Molecular mechanism and functional role of macrophage colony‑stimulating factor in follicular granulosa cells

Wei,Zhang; Song,Xu; Zhifen,Zhang

doi:10.3892/mmr.2017.6857

Molecular mechanism and functional role of macrophage colony‑stimulating factor in follicular granulosa cells

Authors:
- Zhang Wei
- Xu Song
- Zhang Zhifen
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Affiliations: Department of Gynecology, Nanjing Medical University, Affiliated Hangzhou Hospital (Hangzhou First People's Hospital), Hangzhou, Zhejiang 310006, P.R. China
Published online on: June 27, 2017 https://doi.org/10.3892/mmr.2017.6857
Pages: 2875-2880

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Abstract

Our previous demonstrated that macrophage colony‑stimulating factor (M‑CSF) stimulated the production of estradiol (E2) and progesterone (P) in luteinized granulosa cells (GCs), and that its secretion may be regulated by follicle‑stimulating hormone (FSH). The present study aimed to examine the effect of M‑CSF alone or with Letrozole treatment on the function of non‑luteinizing granulosa cells, using the COV434 cell line, and its interaction with FSH. Human luteinized granulosa cells (LGC) were isolated from the follicular fluid of superovulated infertile patients (average age, 30.8±2.1 years) undergoing an intracytoplasmic sperm injection. The LGC were cultured with various concentrations of recombinant human macrophage colony stimulating factor (rhM‑CSF; 0, 10, 25, 50 or 100 ng/ml), rhM‑CSF+Letrozole (10‑6 mol/l), rhFSH (0, 10, 25, 50 or 100 IU/ml), or rhFSH+Letrozole (10‑6 mol/l). E2 concentrations in the media were measured using ELISA. The expression levels of the FSH receptor and the M‑CSF receptor were detected via reverse transcription‑quantitative polymerase chain reaction. Following COV434 cell treatment with M‑CSF, cell proliferation was quantified using the MTS assay and protein expression was detected by western blotting. It was demonstrated that M‑CSF and FSH stimulated the production of E2. The production of FSH receptors was enhanced by rhM‑CSF or rhM‑CSF+Letrozole in vitro in a dose‑dependent manner. It was observed that rhFSH promoted the expression of the M‑CSF receptor, at a certain concentration. Proliferation of COV434 cells increased in a dose‑dependent manner following treatment with RhM‑CSF. Furthermore, M‑CSF induced the phosphorylation of c‑Jun N‑terminal kinase (JNK) and p38; however, the level of E2 in the medium was not altered when the cells were pretreated with the JNK inhibitor SP600125 or the p38 inhibitor SB203580. The present study suggested that M‑CSF may be important in regulating the response of GCs to gonadotropin and may have a promotive effect in the early phase of follicular development. The biological effects of M‑CSF may partially be attributed to activation of the JNK and p38 signaling pathways. M‑CSF may represent a novel follicular development regulator agent in the future.

