Effects Of Progesterone On Progesterone Receptor Expression In Cultured Endometrium Stromal Cells From Bovine

Progesterone receptor (PR) is a member of nucleolus-receptor superfamily. PR exert important roles for ovulation,embryo implantation, mammary gland development: and other reproductive process.In female animal physiological level,progesterone inhibit PR expression in endometrium. In vitro endometrium,is it possible to inhibit PR expression? The aim of this study is to discuss the effects of progesterone on progesterone receptor expression in cultured endometrium stromal cells from bovine.In the experiment,firstly,endometrium cells were cultured in FBS-free medium and then endometrium cells were treated with different concentration of progesterone. Secondly,after being treated different time,endometrium cells were collected and extracted total RNA. Finally,PR expression level were assessed by semi-quantitative PCR under β-actin reference. The mothods and results were as follow:1.Primary culture of bovine endometrium cells was carried out by tissue-culture method,the culture system was:DMEM/F₁₂+FBS(20% for primary culture, 10% for subculture)+ 100 IU/ml penisillin and streptomycin,and incubated at 37℃ in a 5% CO₂ humidified atmosphere CO₂ Water jacketed incubator.2.Mix the cells during3 8generations from 8 nonepregnant luteal-term scalper.Cultured the cells till covered 60% of the culture bottle bottom, changed the culture medium to FBS-free DMEM/F₁₂,After 24h,treated with different concentration of progesterone(50, 100> 200, 400ng/L) for different time.Then collected the cells for RNA extracting.3.Semi-quantitative RT-PCR was used in this study.Firstly,adjust the template...