丙戊酸钠对人卵巢黄素化颗粒细胞激素分泌及相关甾体合成酶mRNA影响  

Effects of valproate on steroid production and correlative steroidogenesis enzyme mRNA in human luteinized granulose cells

作　　者：朱珠[1] 孙国栋[1] 刘嘉茵[1] 

机构地区：[1]南京医科大学第一附属医院妇产科,南京210009

出　　处：《中国药理学通报》2004年第6期628-632,共5页Chinese Pharmacological Bulletin

基　　金：国家科技部重点"973"项目资助课题;No 2 0 0 2CB5 12 90 8

摘　　要：目的　丙戊酸钠 (VPA)为抗癫痫的有效药物 ,长期应用可引起类似于多囊卵巢综合征 (PCOS)生殖内分泌功能紊乱 ,本研究通过人黄素化卵巢颗粒细胞原代培养体系 ,研究VPA对黄素化颗粒细胞的激素生成及类固醇激素生成急性调节蛋白 (StAR)、胆固醇侧链裂解酶 (P4 5 0scc)和芳香化酶(P4 5 0arom)的mRNA影响。方法　收集体外受精 -胚胎移植 (IVF ET)时的人黄素化颗粒细胞进行体外培养 ,适时给于不同浓度的VPA(0、10 0、2 5 0mg·L-1)处理 ,2d后收集培养液、细胞 ,采用放射免疫方法测定颗粒细胞分泌孕酮及雌二醇的量 ,并以细胞蛋白浓度校正激素含量。采用荧光实时RT PCR (TaqMan assay)方法检测颗粒细胞中StAR、P4 5 0scc和P4 5 0arommRNA表达。结果　VPA刺激人黄素化颗粒细胞的孕酮 (P <0 0 5 )和雌二醇 (P <0 0 1)的分泌 ,且StARmRNA (P <0 0 1)、P4 5 0scc(P <0 0 5 )和P4 5 0arom(P <0 0 1)表达增加。结论　VPA可直接作用于黄素化颗粒细胞 ,通过改变StAR、P4 5 0scc和P4 5 0arommRNA的表达 ,而引起孕酮和雌二醇的分泌变化 ,而临床上长期服用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 ,同时提示了非遗传因素即环境因素在PCOS发病中也发挥一定作用。AIM Valproate (VPA) is widely used to treat epilepsy. Long-term treatment of women with VPA has been reported to be associated with a PCOS-like syndrome. The aim of the present study was to investigate the effects of clinically relevant concentrations of valproate on steroidogenesis and steroidogenesis acute regulatory protein (StAR),cholesterol side-chain cleavage enzyme (P450scc)and cytochrome P450 aromatase (P450arom) mRNA expression in human luteinized granulosa cells in vitro. METHODS Human luteinized granulosa cells were isolated from oocyte retrieval of in vitro fertilization procedure and were cultured with DMEM medium and treated with various concentrations of valproate (0, 100, 250 mg·L -1), the culture media was collected after 2 days for progesterone and estradiol measurements by standard radiommunoassay, StAR, P450scc and P450arom mRNA in granulose cells were detected by fluorescent real-time RT-PCR (TaqMan assay). RESULTS Valproate caused significant increase of progesterone (P<0.05) and estradiol(P<0.01) production, and significantly increasein StAR (P<0.01), P450scc(P<0.05) and P450arom (P<0.01) mRNA expression were observed in cultured cells. CONCLUSION Valproate directly altered steroidognesis of steroidogenesis cells: valproate increases progesterone and estradiol secretion by inducing StAR, P450scc and P450arom mRNA expression in human luteinized granulosa cell. And the occurrence of polycystic ovaries and endocrine disorders in women taking valproate for epilepsy may be attributed to valproate itself. It also implies that environmental chemicals may be related to the pathogenesis of PCOS.

关 键 词：丙戊酸钠 卵巢黄素化颗粒细胞 甾体激素生成 甾体生成酶 MRNA表达 

分 类 号：R322.65[医药卫生—人体解剖和组织胚胎学] R342.22[医药卫生—基础医学]