的】分析去乙酰化酶SIRT1基因在猪不同发育阶段卵泡颗粒细胞中的表达规律,为进一步了解SIRT1基因在猪卵泡发育中的调控机制奠定基础。【方法】采集猪卵巢,分离卵泡颗粒细胞,根据直径将其分为≤1.5mm,〉1.5~≤3.0mm,〉3.0~≤5.0mm,〉5.0mm 4个组,利用免疫组织化学方法分析SIRT1蛋白在不同发育阶段卵泡颗粒细胞中的表达定位,利用qRT-PCR和Western blotting方法分析SIRT1基因在不同直径卵泡颗粒细胞中mRNA和蛋白的表达量。【结果】SIRT1蛋白在不同发育卵泡颗粒细胞中均有表达,随着卵泡的发育,SIRT1蛋白表达量逐渐升高,其在原始卵泡中表达量最低,在其余卵泡中表达量较高。SIRT1mRNA及其蛋白在不同直径的卵泡中表达趋势一致,在直径〉1.5~≤3.0mm卵泡中的表达量极显著高于其他直径卵泡（P〈0.01）。【结论】SIRT1基因在不同卵泡中的表达具有规律性,与猪卵巢卵泡发育具有潜在的相关性。

[Objective] The present experiment was aimed to analyze regularity of deacetylase SIRT1 expression in porcine various stages of follicles,to lay the foundation for the study of SIRT1 gene function in regulation mechanism of the porcine ovarian follicular development. [Method] Collecting porcine ovary and separating granulosa cells, granulosa cells were divided into four groups of follicular diameter ≤1.5 mm,）l. 5--43.0 mm,〉3.0--≤5.0 mm and ≤5.0 mm. Localization of SIRT1 protein in different devel- opmental follicular granulosa cells were analyzed by immunohistochemical method, expression levels of SIRT1 mRNA and protein in different size of follicular granulosa cells were analyzed by qRT-PCR and Western blotting. [Result] There had expression of SIRT1 protein in different developmental follicular granulosa cells. With follicular growth, expression of SIRT1 protein increased gradually, primordial follicles showed the lowest expression levels,other follicles were the higher expression levels. There had a same ex- pression trend of SIRT1 mRNA and protein in different diameter follicles, expression levels of SIRT1 mR-NA and protein in follicular diameter greater than 1.5 mm and less than 3.0 mm （P〈0.01）. [Conclusion] There had a some regularity of expression of deacetylase SIRT1 in different follicles,and had potential rel- evance with porcine ovarian follicular development.