3,5,3′-triiodothyronine down-regulates Fas and Fas ligand expression and suppresses caspase-3 and poly (adenosine 5′-diphosphate-ribose) polymerase cleavage and apoptosis in early placental extravillous trophoblasts in vitro
3,5,3′-triiodothyronine down-regulates Fas and Fas ligand expression and suppresses caspase-3 and poly (adenosine 5′-diphosphate-ribose) polymerase cleavage and apoptosis in early placental extravillous trophoblasts in vitro
复制标题
DOI：
10.1210/jc.2003-032208
复制
发表时间：
2004-08-01
影响因子：
5.8
通讯作者：
Maruo, T
中科院分区：
文献类型：
--
作者：
Laoag-Fernandez, JB;Matsuo, H;Maruo, T
本研究旨在探讨早期胎盘绒毛外滋养层细胞是否存在T(3)受体，以及T(3)对培养的早期胎盘绒毛外滋养层细胞Fas/Fas配体表达、caspase-3和多聚腺苷二磷酸核糖聚合酶(PARP)裂解及细胞凋亡的影响。从正常早孕胎盘组织块中分离出细胞角蛋白7和人胎盘催乳素免疫阳性的侵袭性表型胚胎干细胞，在不加或不加T(3)(10(-7)~10(-9)M)的条件下培养。免疫细胞化学、RT-PCR和Southern印迹分析检测培养的EVT中T(3)受体的存在。用激动型Fas抗体处理细胞，测定Fas敏感性。用末端脱氧核苷酸转移酶-脱氧尿嘧啶核苷缺口末端标记、流式细胞术和Hoechst核染色检测细胞凋亡。免疫细胞化学方法检测Fas、Fas配体表达及caspase-3、PARP裂解情况。早期胎盘EVT表达一个212个碱基的c-erb Abeta1转录本和T(3)受体蛋白，并在培养中表现出显著的细胞凋亡水平。T(3)可降低培养的EVT细胞Fas和Fas配体的表达，减少caspase-3和PARP的裂解，抑制细胞凋亡。虽然激动型Fas抗体的加入增加了这些细胞的凋亡，但这种反应明显被T(3)的存在所减弱。这些结果表明，T(3)受体在早期胎盘EVT中存在，T(3)通过下调Fas和Fas配体的表达来抑制细胞凋亡。这些发现与T(3)通过抑制妊娠早期细胞凋亡而促进EVT侵袭蜕膜的假说一致。
The present study was conducted to determine whether T(3) receptor exists in early placental extravillous trophoblasts (EVTs) and evaluate the influence of T(3) on Fas/Fas ligand expression, caspase-3, and poly (ADP-ribose) polymerase ( PARP) cleavage and apoptosis in cultured early placental EVTs. EVTs with invasive phenotype, isolated from normal placental explants from early pregnancy through preincubation on human fibronectin-coated dishes and exhibited cytokeratin 7 and human placental lactogen immunopositive staining, were cultured in the absence or presence of T(3) (10(-7) to 10(-9) M). The presence of T(3) receptor in cultured EVTs was examined by immunocytochemistry, RT-PCR, and Southern blot analysis. Fas sensitivity was determined by treating the cells with an agonistic Fas antibody. Apoptosis was assessed by terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling, flow cytometry, and Hoechst nuclear staining. Fas and Fas ligand expression and caspase-3 and PARP cleavage were evaluated by immunocytochemistry. Early placental EVTs expressed a 212-bp c-erb Abeta1 transcript and the T(3) receptor protein and exhibited significant levels of apoptosis in culture. Treatment with T(3) reduced the expression of Fas and Fas ligand as well as cleavage of caspase-3 and PARP and suppressed apoptosis in cultured EVTs. Although addition of agonistic Fas antibody increased apoptosis in these cells, this response was markedly attenuated by the presence of T(3). These results demonstrate that T(3) receptor is present in early placental EVTs and that T(3) suppresses apoptosis by down-regulating the expression of Fas and Fas ligand. These findings are consistent with the hypothesis that T(3) promotes EVT invasion to the decidua by suppressing apoptosis in early pregnancy.