Kempisty B, Wojtanowicz-Markiewicz K, Ziółkowska A, Budna J, Ciesiółka S, Piotrowska H, Bryja A, Antosik P, Bukowska D, Wollenhaupt K, Bruska M, Brüssow K P, Nowicki M, Zabel M
Department of Histology and Embryology, Poznan University of Medical Science, Poznań, Poland.
Institute of Veterinary Sciences, Poznan University of Life Science, Poznań, Poland.
J Biol Regul Homeost Agents. 2015 Jan-Mar;29(1):39-50.
The correct functionality (sensitivity and receptivity) of endometrial tissue is regulated by paracrine and endocrine pathways that activate several mediators or metabolic pathways and gene cascades. This study aimed to investigate the influence of E2 and P4 on progesterone receptor (PGR) and progesterone receptor membrane component 1 (PGRMC1) protein expression in porcine luminal epithelial cells and their influence on the proliferation of these cells in real-time. Surface uterine luminal epithelial cells were removed using sterile surgical blades from uterine horns of ten crossbred anestrus gilts. Following treatment with collagenase I, cells were separated and transferred into 48-well E-Plates for use in a realtime cell analyzer (RTCA). The luminal epithelial cells were cultured in vitro (IVC) in standard DMEM cell culture medium and incubated with E2 (10 pg/ml, 40 pg/ml, 500 pg/ml) and P4 (10 ng/ml, 40 ng/ ml, 500 ng/ml). The cell proliferation index was analyzed after 0-240 h, 0-120 h, 120-240 h. After using the RTCA analysis we found increased proliferation of luminal epithelial cells after treatment of low doses of P4 (10 and 40 ng/ml), (P < 0.001). Higher doses of P4 led to decrease of proliferation (P < 0.001). Conversely, higher doses of E2 (500 pg/ml) increased the proliferation index as compared to low doses (10 pg/ml) and control (P < 0.001). Confocal microscopic observations revealed that higher concentrations of E2 upregulate the expression of both PGR and PGRMC1. Additionally, P4 used in lower concentrations stimulated the expression of these receptors, too. Our study presents a new influence of E2 and P4 on the expression of PGR and PGRMC1 and on the real-time proliferation of porcine luminal epithelial cells. The relationship between PGR or PGRMC1 expression and the proliferation of luminal epithelial cells may be influenced (up- or down regulated) by E2 or P4 in a steroid type- and dose-dependent manner.
子宫内膜组织的正确功能（敏感性和反应性）由旁分泌和内分泌途径调节，这些途径激活多种介质或代谢途径以及基因级联反应。本研究旨在探讨E2和P4对猪腔上皮细胞中孕激素受体（PGR）和孕激素受体膜成分1（PGRMC1）蛋白表达的影响，以及它们对这些细胞增殖的实时影响。使用无菌手术刀片从十头杂交发情期后备母猪的子宫角中取出子宫腔表面上皮细胞。用I型胶原酶处理后，分离细胞并转移到48孔E-Plates中用于实时细胞分析仪（RTCA）。将腔上皮细胞在标准DMEM细胞培养基中进行体外培养（IVC），并与E2（10 pg/ml、40 pg/ml、500 pg/ml）和P4（10 ng/ml、40 ng/ml、500 ng/ml）孵育。在0 - 240小时、0 - 120小时、120 - 240小时后分析细胞增殖指数。使用RTCA分析后，我们发现低剂量P4（10和40 ng/ml）处理后腔上皮细胞增殖增加（P < 0.001）。高剂量P4导致增殖减少（P < 0.001）。相反，与低剂量（10 pg/ml）和对照相比，高剂量E2（500 pg/ml）增加了增殖指数（P < 0.001）。共聚焦显微镜观察显示，较高浓度的E2上调了PGR和PGRMC1的表达。此外，较低浓度的P4也刺激了这些受体的表达。我们的研究揭示了E2和P4对PGR和PGRMC1表达以及猪腔上皮细胞实时增殖的新影响。PGR或PGRMC1表达与腔上皮细胞增殖之间的关系可能以类固醇类型和剂量依赖性方式受到E2或P4的影响（上调或下调）。
