A Comprehensive Investigation of the Activin-family in Chorioamnionitis

为了研究激活素在绒毛膜羊膜炎中的作用，我们首先测定了原代培养的羊膜上皮细胞中激活素A的分泌。在内部评审委员会的许可和患者的知情同意下，我们在剖宫产术中采集了单次妊娠患者的胎膜样本，这些患者没有全身疾病、早产迹象或胎儿并发症。用胰酶分散羊膜上皮细胞并进行培养。革兰氏阴性细菌细胞壁成分脂多糖和炎性细胞因子肿瘤坏死因子-α都能刺激细胞分泌激活素A。因此，激活素似乎参与了绒毛膜羊膜炎的病理过程。接下来，为了建立基因表达的综合分析系统，我们对经2，3，7，8-四氯二苯并-对二恶英(TCDD)处理的羊膜上皮细胞进行了DNA微阵列和实时定量PCR分析。两种分析方法均鉴定出39个TCDD诱导基因，其中包括典型的TCDD靶基因--CYP1A1和CYP1B1。此外，TCDD还增强了干扰素诱导基因和胶原合成或降解相关基因的表达。绒毛膜羊膜炎是胎膜早破的主要原因。另一方面，羊膜的抗张强度是由羊膜的胶原蛋白提供的。考虑到羊膜的生物学特性和目前的研究结果，本研究建立的系统可用于绒毛膜羊膜炎中包括激活素家族在内的细胞因子基因表达的全面研究。

In order to study the role of activin in chorioamnionitis, we first measured activin A secretion from the cells in primary amniotic epithelia cultures. With the permission of The Internal Review Board and the informed consent of the patients, we obtained fetal membrane samples during cesarean sections, from patients with single pregnancy without systemic disease, signs of premature delivery or fetal complications. Amniotic epithelial cells were dispersed by trypsin and cultured. Both a gram negative bacterial cell wall component lipopolysaccharide and inflammatory cytokine tumor necrosis factor-alpha stimulated the secretion of activin A from the cells. Therefore, activin seemed to be involved in the pathology of chorioamnionitis. Next, to establish a system for comprehensive analysis of gene expression, we performed DNA microarray and quantitative real-time PCR analyses in a 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) treated amniotic epithelial cell culture. Thirty nine TCDD-inducible genes, including CYP1A1 and CYP1B1 which are known as the typical TCDD target genes, was identified by both analyses. Furthermore, the expression of interferon-inducible genes and genes related to collagen synthesis or degradation was enhanced by TCDD. Chorioamnionitis is the primary cause of premature rapture of the membrane. On the other hand, the tensile strength of fetal membranes is provided by the collagen of amnion. Considering the biological character of the amnion and the present findings, the system established in the present study seems to be useful in the comprehensive investigation for gene expression of cytokines, including the activin-family, in chorioamnionitis.