Magoffin D A, Weitsman S R
Department of Obstetrics and Gynecology, Cedars-Sinai Research Institute Cedars-Sinai Medical Center/UCLA School of Medicine 90048.
Biol Reprod. 1993 May;48(5):1166-73. doi: 10.1095/biolreprod48.5.1166.
Evidence accumulating in the literature supports the hypothesis that insulin-like growth factor-I (IGF-I) may play a role in stimulating differentiation of the ovarian theca-interstitial cells (TIC) during early follicular development. IGF-I has been shown to synergistically enhance the stimulation of androgen biosynthesis and to increase LH binding in TIC. The purpose of the present studies was to examine the role of IGF-I in TIC differentiation by determining the effects of IGF-I on 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) mRNA expression in TIC stimulated to differentiate in vitro. TIC were isolated from the ovaries of hypophysectomized immature rats by Percoll gradient centrifugation and cultured in the presence and absence of LH and IGF-I for up to 6 days. At various times cytoplasmic RNA was extracted from the TIC, and 3 beta-HSD mRNA was measured by specific assay using reverse transcription followed by the polymerase chain reaction. Amplification of 3 beta-HSD mRNA using primers designed to distinguish between the type I and type II 3 beta-HSD gene products revealed that the TIC expressed primarily the type I gene. Increasing concentrations of LH (0-1 microgram/ml) stimulated a dose-related increase in 3 beta-HSD mRNA that was approximately 3-fold at 100 ng/ml of LH. Addition of IGF-I (30 ng/ml) increased 3 beta-HSD mRNA approximately 2-fold over TIC treated with LH alone.(ABSTRACT TRUNCATED AT 250 WORDS)
文献中积累的证据支持这样一种假说,即胰岛素样生长因子-I(IGF-I)可能在卵泡早期发育过程中刺激卵巢膜间质细胞(TIC)分化方面发挥作用。IGF-I已被证明能协同增强雄激素生物合成的刺激作用,并增加TIC中促黄体生成素(LH)的结合。本研究的目的是通过确定IGF-I对体外诱导分化的TIC中3β-羟基类固醇脱氢酶(3β-HSD)mRNA表达的影响,来研究IGF-I在TIC分化中的作用。通过Percoll梯度离心从垂体切除的未成熟大鼠卵巢中分离出TIC,并在有和没有LH及IGF-I的情况下培养长达6天。在不同时间从TIC中提取细胞质RNA,使用逆转录随后进行聚合酶链反应的特异性测定法来测量3β-HSD mRNA。使用设计用于区分I型和II型3β-HSD基因产物的引物对3β-HSD mRNA进行扩增,结果显示TIC主要表达I型基因。LH浓度增加(0 - 1微克/毫升)刺激3β-HSD mRNA呈剂量相关增加,在100纳克/毫升LH时约增加3倍。添加IGF-I(30纳克/毫升)使3β-HSD mRNA比仅用LH处理的TIC增加约2倍。(摘要截短于250字)