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1	Clark SC and Kamen R: The human hematopoietic colony-stimulating factors. J Science. 236:1229–1237. 1987. View Article : Google Scholar
2	Witt BR and Pollard JW: Colony stimulating factor-1 in human follicular fluid. J Fertil Steril. 68:259–264. 1997. View Article : Google Scholar
3	Cohen PE, Zhu L, Nishimura K and Pollard JW: Colony-stimulating factor 1 regulation of neuroendocrine pathways that control gonadal function in mice. J Endocrinology. 143:1413–1422. 2002. View Article : Google Scholar
4	Nishimura K, Tanaka N, Kawano T, Matsuura K and Okamura H: Change in macrophage colony-stimulating factor concentration in serum and follicular fluid in in-vitro fertilization and embryo transfer cycles. J Fertil Steril. 69:53–57. 1998. View Article : Google Scholar
5	Salmassi A, Mettler L, Jonat W, Buck S, Koch K and Schmutzler AG: Circulating level of macrophage colony-stimulating factor can be predictive for human in vitro fertilization outcome. J Fertil Steril. 93:116–123. 2010. View Article : Google Scholar
6	Takasaki A, Ohba T, Okamura Y, Honda R, Seki M, Tanaka N and Okamura H: Clinical use of colony-stimulating factor-1 in ovulation induction for poor responders. J Fertil Steril. 90:2287–2290. 2008. View Article : Google Scholar
7	Zhang Z, Salmassi A and Mettler L: Expression of macrophage colony-stimulating factor in human luteinized granulosa cells. Zhonghua Fu Chan Ke Za Zhi. 37:472–474. 2002.(In Chinese). PubMed/NCBI
8	Zhang Z, Salmassi A and Mettler L: Effects of macrophage colony-stimulating factor on production of estradiol and progesterone by human luteinized granulosa cells in vitro and detection of macrophage colony-stimulating factor receptor on human luteinized granulosa cells. Zhonghua Fu Chan Ke Za Zhi. 37:338–341. 2002.(In Chinese). PubMed/NCBI
9	Curry JM, Eubank TD, Roberts RD, Wang Y, Pore N, Maity A and Marsh CB: M-CSF signals through the MAPK/ERK pathway via sp1 to induce VEGF production and induces angiogenesis in vivo. J PLoS One. 3:e34052008. View Article : Google Scholar
10	Nikolic D, Calderon L, Du L and Post SR: SR-A ligand and M-CSF dynamically regulate SR-A expression and function in primary macrophage via P-38 MAPK activation. J BMC Immunol. 12:372011. View Article : Google Scholar
11	Strober W: Trypan Blue exclusion test of cell viability. Curr Protoc Immunol: Appendix. 3:Appendix 3B. 2001. View Article : Google Scholar
12	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
13	Field SL, Dasgupta T, Cummings M and Orsi NM: Cytokines in ovarian folliculogenesis, oocyte maturation and luteinisation. Mol Reprod Dev. 81:284–314. 2014. View Article : Google Scholar : PubMed/NCBI
14	Ojeda-Ojeda M, Murri M, Insenser M and Escobar-Morreale HF: Mediators of low-grade chronic inflammation in polycystic ovary syndrome (PCOS). Curr Pharm Des. 19:5775–5791. 2013. View Article : Google Scholar : PubMed/NCBI
15	Ianchiĭ RI, Voznesens'ka TIu and Shepel' OA: Cytokines and their role in reproductive system. Fiziol Zh. 53:82–90. 2007.(in Ukrainian). PubMed/NCBI
16	Luo J, Elwood F, Britschgi M, Villeda S, Zhang H, Ding Z, Zhu L, Alabsi H, Getachew R, Narasimhan R, et al: Colony-stimulating factor 1 receptor (CSF1R) signaling in injured neurons facilitates protection and survival. J Exp Med. 210:157–172. 2013. View Article : Google Scholar : PubMed/NCBI
17	Zhang Z, Fang Q and Wang J: Involvement of macrophage colony-stimulating factor (M-CSF) in the function of follicular GCs. Fertil Steril. 90:749–754. 2008. View Article : Google Scholar : PubMed/NCBI
18	Kawamura K, Chen Y, Shu Y, Cheng Y, Qiao J, Behr B, Pera RA and Hsueh AJ: Promotion of human early embryonic development and blastocyst outgrowth in vitro using autocrine/paracrine growth factors. PLoS One. 7:e493282012. View Article : Google Scholar : PubMed/NCBI
19	Araki M, Fukumatsu Y, Katabuchi H, Shultz LD, Takahashi K and Okamura H: Follicular development and ovulation in macrophage colony-stimulating factor-deficient mice homozygous for the osteopetrosis (op) mutation. Biol Reprod. 54:478–484. 1996. View Article : Google Scholar : PubMed/NCBI
20	Sweet MJ and Hume DA: CSF-1 as a regulator of macrophage activation and immune responses. Arch Immunol Ther Exp (Warsz). 51:169–177. 2003.PubMed/NCBI
21	Chitu V and Stanley ER: Colony-stimulating factor-1 in immunity and inflammation. Curr Opin Immunol. 18:39–48. 2006. View Article : Google Scholar : PubMed/NCBI
22	Schönlau F, Schlesiger C, Ehrchen J, Grabbe S, Sorg C and Sunderkötter C: Monocyte and macrophage functions in M-CSF-deficient op/op mice during experimental leishmaniasis. J Leukoc Biol. 73:564–573. 2003. View Article : Google Scholar : PubMed/NCBI
23	Zhang Z, Fang Q and Wang J: Involvement of macrophage colony-stimulating factor (M-CSF) in the function of follicular GCs. Ferti lSteril. 90:749–754. 2008. View Article : Google Scholar
24	Chakraborty P and Roy SK: Expression of FSH receptor in the hamster ovary during perinatal development. Mol Cell Endocrinol. 400:41–47. 2015. View Article : Google Scholar : PubMed/NCBI
25	Lee AW, Mao Y, Penninger JM and Yu S: Gab2 promotes colony-stimulating factor 1-regulated macrophage expansion via alternate effectors at different stages of development. Mol Cell Biol. 31:4563–4581. 2011. View Article : Google Scholar : PubMed/NCBI
26	Popa-Wagner A, Stöcker K, Balseanu AT, Rogalewski A, Diederich K, Minnerup J, Margaritescu C and Schäbitz WR: Effects of granulocyte-colony stimulating factor after stroke in aged rats. Stroke. 41:1027–1031. 2010. View Article : Google Scholar : PubMed/NCBI