Huang H Y, Wen Y, Irwin J C, Kruessel J S, Soong Y K, Polan M L
Department of Gynecology and Obstetrics, Stanford University Medical Center and School of Medicine, California 94305, USA.
J Clin Endocrinol Metab. 1998 May;83(5):1721-9. doi: 10.1210/jcem.83.5.4810.
Interleukin-1 (IL-1) is expressed in human endometrium and has been shown to play an integral role in local cellular interactions during implantation. In addition, the matrix metalloproteinase (MMP) and its inhibitor, the tissue inhibitor of metalloproteinase (TIMP), are crucial during implantation, mediating in vitro trophoblast penetration, and are regulated by several cytokines expressed by trophoblast cells. We have investigated the roles of IL-1 beta and transforming growth factor-beta (TGF beta) in regulating TIMP-1, TIMP-3, and 92-kDa type IV collagenase messenger ribonucleic acid (mRNA) expression in human endometrial stromal cells using quantitative competitive PCR. Confluent stromal cell cultures treated with progesterone and estradiol for 9 days were stimulated with IL-1 beta, IL-1 beta plus anti-IL-1 beta antibody, TGF beta, and TGF beta plus anti-TGF beta antibody for an additional 24 h. Competitive complementary DNA fragments were constructed by deletion of a defined fragment from each of the target complementary DNA sequences and coamplified in quantitative competitive PCR as an internal standard. TIMP-1 and TIMP-3, but not 92-kDa type IV collagenase mRNA, were expressed in stromal cells. The 92-kDa type IV collagenase mRNA was only expressed after stimulation with IL-1 beta. IL-1 beta both augmented 92-kDa type IV collagenase mRNA expression and decreased TIMP-1 and TIMP-3 mRNA expression in a dose-dependent manner. Conversely, TGF beta augmented TIMP-1 and TIMP-3 mRNA expression, but did not affect 92-kDa type IV collagenase expression. IL-1 and TGF beta-mediated changes were both neutralized by specific antibodies. These results provide indirect evidence that IL-1 and TGF beta may play crucial roles at the embryo-maternal interface during trophoblast invasion by regulating stromal cell expression of TIMP-1, TIMP-3, and 92-kDa type IV collagenase, all of which are known to be important in trophoblast invasion.
白细胞介素-1（IL-1）在人子宫内膜中表达，并且已被证明在着床期间的局部细胞相互作用中起不可或缺的作用。此外，基质金属蛋白酶（MMP）及其抑制剂金属蛋白酶组织抑制剂（TIMP）在着床过程中至关重要，介导体外滋养层穿透，并受滋养层细胞表达的几种细胞因子调节。我们使用定量竞争PCR研究了IL-1β和转化生长因子-β（TGFβ）在调节人子宫内膜基质细胞中TIMP-1、TIMP-3和92-kDa IV型胶原酶信使核糖核酸（mRNA）表达中的作用。用孕酮和雌二醇处理9天的汇合基质细胞培养物，再用IL-1β、IL-1β加抗IL-1β抗体、TGFβ以及TGFβ加抗TGFβ抗体刺激24小时。通过从每个靶互补DNA序列中缺失一个确定的片段构建竞争性互补DNA片段，并在定量竞争PCR中作为内标共同扩增。基质细胞中表达了TIMP-1和TIMP-3，但未表达92-kDa IV型胶原酶mRNA。92-kDa IV型胶原酶mRNA仅在IL-1β刺激后表达。IL-1β以剂量依赖方式增强92-kDa IV型胶原酶mRNA表达，并降低TIMP-1和TIMP-3 mRNA表达。相反，TGFβ增强TIMP-1和TIMP-3 mRNA表达，但不影响92-kDa IV型胶原酶表达。IL-1和TGFβ介导的变化均被特异性抗体中和。这些结果提供了间接证据，表明IL-1和TGFβ可能通过调节TIMP-1、TIMP-3和92-kDa IV型胶原酶的基质细胞表达，在滋养层侵入期间的胚胎-母体界面发挥关键作用，所有这些在滋养层侵入中都很重要。
