O'Brien Margaret, O'Shaughnessy Deirdre, Ahamide Elodie, Morrison John J, Smith Terry J
National Centre for Biomedical and Engineering Science, Orbsen Building, National University of Ireland Galway, Galway, Ireland.
Mol Hum Reprod. 2007 Sep;13(9):655-61. doi: 10.1093/molehr/gam047. Epub 2007 Jul 30.
Extensive tissue remodelling occurs in the human myometrium before, during and after parturition. The aim of this study was to investigate the expression of two tissue remodelling molecules, matrix metalloproteinase 3 (MMP3) and alpha5 integrin (ITGA5) subunit in human myometrium, during pregnancy and labour. mRNA and protein were isolated from human pregnant labouring and non-labouring myometrial tissue, and also from human primary uterine smooth muscle cells (SMCs). Semi-quantitative RT-PCR, real-time fluorescence RT-PCR and western blotting were subsequently performed to determine the expression levels of MMP3 and ITGA5 in the myometrial tissues during pregnancy and labour, and in the primary uterine SMCs. The expression of MMP3 and ITGA5 mRNA and protein are reported for the first time during pregnancy and labour in human myometrium. Furthermore, a significant increase in expression of MMP3 mRNA (41-fold, P = 0.001), and a significant decrease in ITGA5 mRNA expression (4-fold, P < 0.001) at labour, were observed. Protein expression of these two molecules was also altered at labour, pro-MMP3 protein expression significantly increased while ITGA5 protein expression decreased, with labour onset. Expression of these molecules was also observed in primary cultured human uterine SMCs. The differential expression of these two tissue remodelling molecules at labour and their detection in uterine SMCs highlights their potential importance in myometrial function during pregnancy, labour and post-partum.
在分娩前、分娩期间和分娩后,人类子宫肌层会发生广泛的组织重塑。本研究的目的是调查两种组织重塑分子,即基质金属蛋白酶3(MMP3)和α5整合素(ITGA5)亚基在人类子宫肌层在妊娠和分娩期间的表达情况。从人类妊娠分娩期和非分娩期的子宫肌层组织以及人类原代子宫平滑肌细胞(SMC)中分离出mRNA和蛋白质。随后进行半定量逆转录聚合酶链反应(RT-PCR)、实时荧光RT-PCR和蛋白质印迹分析,以确定MMP3和ITGA5在妊娠和分娩期间的子宫肌层组织以及原代子宫SMC中的表达水平。首次报道了MMP3和ITGA5 mRNA及蛋白质在人类子宫肌层妊娠和分娩期间的表达情况。此外,观察到在分娩时MMP3 mRNA表达显著增加(41倍,P = 0.001),而ITGA5 mRNA表达显著下降(4倍,P < 0.001)。这两种分子的蛋白质表达在分娩时也发生了改变,随着分娩开始,前MMP3蛋白质表达显著增加,而ITGA5蛋白质表达下降。在原代培养的人类子宫SMC中也观察到了这些分子的表达。这两种组织重塑分子在分娩时的差异表达以及在子宫SMC中的检测突出了它们在妊娠、分娩和产后子宫肌层功能中的潜在重要性。