THE ROLE OF PROGESTERONE IN REGULATING HUMAN GRANULOSA-CELL PROLIFERATION AND DIFFERENTIATION INVITRO
THE ROLE OF PROGESTERONE IN REGULATING HUMAN GRANULOSA-CELL PROLIFERATION AND DIFFERENTIATION INVITRO
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DOI：
10.1210/jc.76.3.696
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发表时间：
1993-03-01
影响因子：
5.8
通讯作者：
PELUSO, JJ
中科院分区：
文献类型：
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作者：
CHAFFKIN, LM;LUCIANO, AA;PELUSO, JJ
为了进一步阐明孕酮在调节颗粒细胞功能中的作用，从经历体外受精的妇女的卵泡吸出物中获得人颗粒细胞和黄体细胞并置于培养物中。以5 × 10（3）个细胞/mL接种的细胞在3天后加倍。相比之下，以50 × 10（3）个细胞/mL接种的细胞不增殖，但分化，分泌高水平的孕酮。以5 x 10（3）个细胞/mL接种的细胞并用来自以50 x 10（3）个细胞/mL接种的细胞的废培养基培养，在培养3天后数量并未增加。用RU 486（孕酮受体拮抗剂）或活性炭提取，但不能通过热灭活去除用过的培养基的生长抑制作用。在新鲜培养基中加入孕酮也能阻止细胞增殖。孕酮抑制细胞分裂的能力被RU 486或氨鲁米特减弱，这阻断了孕酮的合成。此外，表皮生长因子（EGF）刺激细胞增殖，持续暴露于孕酮阻断EGF诱导的有丝分裂。当孕酮加入2小时后，EGF，它并没有阻止EGF刺激的细胞增殖。孕酮还增加了颗粒细胞的百分比，并降低了培养3天后出现的大黄体细胞的百分比，表明孕酮抑制分化。孕酮对分化的影响是剂量依赖性的、可逆的，并且可以被hCG或8-溴-cAMP所覆盖。这些观察结果表明，孕激素直接作用于颗粒细胞通过其受体，以抑制有丝分裂和孕激素介导的抗增殖作用在2小时内的有丝分裂刺激。孕酮也阻断分化，但这种作用可以被hCG或cAMP类似物克服。这些数据表明，孕酮起着重要的作用，在控制黄体内的颗粒细胞增殖和分化，最终发展的黄体细胞的数量。
To further elucidate the role of progesterone in regulating granulosa cell function, human granulosa and luteal cells were obtained from follicular aspirates of women undergoing in vitro fertilization and placed in culture. Cells plated at 5 x 10(3) cells/mL doubled after 3 days. In contrast, cells plated at 50 x 10(3) cells/mL did not proliferate, but differentiated, secreting high levels of progesterone. Cells plated at 5 x 10(3) cells/mL and cultured with spent medium from cells plated at 50 x 10(3) cells/mL did not increase in number over 3 days of culture. The growth-inhibiting action of the spent medium was removed by either RU 486 (a progesterone receptor antagonist) or charcoal extraction, but not by heat inactivation. The addition of progesterone to fresh medium also prevented cell proliferation. Progesterone's ability to inhibit cell division was attenuated by either RU 486 or aminoglutethamide, which blocked progesterone synthesis. Further, epidermal growth factor (EGF) stimulated cell proliferation, and continuous exposure to progesterone blocked EGF-induced mitosis. When progesterone was added 2 h after EGF, it did not block EGF-stimulated cell proliferation. Progesterone also increased the percentage of granulosa cells and decreased the percentage of large luteal cells present after 3 days of culture, indicating that progesterone inhibited differentiation. Progesterone's effect on differentiation was dose dependent, reversible, and could be overridden by hCG or 8-bromo-cAMP. These observations suggest that progesterone acts directly on granulosa cells through its receptor to inhibit mitosis and that progesterone mediates its antiproliferative effects within 2 h of mitotic stimulation. Progesterone also blocks differentiation, but this effect of can be overcome by hCG or cAMP analogs. These data indicate that progesterone plays a major role in controlling the number of luteal cells that ultimately develop within a corpus luteum by regulating both granulosa cell proliferation and differentiation.