NR4A1在猪卵巢黄体中的作用探讨  

Roles of NR4A1 in porcine corpus lutea

作　　者：金鹏锦 齐丽娜[1] 魏全伟[1] 茆达干[1] 

机构地区：[1]南京农业大学动物科技学院,江苏南京210095

出　　处：《畜牧与兽医》2017年第4期1-5,共5页Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金项目(31501956);中央高校基本科研业务费专项资金;江苏省自然科学基金-青年基金项目(BK20140681)

摘　　要：通过分析NR4A1在猪卵巢黄体中的表达与定位,探讨其在猪黄体发育与退化中的潜在作用。获取猪发情周期不同阶段卵巢组织,分离黄体(早期、中期和晚期),应用放射免疫法检测孕酮和雌二醇浓度,应用蛋白免疫印迹法检测NR4A1、CYP17A1和cleaved-Caspase-3的表达,应用免疫组织化学方法检测NR4A1和cleaved-Caspase-3的定位。结果:晚期黄体(白体)的孕酮水平显著低于早期和中期,雌二醇水平显著高于早期和中期(P<0.05);早期黄体(红体)与中期黄体的NR4A1表达量无显著差异,而晚期黄体(白体)中NR4A1显著低于早期和中期(P<0.05);CYP17A1在早期黄体与中期黄体无显著差异,晚期黄体显著上升;cleaved-Caspase-3在中期黄体中表达最低,而在晚期黄体中最高(P<0.05)。此外,整个黄体发育与退化过程中,NR4A1主要在黄体类固醇生成细胞的细胞质中表达,而cleaved-Caspase-3在包括黄体类固醇生成细胞的所有细胞中均有表达。结果提示:猪卵巢黄体发育与退化过程中,NR4A1的蛋白表达趋势与孕酮水平一致,NR4A1可能参与调控猪黄体组织中孕酮的合成代谢,为深入研究NR4A1参与猪卵巢机能调控提供理论依据。Based on the natural porcine ovarian corpus luteum,the expression pattern and localization of luteal NR4A1 were analyzed to investigate the potential role during the development and regression of the corpus luteum.Porcine ovaries at different estrous cycle were collected to isolate the luteal tissues（early phase,middle phase and late phase）.Radioimmunoassay（RIA） was used to determine the progesterone and estrogen concentrations,western blot（WB） used to analyze the expression pattern of NR4A1,CYP17A1 and cleaved-Caspase-3and immunohistochemistry（IHC） applied to localize the NR4A1 and cleaved-Caspase-3.Results showed that the progesterone level of early and middle luteal tissue were lower than that in the late corpus luteum,while estrodiol were higher than that（P0.05） in late corpus luteum（corpus albicans）.In addition,the NR4A1 expression had no significant differences between early（corpus haemorrhagicum） and middle corpus luteum,which decreased significantly（P0.05） in the late corpus luteum.However,CYP17A1 expression increased significantly in the late corpus luteum,and no changes were found between the early and middle corpus luteum.Cleaved-Caspase-3 expression level was the lowest in the middle corpus luteum,while it was the highest in the late corpus luteum（P0.05）.Furthermore,the NR4A1 mainly localized in the cytoplasm of luteal steroidogenic cells,while cleaved-Caspase-3 expressed in the cytoplasm of nearly all kinds of luteal cells including steroidogenic cells.Our finding suggested that NR4A1 might be involved in the regulation of progesterone metabolism in porcine corpus lutea,providing a theoretical basis for the study of NR4A1 in the regulation of porcine ovarian functions.

关 键 词：黄体 孕酮 雌激素 NR4A1 cleaved-Caspase-3 

分 类 号：S814.1[农业科学—畜牧学]