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Involvement of Macrophage Colony- stimulating Factor( M- CSF) in the Function of Follicular Granulosa Cells and Its Molecular Mechanism
Author(s): Zhang Wei, Xu Song, Zhang Zhifen, Department of Gynecology, Hangzhou First People’s Hospital, Nanjing Medical University
Pages: 100-
105
Year: 2016
Issue:
1
Journal: Journal of Medical Research
Keyword: M-CSF; FSH; Granulosa Cells(GCs); Estradiol;
Abstract: Objective To investigate the effect of macrophage colony- stimulating factor( M- CSF) in follicular granulosa and its molecular mechanism. Mehtods Human luteinized granulosa cells( LCC) were isolated from follicular fluid of superovulated infertile patients( average age 30. 8 ± 2. 1years old) undergoing in vitro fertilization and embryo transfer. LCC were cultured with DMEM / F12 medium plus various concentration of M- CSF( 0,10,25,50,100 ng / ml),M- CSF + Letrozole( 10- 6mol / L),FSH( 0,10,25,50,100 IU / m L),FSH + Letrozole( 10- 6mol / L). Media were collected at 24 hours after culture and estradiol( E2) in media were measured by ELISA. Expression levels of FSH receptor and M- CSF receptor were deteced by quantiative RT- PCR. After cultured COV434 were treated with M- CSF,cell proliferation was quantified by MTs assay and protein expression was detected by Western blot. Results Either M- CSF or FSH stimulated the production of E2. The production of FSH receptors was enhanced by M- CSF or M- CSF + Letrozole in vitro in a dose- dependent manner. Conversely,FSH was able to promote the expression of the receptor of M- CSF when present in an appropriate concentration. With the increasing of M- CSF concentration,the cell proliferation of COV343 showed an upward trend. M- CSF induced phosphorylation of JNK and P38. The level of estradiol( E2) in media didn’t change when cells were pretreated with the JNK inhibitor SP600125 or P38 inhibitor SB203580. Conclusion M- CSF may play an important role in promoting the proliferation and differentiation of granulosa cell,also the synthesis and secretion of estrogen.
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