摘要
为了探讨脱氢表雄酮(DHEA)对大鼠原代卵巢颗粒细胞激素生成及相关甾体生成酶的影响,采用机械法分离培养大鼠原代卵巢颗粒细胞,放射免疫分析法检测培养液中雌二醇、孕酮和睾酮的含量;Real-time PCR法检测类固醇代谢途径中3β-羟基固醇脱氢酶(3β-HSD)、17β-羟基固醇脱氢酶(17β-HSD)、CYP450 mRNA的表达变化。结果显示:10μmol/L DHEA处理48 h后可显著促进大鼠原代卵巢颗粒细胞雌二醇(P<0.05)、孕酮(P<0.01)和睾酮的分泌量(P<0.01);显著促进3β-HSD(P<0.01)和17β-HSD(P<0.05)mRNA的表达,CYP450 mRNA表达略有降低,但无统计学意义。说明外源性DHEA处理可能通过调节3β-HSD、17β-HSD和CYP450等相关酶的基因表达,进而影响大鼠原代卵巢颗粒细胞雌激素和雄激素的分泌。
The aim of this study is to investigate the effect of dehydroepiandrosterone （DHEA） on biosynthesis of steroid hormones and cor- relative steroidogenesis enzyme mRNA expression in rat primary granulose ceils. Granulosa cells were released by mechanical method. The culture media at 48h after treatment were collected for the measurements of progesterone, estradiol and testosterone levels by radioimmunoas- say （RIA）. The mRNA expression levels of three key enzymes, such as 3β-HSD, 17β-HSD and CYP450, in the cultured granulose cells were detected by RT-PCR. Results showed that the progesterone, testosterone and estradiol levels in medium were significantly increased after treatment with 10μ mol/L of DHEA （P〈0. 01, P〈0. 05）. The treatment could enhance mRNA expression of 3β-HSD （P〈0. 01） and 17β-HSD （P〈0. 05 ）, but not affect CYP450 mRNA expression. It suggested that exogenous DHEA treatment could regulate the gene ex-pressions of 3 β-HSD, 17β-HSD and CYP450 enzymes, and then affect the secretion of estrogen and androg from cultured rat ovarian granu-losa cells.
出处
《畜牧与兽医》
北大核心
2014年第3期41-47,共7页
Animal Husbandry & Veterinary Medicine
基金
江苏省自然科学基金(BK2011647)
中央高校基本业务费(KYZ201151)
