Miller Irit, Chuderland Dana, Grossman Hadas, Ron-El Raphael, Ben-Ami Ido, Shalgi Ruth
Department of Cell and Developmental Biology (I.M., D.C., H.G., R.S.), Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv 69978, Israel; and IVF and Infertility Unit (R.R.-E., I.B.-A.), Department of Obstetrics and Gynecology, Assaf Harofeh Medical Center, Zerifin 70300, Israel.
J Clin Endocrinol Metab. 2016 Dec;101(12):4699-4709. doi: 10.1210/jc.2016-1744. Epub 2016 Sep 28.
Ovarian hyperstimulation syndrome (OHSS) is a potentially life-threatening complication of assisted reproductive technologies. This complex syndrome is known to involve massive angiogenesis and inflammation. We have previously established the anti-angiogenic involvement of pigment epithelium-derived factor (PEDF) in the pathophysiology and treatment of OHSS.
Evaluate the anti-inflammatory role of PEDF in OHSS.
In vivo mouse OHSS model and in vitro cultures of granulosa cells.
Changes in the expression of PEDF, IL-6, IL-8, and vascular endothelial growth factor (VEGF) were measured by quantitative PCR and ELISA; OHSS symptoms were recorded (body and ovarian weight gain and peritoneal vascular leakage quantified by the modified Miles's assay).
Rat granulosa cell-line stimulated with lysophosphatidic acid (LPA), exhibited a significant increase in IL-6 expression, concomitantly with a decrease in PEDF level (P < .01). Co-stimulation with recombinant PEDF (rPEDF) decreased the expression of IL-6 significantly (P < .05). Furthermore, the expression of IL-6 and IL-8 increased in LPA-stimulated human primary granulosa cells (P < .01). Co-stimulation with rPEDF decreased the expression of LPA-induced IL-6 and IL-8 mRNA and protein by 4- and 2- to 5-fold, respectively. IL-8-stimulated human primary granulosa cells exhibited increased expression of VEGF mRNA; co-stimulation with hCG induced a significantly higher increase in the expression of VEGF mRNA (P < .001), which was counteracted by rPEDF. Subcutaneous injection of 0.5 mg/kg rPEDF to OHSS-induced mice reduced the increased expression of IL-6 in the ovary (P < .01) and alleviated the severity of all OHSS parameters.
Our findings provide a framework that correlates down-regulation of OHSS symptoms caused by PEDF with both angiogenic and inflammatory pathways.
卵巢过度刺激综合征（OHSS）是辅助生殖技术中一种可能危及生命的并发症。已知这种复杂综合征涉及大量血管生成和炎症反应。我们之前已证实色素上皮衍生因子（PEDF）在OHSS的病理生理学和治疗中具有抗血管生成作用。
评估PEDF在OHSS中的抗炎作用。
体内小鼠OHSS模型及体外颗粒细胞培养。
通过定量PCR和酶联免疫吸附测定（ELISA）检测PEDF、白细胞介素-6（IL-6）、白细胞介素-8（IL-8）和血管内皮生长因子（VEGF）的表达变化；记录OHSS症状（通过改良的迈尔斯试验量化体重和卵巢重量增加以及腹膜血管渗漏情况）。
用溶血磷脂酸（LPA）刺激大鼠颗粒细胞系后，IL-6表达显著增加，同时PEDF水平降低（P < 0.01）。与重组PEDF（rPEDF）共同刺激可显著降低IL-6的表达（P < 0.05）。此外，LPA刺激的人原代颗粒细胞中IL-6和IL-8的表达增加（P < 0.01）。与rPEDF共同刺激可使LPA诱导的IL-6和IL-8 mRNA及蛋白表达分别降低4倍和2至5倍。IL-8刺激的人原代颗粒细胞中VEGF mRNA表达增加；与绒毛膜促性腺激素（hCG）共同刺激可使VEGF mRNA表达显著更高程度增加（P < 0.001），而rPEDF可抵消这一作用。对OHSS诱导的小鼠皮下注射0.5 mg/kg rPEDF可降低卵巢中IL-6的表达增加（P < 0.01），并减轻所有OHSS参数的严重程度。
我们的研究结果提供了一个框架，将PEDF引起的OHSS症状下调与血管生成和炎症途径联系起来。
