巨噬细胞集落刺激因子在卵泡黄素化颗粒细胞中的表达

Expression of macrophage colony-stimulating factor in human luteinized granulosa cells

目的 了解巨噬细胞集落刺激因子 (M CSF)在人卵泡黄素化颗粒细胞中的表达 ,探讨促卵泡激素 (FSH)在体外对人卵泡黄素化颗粒细胞合成分泌M CSF的调节作用。方法 取接受卵母细胞单精子显微注射治疗的 2 0例妇女的卵泡黄素化颗粒细胞 ,采用免疫细胞化学染色法测定黄素化颗粒细胞中M CSF的表达 ,并将黄素化颗粒细胞单独培养及在不同浓度的FSH(2、5、15、2 5IU/ml)作用下培养 72h ,采用酶联免疫吸附法测定黄素化颗粒细胞培养液中的M CSF含量。结果 约 80 %的黄素化颗粒细胞胞浆有M CSF表达 ;黄素化颗粒细胞在培养 72h后 ,基础状态下M CSF分泌量为(5 9± 13)ng/L ,浓度为 2、5、15、2 5IU/ml的FSH作用下M CSF分泌量分别为 (16 5± 32 )、(2 10± 5 8)、(2 99± 6 6 )、(390± 78)ng/L ,在不同浓度的FSH作用下黄素化颗粒细胞培养液中M CSF的含量与基础状态下相比 ,差异均有极显著性 (P <0 0 1)。结论 卵泡黄素化颗粒细胞能合成分泌M CSF ;FSH能促进黄素化颗粒细胞M CSF的分泌 ,FSH对卵泡发育和成熟的调节 ,可能部分通过M

Objective To detect and localize the expression of macrophage colony stimulating factor (M CSF) in human luteinized granulosa cells and investigate the effects of follicle stimulating hormone (FSH) on M CSF production by human luteinized granulosa cells in vitro Methods Human luteinized granulosa cells were isolated from follicular fluid of superovulated infertile patients undergoing intracytoplasmatic sperm injection Some of the luteinized granulosa cells were used for detecting M CSF by immunocytochemical staining (ABC method) Most of them were cultured with HAM′s F 10 medium alone or plus various concentrations of FSH (2, 5, 15, 25 IU/ml) The media were collected after 72 hours and their M CSF contents were measuered by a solid phase enzyme linked immunosorbant assay Results About 80% of the luteinized granulosa cells were positively stained for M CSF antibody The immunoreactive signals of M CSF were specially located in the cytoplasma of the luteinized granulosa cells No immunoreactivity of M CSF was observed in vaginal squamous epithelial cells through contaminations during transvaginal oocyte retrieval The concentration of M CSF in cultured luteinized granulosa cells medium without FSH stimulation was low However, addition of 2, 5, 15, 25 IU/ml concentrations of FSH caused significant dose dependent increases of M CSF production after 72 hours culture by luteinized granulosa cells ( P <0 01) Conclusions Human luteinized granulosa cells can synthesize and secrete M CSF FSH can directly stimulate M CSF secretion by human luteinized granulosa cells The mechanism of FSH regulating follicular development may partially via M CSF/receptor pathway