丙戊酸钠对卵泡内膜细胞雄激素分泌及相关甾体合成酶改变的影响

Effect of Valprovate on Androgen Production and Correlative Steroidogenesis Enzyme mRNA Expression in Thecal Cells

目的:研究丙戊酸钠(VPA)对卵泡内膜细胞睾酮生成以及细胞色素P45017α羟化酶(CYP17)、胆固醇侧链裂解酶(P450scc)和类固醇激素急性调节蛋白(StAR)mRNA的影响。方法:采用猪的卵泡内膜细胞进行体外原代培养,并给予不同浓度的VPA(0mg/L、50mg/L、100mg/L和250mg/L)处理,48h后收集培养液,采用ELISA法检测细胞睾酮的分泌,采用荧光实时RT-PCR(TaqMan.assay)方法检测卵泡内膜细胞中CYP17、P450scc和StARmRNA表达。结果:VPA刺激了卵泡内膜细胞睾酮的分泌(P<0.05),且CYP17(P<0.01)、P450scc(P<0.01)和StAR(P<0.01)mRNA表达增加。结论:VPA可直接作用于卵泡内膜细胞,改变CYP17、P450scc和StARmRNA表达,引起雄激素的分泌变化。临床上长期服用VPA的癫痫妇女出现类似于PCOS患者的生殖内分泌功能紊乱,可能与该药物本身作用有密切联系。

Objective： To investigate the effect of valproate on testosterone and CYP 17, P450scc and StAR mRNA expression in porcine thecal cells in vitro. Methods： The thecal cells were isolated from normal porcine ovaries and cultured with DMEM-F12 （1.1） medium and treated with various concentrations of sodium valprovate （0 mg/L, 50 mg/L, 100 mg/L, 250 mg/L）. The culture media were collected after 48 h for testosterone measurements by ELISA, CYP17, P450scc and StAR mRNA in thecal cells were detected by real-time RT-PCR. Results： Valprovate caused increase of testosterone （P〈0.05）, and significantly increased of CYP17 （P〈0.01）, P450scc （P〈0.01） and StAR （P〈0.01） mRNA expression were observed in cultured cells. Conclusions： Valprovate can directly work on the thecal ceils： valprovate increased testosterone secretion by inducing CYP 17, P450scc and StAR mRNA expression in thecal cells. The occurrence of polycystic ovaries syndrome and endocrine disorders in women taking valprovate for epilepsy may be attributed to valprovate itself.