中文摘要：

目的：研究三氯生对原代大鼠卵巢颗粒细胞孕酮（P4）分泌功能的影响。方法：原代大鼠卵巢颗粒细胞培养备用。取备用的卵巢颗粒细胞采用不同浓度的三氯生（0、0．01、0．1、1μM）染毒。24h后分别采用MTT法检测颗粒细胞的相对活力、酶联免疫法（ELISA法）检测颗粒细胞P4分泌水平、实时荧光定量PCR法（qRT—PCR）及westernblot法检测类固醇激素合成急性调节蛋白（StAR）、胆固醇侧链裂解酶（P450scc）以及3β-羟基类固醇脱氢酶（3β-HSD）的基因及蛋白表达水平。结果：三氯生在本研究所采用的浓度范围内对颗粒细胞的活性并没有影响（P〉0．05）；三氯生（0．1、1μM）可抑制颗粒细胞P4的分泌，且呈现剂量依赖性下降（P〈0．05）。三氯生（0．1、1μM）可使StAR的基因表达水平显著增高、P450scc的基因表达水平下降（P〈0．05）。1μM三氯生可使StAR及P450scc的蛋白表达水平明显降低（P〈0．05）。三氯生对3B—HSD的基因及蛋白表达水平皆没有影响（P〉0．05）。结论：三氯生可抑制原代大鼠卵巢颗粒细胞的P4分泌，对类固醇激素合成关键分子的影响可能是其作用机制之一。

英文摘要：

Objective： To study the effects oftriclosan on progesterone （P4） Production in cultured rat granulosa cells. Methods： Primary cultured rat granulosa cells were prepared. Granulosa cells were treated with increasing concentrations oftriclosan （0, 0.01, 0.1 and 1 μM） for 24 h. Cellular viability was assessed by the MTT assay method. Concentrations of P4 in the medium were measured by enzyme immunoassay （ELISA）. Expression of steroidogenic acute regulatory protein （STAR）, P450 side chain cleavage enzyme （P450scc） and 3β-hydroxysteroid dehydrogenase （313-HSD） were monitored by quantative real time PCR （qRT-PCR） and western blot analysis. Results： Triclosan doses used in this study had no significant effect on granulosa cells viability （P〉0.05）. 0.1 and 1μM triclosan decreased P4 production in a dose-dependent manner （P〉0.05）. 0.1 and 1 μM triclosan significantly increased StAR mRNA expression level and reduced P450scc mRNA expression level （P〈0.05）. 1 μM triclosan reduced StAR and P450scc protein expression level （P〈0.05）. But triclosan had no significant effects on 3β-HSD both on mRNA and protein expression level （P〉0.05）. Conclusions： Triclosan can inhibit the production of P4 in primary cultured rat granulosa ceils. The disruption effects of triclosan on key steroidogenic molecules may be one of the mechanisms.