Toth P, Li X, Lei Z M, Rao C V
Department of Obstetrics and Gynecology, University of Louisville School of Medicine, Kentucky 40292, USA.
J Clin Endocrinol Metab. 1996 Mar;81(3):1283-8. doi: 10.1210/jcem.81.3.8772613.
The present study characterized hCG/LH receptors from messenger ribonucleic acid (mRNA) to protein and whether exogenous hCG can bind and regulate the expression of the cyclooxygenase-1 (COX-1) gene in human fetal membranes from term pregnancy. Northern blotting showed that fetal membranes contain 6.0, 4.4, 2.4, and 1.4 kilobases of hCG/LH receptor mRNA transcripts. In situ hybridization revealed that amnion, chorion, and decidua contain receptor transcripts. Western immunoblotting and immunocytochemistry showed that amnion, chorion, and decidua also contain an 80-kDa receptor protein. Ligand blotting demonstrated that the 80-kDa receptor protein in fetal membranes can bind [125I]hCG, and this binding was inhibited by excess unlabeled hCG. Treatment of fetal membranes with highly purified hCG resulted in a dose- and time-dependent increase in immunoreactive COX-1 protein. The response of hCG was seen in all layers of fetal membranes. The treatment with hCG also resulted in an increase in steady state COX-1 mRNA levels. The action of hCG was prevented by cotreatment with H-89, an inhibitor of protein kinase A, but not by calphostin or lavendustin, which inhibit protein kinase C and tyrosine kinase, respectively. In summary, human fetal membranes contain hCG receptor transcripts and receptor protein that can bind hCG and up-regulate the expression of COX-1 gene.
本研究对人绒毛膜促性腺激素/促黄体生成素(hCG/LH)受体从信使核糖核酸(mRNA)到蛋白质进行了表征,并研究了外源性hCG是否能结合并调节足月妊娠人胎膜中环氧合酶-1(COX-1)基因的表达。Northern印迹分析显示,胎膜含有6.0、4.4、2.4和1.4千碱基的hCG/LH受体mRNA转录本。原位杂交显示羊膜、绒毛膜和蜕膜含有受体转录本。Western免疫印迹和免疫细胞化学显示羊膜、绒毛膜和蜕膜也含有一种80 kDa的受体蛋白。配体印迹表明,胎膜中的80 kDa受体蛋白能结合[125I]hCG,且这种结合可被过量的未标记hCG抑制。用高度纯化的hCG处理胎膜导致免疫反应性COX-1蛋白呈剂量和时间依赖性增加。在胎膜的所有层中均可见hCG的这种反应。用hCG处理还导致COX-1 mRNA稳态水平升高。hCG的作用可被蛋白激酶A抑制剂H-89共同处理所阻断,但不受分别抑制蛋白激酶C和酪氨酸激酶的钙泊三醇或拉文达ustin的影响。总之,人胎膜含有hCG受体转录本和能结合hCG并上调COX-1基因表达的受体蛋白。