Hum. Reprod. Advance Access originally published online on March 20, 2006
Human Reproduction 2006 21(7):1846-1855; doi:10.1093/humrep/del077
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Effects of peritoneal fluid from endometriosis patients on the release of vascular endothelial growth factor by neutrophils and monocytes
1 Department of Obstetrics and Gynecology, Pusan National University 2 Medical Research Center for Cancer Molecular Therapy, Dong-A University, Busan 3 Department of Obstetrics and Gynecology, Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University, Seoul and 4 Department of Obstetrics and Gynecology, College of Medicine, Chonnam National University, Kwang-Ju, Korea
5 To whom correspondence should be addressed at: Medical Research Center for Cancer Molecular Therapy, Dong-A University, Busan 602-714, Korea. E-mail: jykwak{at}dau.ac.kr
6 To whom correspondence should be addressed at: Department of Obstetrics and Gynecology, Medical Research Institute, Pusan National University, Busan 602-739, Korea. E-mail: kuslee{at}pusan.ac.kr
BACKGROUND: An increase in the level of the vascular endothelial growth factor (VEGF) production has been reported in the peritoneal fluid (PF) of endometriosis patients. This suggests that changes in the vascular permeability and angiogenesis play an important role in the pathophysiology of this disease. This study examined the effects of the PF obtained from endometriosis patients on the release of VEGF by neutrophils and monocytes. METHODS: Neutrophils and monocytes were obtained from young healthy volunteers and cultured with the PF obtained from either endometriosis patients (EPF) (n = 18) or a control group (CPF) (n = 4). A human monocyte/macrophage cell line, THP-1, was cultured with either 10% EPF or 10% CPF. The PF and culture supernatants were assayed for VEGF using ELISA. Real-time PCR and Western blotting were used to measure the VEGF mRNA and protein expression level, respectively. RESULTS: The VEGF levels were higher in the EPF than in the CPF (591 ± 75 versus 185 ± 31 pg/ml, P < 0.05). However, the level of VEGF released by THP-1 cells in CPF and EPF was similar. The EPF induced the release of VEGF by neutrophils, but no VEGF was released by monocytes. The VEGF mRNA expression levels in the neutrophils were higher in the EPF, which was abrogated by cycloheximide, suggesting that the EPF induces the production of VEGF in neutrophils. Neutralizing antibodies against IL-8 and TNF-
did not completely prevent the EPF-induced release of VEGF by the neutrophils, even though these growth factors stimulated the release of VEGF by neutrophils. There was a positive correlation between the VEGF and IL-10 concentrations in the EPF (correlation coefficient = 0.549, P = 0.012, n = 18), but the neutralizing antibody of IL-10 did not affect the release of VEGF by the EPF-treated neutrophils. CONCLUSION: The EPF induced the production and release of VEGF by neutrophils, suggesting that neutrophils may be a source of peritoneal VEGF. In addition, neutrophil-derived VEGF might be a marker for diagnosing endometriosis.
Key words: endometriosis/monocytes/neutrophils/peritoneal fluid/VEGF
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