M- CSF对人卵巢颗粒细胞表达FSH与其受体的影响及相互作用研究

摘要

目的：通过研究巨噬细胞集落刺激因子（M- CSF）对卵巢颗粒细胞表达卵泡刺激素（FSH）与其受体（FSHR）的影响及其与FSH之间的相互作用，探讨M- CSF在卵泡发育中的作用。方法体外培养人卵巢肿瘤颗粒细胞系COV434，分别给予不同浓度的M- CSF（0、5、10、25、50、100ng/ml）进行干预，24h后收取细胞和培养上清液，用荧光定量PCR法检测颗粒细胞内FSH受体表达量的变化，并用同法检测FSH对M- CSF及其受体M- CSFR的作用。用ELISA方法对上清液中细胞分泌的雌二醇（E2）进行检测，观察M- CSF对颗粒细胞分泌E2的影响。结果 M- CSF可使颗粒细胞FSHR的表达上升，呈剂量依赖性，当M- CSF浓度＞10ng/ml时，上升更为明显（P＜0.01）；FSH可以提升M- CSF及其受体在颗粒细胞内的表达，但当FSH浓度过高时（＞50ng/ml），此作用消失；M- CSF和FSH在体外条件下达到一定浓度后可以促进E2的分泌。结论 M- CSF能够促进颗粒细胞表达FSH受体和分泌E2，协助FSH发挥促卵泡发育的作用。%Objective To examine the effect of macrophage colony- stimulating factor (M- CSF) on the expression of fol i-cle stimulating hormone (FSH) and its receptor (FSHR) in human non- luteinizing granulosa cells. Methods The cultured human ovarian granulosa celltumor COV434 cells were treated with different concentrations of M- CSF (0, 5, 10, 25, 50, 100ng/ml). After 24 h, granulosa cells were col ected and the mRNA expression of FSHR, M- CSF and M- CSF receptor was detected by quantita-tive PCR. Estradiol (E2) contents in culture supernatant were measured by ELISA. Results M- CSF significantly increased the mRNA expression of FSHR in a dose dependent manner (P<0.01) , it was more markedly when the concentration of M- CSF was >10ng/ml. FSH enhanced the production of M- CSF and its receptor when the concentration<50ng/ml. Both M- CSF and FSH increased the secretion of E2 in granulosa cells. Conclusion M- CSF can increase the expression of FSHR and E2 secretion in human granulosa cell, which may promote fol icular development.