Wang Hongbo, Wen Yan, Polan Mary Lake, Boostanfar Robert, Feinman Michael, Behr Barry
Huntington Reproductive Center, Westlake Village, CA 91361, USA.
Hum Reprod. 2005 Oct;20(10):2749-56. doi: 10.1093/humrep/dei123. Epub 2005 Jun 15.
Expression and selective cellular localization of granulocyte-macrophage colony-stimulating factor (GM-CSF) and its receptor in ovarian tissue imply an autocrine/paracrine role in ovarian function. Evidence indicating a functional role for GM-CSF in ovarian follicular cell function has been provided by studies with GM-CSF knockout (GM-/-) mice, which suggest that GM-CSF influences events associated with murine follicular maturation.
Immature female rats were treated with GM-CSF, FSH or saline for 5 or 10 days. Ovaries were collected for histologic examination and immunostaining determination of CYP17, a theca cell marker. In addition, ovarian section slides were evaluated by immunofluorescence for CD45, an ovarian leukocyte marker. To investigate the possible mechanism of GM-CSF action on follicular development, theca-interstitial cells (T-I) were separated and cultured. Cells were treated with increasing concentrations of GM-CSF, then evaluated for CYP17 mRNA and protein expression assays.
After 10 days of treatment with GM-CSF, the number of small preantral and large preantral follicles was significantly increased compared with the control group (P < 0.05). Similarly, treatment with FSH increased the number of small preantral and large preantral follicles (P < 0.05). CD45 expression measured by immunofluorescence was not different in the three groups, indicating that the distribution of leukocytes was unchanged. In addition, CYP17 was increased in the T-I cells both in vivo and in vitro after GM-CSF treatment.
The present results suggest that GM-CSF may play a significant role in follicular development.
粒细胞-巨噬细胞集落刺激因子(GM-CSF)及其受体在卵巢组织中的表达和选择性细胞定位表明其在卵巢功能中具有自分泌/旁分泌作用。对GM-CSF基因敲除(GM-/-)小鼠的研究提供了证据,表明GM-CSF在卵巢卵泡细胞功能中具有功能性作用,提示GM-CSF影响与小鼠卵泡成熟相关的事件。
将未成熟雌性大鼠用GM-CSF、促卵泡激素(FSH)或生理盐水处理5天或10天。收集卵巢进行组织学检查,并通过免疫染色测定卵泡膜细胞标志物CYP17。此外,通过免疫荧光评估卵巢切片载玻片上的卵巢白细胞标志物CD45。为了研究GM-CSF对卵泡发育作用的可能机制,分离并培养卵泡膜-间质细胞(T-I)。用浓度递增的GM-CSF处理细胞,然后评估CYP17 mRNA和蛋白表达检测结果。
用GM-CSF处理10天后,与对照组相比,小窦前卵泡和大窦前卵泡的数量显著增加(P < 0.05)。同样,用FSH处理也增加了小窦前卵泡和大窦前卵泡的数量(P < 0.05)。通过免疫荧光测定的CD45表达在三组中无差异,表明白细胞分布未改变。此外,GM-CSF处理后,体内和体外T-I细胞中的CYP17均增加。
目前的结果表明GM-CSF可能在卵泡发育中起重要作用。