4-氨基比啶对人黄素化颗粒细胞孕酮分泌的抑制作用被引量：1

Inhibiting effect of 4-aminopyridine on progesterone production in human ovarian luteinized granulosa cells

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摘要目的探讨钾通道阻滞剂4-氨基比啶(4-AP)对人黄素化颗粒细胞孕酮分泌的影响。方法收集行体外受精-胚胎移植患者的卵巢黄素化颗粒细胞,分别加入不同浓度的4-AP(1、5、10mmol/L),检测其对于基础状态(HCG=0IU/L)及HCG诱导状态下(HCG=1250IU/L)孕酮分泌的影响;以不加4-AP的细胞作为对照。采用放射免疫测定法(RIA)检测细胞内cAMP浓度;应用RT-PCR及Western blotting分析颗粒细胞类固醇激素合成急性调控蛋白(StAR)mRNA及蛋白表达的变化;采用MTT比色法检测不同浓度4-AP对颗粒细胞增殖的抑制作用。结果基础状态和HCG诱导状态下,4-AP各浓度组人黄素化颗粒细胞孕酮及cAMP水平均较对照组降低(P<0.01)。随着4-AP浓度的增加,StAR mRNA及蛋白表达逐渐降低(P<0.01),对颗粒细胞增殖的抑制作用逐步增强(P<0.01),呈浓度依赖性。结论4-AP能够抑制人黄素化颗粒细胞基础状态和HCG诱导状态下孕酮的分泌;4-AP可能通过降低颗粒细胞cAMP的生成和StAR的表达并抑制其增殖发挥作用。 Objective To explore the effects of K^＋ channel blocker 4-aminopyridine（4-AP）on the progesterone production in human ovarian luteinized granulosa cells. Methods Ovarian luteinized granulosa cells were recovered from women with regular menses who underwent in vitro fertilization programme. Luteinized granulosa cells were co-incubated with various concentrations of 4-AP （1,5, 10 mmol/L） to examine the effects on progesterone production under basal condition（ HCG = 0 IU/L） and HCG-induced condition（HCG = 1 250 IU/L）. Those without co-incubation with 4-AP were served as controls. Cyclic adenosine monophosphate （cAMP） levels were evaluated by radioimmunoassay （RIA）, RT-PCR and Western Blotting were employed to detect the expression of steroidogenic aetue regulatory protein （STAR） mRNA and protein, and the inhibition of cell proliferation was assessed using methyl thiazolyl tetrazolium （ MTT ） assay. Results Under basal condition and HCG-induced condition, the levels of progesterone and cAMP in luteinized granulosa cells in groups of various concentrations of 4-AP were significantly lower than those in control group （ P 〈 0.01 ）. 4-AP decreased the expression of StAR mRNA and protein, and inhibited the proliferation of luteinized granulosa cells in a dose-dependent manner （P 〈 0. 01）. Conclusion 4-AP can inhibit the progesterone production in luteinized granulosa cells under basal and HCG- induced conditions, which may be realized by decreasing cAMP generation, reducing StAR mRNA and protein expression and inhibiting cell proliferation.

作者崔娜赵志明吴小华徐素欣郝桂敏曹金凤

机构地区河北医科大学第二医院生殖医学科河北医科大学外科学教研室

出处《上海交通大学学报（医学版）》 CAS CSCD 北大核心 2009年第1期42-46,共5页 Journal of Shanghai Jiao tong University:Medical Science

关键词 4-氨基吡啶颗粒细胞类固醇激素合成急性调控蛋白孕酮 4-aminopyridine granulosa cells steroidogenic acute regulatory protein progesterone

分类号 R96 [医药卫生—药理学] R711.6 [医药卫生—妇产科学]

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