促黄体素(LH)对猪卵巢颗粒细胞细胞周期及类固醇激素分泌的影响  

Effects of luteinizing hormone on cell cycle and steroid secretion of porcine granulosa cells

作　　者：王伟[1] 贺彧[1] 张海燕[1] 田宽校[1] 宋晓光[1] 王晶晶[1] 徐银学[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：《江苏农业学报》2011年第1期105-109,共5页Jiangsu Journal of Agricultural Sciences

基　　金：国家高技术研究发展计划项目(2006AA10Z136)

摘　　要：为揭示促黄体素对猪卵巢颗粒细胞生长及类固醇分泌的影响,从当地屠宰场采集猪卵巢,选择表面直径大于6 mm的卵泡(优势卵泡)作为试验材料,将从其中获得的颗粒细胞进行体外培养,并在培养基中添加不同浓度(0 IU/ml、2 IU/ml和4 IU/ml)的促黄体素(LH),通过流式细胞仪、放射性免疫和定量PCR分别检测细胞周期的情况、类固醇激素的分泌和相关基因mRNA的表达。结果显示:高剂量的LH(4 IU/ml)可使G0/G1期的细胞数量由92.79%减少到86.12%;S期细胞数量由1.97%增加到8.43%,并能显著提高细胞周期依赖性蛋白激酶-4(CDK4)和细胞周期素-D2(Cyclin D2)的表达水平;高剂量的LH(4 IU/ml)能显著减少培养基中E2和P4的浓度,并能降低芳香化酶(P450arom)和胆固醇侧链裂解酶(P450scc)的表达。结果表明LH对猪卵巢颗粒细胞生长和类固醇激素分泌有调控作用。To investigate the effect of luteinizing hormone（LH） on growth and steroid secretion of porcine granulosa cells,dominant follicles from porcine ovaries collected from a local slaughterhouse were selected based on the diameter（6 mm）.Granulosa cells（GCs） obtained from porcine dominant follicles were cultured in the medium supplemented with different doses of LH（0 IU/ml,2 IU/ml and 4 IU/ml）.Cell cycle and steroid secretion were analyzed by flow cytometry,RIA and real-time fluorescence quantitative PCR,respectively.The results showed that the high dose of LH（4 IU/ml） significantly decreased the percentage of GCs from 92.79% to 86.12% at G0/G1 stage,increased the percentage of GCs from 1.97% to 8.43% at S stage,and significantly improved the expression of cyclin-dependent kinase-4（CDK4） and Cyclin D2.The high dose of LH（4 IU/ml） significantly lowered the concentrations of estradiol（E2） and progesterone（P4） in the medium,and down-regulated the expression of P450 aromatase（P450arom） as well as cholesterol side-chain cleavage enzyme P450（P450scc）.The results indicate that LH regulates the growth and steroid secretion of porcine granulosa cells.

关 键 词：猪 颗粒细胞 促黄体素 细胞周期 类固醇激素 基因表达 

分 类 号：S828[农业科学—畜牧学]