氯丙嗪对大鼠卵巢颗粒细胞分泌功能的影响

Effect of chlorpromazine on the steroidogenesis of rat ovarian grandlose cells in vitro

为研究大鼠卵巢颗粒细胞(GC)的分泌功能,用氯丙嗪染毒体外培养的大鼠GC,采用MTT法检测细胞相对活力,ELISA法检测收集培养液中孕酮(P)和雌二醇(E2)的含量,半定量RT-PCR检测激素分泌相关调控基因FSHR、StAR、P450scc和P450arom mRNA的表达,免疫细胞化学染色法检测细胞中特异卵泡刺激素受体的表达。结果表明:经0.1、1.0和10.0μmol/L氯丙嗪染毒24 h后,与对照组相比,细胞相对活力分别为87.95%、83.96%和74.48%,E2的分泌量和StAR mRNA相对表达水平分别从对照组的6.16 pg/μg、2.014显著下降到3.70 pg/μg、0.311,呈现明显的剂量?效应关系,表明转运蛋白StAR可能是氯丙嗪影响颗粒细胞激素分泌的关键位点之一。

To study the toxic effects and explore the possible mechanism of chlorpromazine exposure to rat ovarian granulosa cell steroidogenesis in vitro, immature ovarian granulosa cells of wistar rat was primary cultured. After exposure to chlorpromazine for 24 h, cell viability was detected by MTT assay, content of progesterone （P） and estradiol （E2） collected from medium were detected by ELISA assay; FSHR, STAR, P450scc and P450arom mRNA expression were detected by semi-quantitative RT-PCR, and expression of follicle stimulating hormone receptor-specific receptor（FSHR） in granulosa cells was detected by immunocytochemical staining assay. The results showed that chlorpromazine could significantly decrease the viability of rat ovarian granulosa cells, thus affect steroidogenesis and StAR maybe one of the key factors to affect the steroidogenesis of granulosa cells.