Huber A V, Saleh L, Prast J, Haslinger P, Knöfler Martin
Department of Obstetrics and Gynecology, Medical University of Vienna, AKH, Waehringer Guertel 18-20, A-1090 Vienna, Austria.
Mol Hum Reprod. 2007 Aug;13(8):595-604. doi: 10.1093/molehr/gam032. Epub 2007 May 24.
Recently, a clinical study provided evidence that treatment of endometriotic women with human chorionic gonadotrophin (hCG) alleviates disease-related pain and sleeplessness suggesting therapeutic effects of the hormone. Since endometriosis is associated with aberrant concentrations of inflammatory mediators in the peritoneal fluid, we investigated whether hCG may affect cytokine-dependent activation of the key-regulatory transcription factor NF-kappaB and expression of two nuclear factor kappa B (NF-kappaB)-inducible genes, tumour necrosing factor (TNF-alpha) and interleukin (IL)-1beta, in stromal cells isolated from ectopic endometriotic tissues. Electrophoretic mobility shift assay revealed that treatment of these cultures with the urinary preparation hCG-A suppressed TNF-alpha- or IL-1beta-induced NF-kappaB DNA-binding activity, whereas another urinary hCG preparation (hCG-B) was less effective. Recombinant alphahCG or epidermal growth factor (EGF), a contaminant of some urinary hCG preparations, did not alter cytokine-dependent NF-kappaB activation. Immunofluorescene of its p65 subunit revealed that pre-incubation with hCG-A strongly decreased TNF-alpha-dependent nuclear expression of NF-kappaB. Accordingly, hCG-A diminished IL-1beta-induced TNF-alpha transcript levels and protein release measured by quantitative real-time PCR and enzyme-linked immunosorbent assay. The hormone also attenuated TNF-alpha-dependent mRNA expression of IL-1beta. Western blot analyses revealed that hCG-A impaired TNF-alpha-mediated phosphorylation and degradation of the inhibitor IkappaBalpha suggesting that the hormone may reduce nuclear import of NF-kappaB by stabilizing its inhibitor. The data suggest that hCG attenuates inflammation-dependent NF-kappaB activation and cytokine expression that could provide one explanation for the beneficial role of the hormone in endometriotic patients.
最近，一项临床研究提供了证据，表明用人绒毛膜促性腺激素（hCG）治疗子宫内膜异位症女性可减轻疾病相关的疼痛和失眠，提示该激素具有治疗作用。由于子宫内膜异位症与腹膜液中炎症介质浓度异常有关，我们研究了hCG是否可能影响异位子宫内膜组织分离的基质细胞中关键调节转录因子核因子κB（NF-κB）的细胞因子依赖性激活以及两个核因子κB（NF-κB）诱导基因肿瘤坏死因子（TNF-α）和白细胞介素（IL）-1β的表达。电泳迁移率变动分析显示，用尿制剂hCG-A处理这些培养物可抑制TNF-α或IL-1β诱导的NF-κB DNA结合活性，而另一种尿hCG制剂（hCG-B）效果较差。重组α-hCG或表皮生长因子（EGF，某些尿hCG制剂中的污染物）不会改变细胞因子依赖性NF-κB激活。对其p65亚基的免疫荧光显示，与hCG-A预孵育可强烈降低TNF-α依赖性NF-κB的核表达。相应地，hCG-A通过定量实时PCR和酶联免疫吸附测定法降低了IL-1β诱导的TNF-α转录水平和蛋白质释放。该激素还减弱了TNF-α依赖性IL-1β的mRNA表达。蛋白质印迹分析显示，hCG-A损害了TNF-α介导的抑制剂IκBα的磷酸化和降解，提示该激素可能通过稳定其抑制剂来减少NF-κB的核转运。数据表明，hCG减弱炎症依赖性NF-κB激活和细胞因子表达，这可能为该激素在子宫内膜异位症患者中的有益作用提供一种解释。
