Huang H Y, Wen Y, Kruessel J S, Raga F, Soong Y K, Polan M L
Department of Gynecology and Obstetrics, Stanford University School of Medicine, Stanford, California 94305, USA.
J Clin Endocrinol Metab. 2001 Mar;86(3):1387-93. doi: 10.1210/jcem.86.3.7284.
The interleukin (IL)-1 system is a major regulator of local cellular interactions during embryonic implantation. Because IL-1beta and IL receptor antagonist (IL-1ra) are both expressed in human endometrium, we hypothesized that an appropriate ratio of IL-1beta to IL-1ra might favor the process of embryo implantation. Therefore, we investigated IL-1 regulation of the quantitative ratio of IL-1beta/IL-1ra messenger RNA (mRNA) expression in human endometrial stromal cells using quantitative competitive PCR, as well as intracellular protein expression after stromal cell solubilization. Confluent stromal cell cultures were stimulated with human IL-1beta (0-1000 IU/mL) for 24 h. After 24 h, total RNA was extracted, reverse transcribed, and coamplified by PCR with a defined amount of internal standard. The quantitative ratio was determined by the density of target to the internal standard. After culture with IL-1beta for 24 and 48 h, stromal cells were solubilized, and the intracellular protein levels of IL-1beta and IL-1ra were measured by enzyme-linked immunosorbent assay. The IL-1beta and IL-1ra mRNA were both up-regulated, and IL-1R tI mRNA was down-regulated, by IL-1beta in a dose-dependent manner. The quantitative ratio of IL-1beta to IL-1ra mRNA was constant with the presence of increasing concentrations of IL-1beta (1-1000 IU/mL). IL-1beta and IL-1ra protein was not detected in conditioned media of cultures before addition of IL-1beta. IL-1beta and IL-1ra protein levels increased with increasing amounts of IL-1beta after solubilization of stromal cells. The IL-1beta was detectable after 12 h of culture, in comparison with IL-1ra, which was detectable after 24 h of IL-1beta stimulation. These results suggest that IL-1 may play a crucial role in embryo-maternal interaction by regulating stromal cell expression of IL-1beta and IL-1ra, resulting in an appropriate ratio during the process of embryonic implantation.
白细胞介素（IL）-1系统是胚胎植入过程中局部细胞相互作用的主要调节因子。由于IL-1β和白细胞介素受体拮抗剂（IL-1ra）均在人子宫内膜中表达，我们推测IL-1β与IL-1ra的适当比例可能有利于胚胎植入过程。因此，我们使用定量竞争性PCR研究了人子宫内膜基质细胞中IL-1对IL-1β/IL-1ra信使核糖核酸（mRNA）表达定量比例的调节，以及基质细胞溶解后的细胞内蛋白表达。将汇合的基质细胞培养物用人IL-1β（0 - 1000 IU/mL）刺激24小时。24小时后，提取总RNA，逆转录，并与一定量的内标一起通过PCR进行共扩增。定量比例通过靶标与内标的密度来确定。在用IL-1β培养24小时和48小时后，溶解基质细胞，通过酶联免疫吸附测定法测量IL-1β和IL-1ra的细胞内蛋白水平。IL-1β以剂量依赖性方式上调IL-1β和IL-1ra mRNA的表达，而下调IL-1R tI mRNA的表达。随着IL-1β浓度（1 - 1000 IU/mL）的增加，IL-1β与IL-1ra mRNA的定量比例保持恒定。在添加IL-1β之前，培养物的条件培养基中未检测到IL-1β和IL-1ra蛋白。基质细胞溶解后，IL-1β和IL-1ra蛋白水平随着IL-1β量的增加而升高。与IL-1ra相比，IL-1β在培养12小时后可检测到，IL-1ra在IL-1β刺激24小时后可检测到。这些结果表明，IL-1可能通过调节基质细胞中IL-1β和IL-1ra的表达，在胚胎与母体相互作用中发挥关键作用，从而在胚胎植入过程中产生适当的比例。
