Karwowski Wojciech, Lekesiz Katarzyna, Koc-Żórawska Ewa, Wnuczko Krzysztof, Borysewicz-Sanczyk Hanna, Naumnik Beata
Ginekol Pol. 2017;88(4):167-173. doi: 10.5603/GP.a2017.0033.
This study aims to asses the effects of estradiol vs. raloxifene on the levels of osteoprotegerin and soluble Receptor Activator of Nuclear Factor kB Ligand (sRANKL) in Human Umbilical Vein Endothelial Cells (HUVEC) culture in standard and calcifying medium.
Human Umbilical Vein Endothelial Cells were isolated from human umbilical vein by standard method. The supernatant concentrations of osteoprotegerin (OPG) and sRANKL (ELISA) were determined after incubation with glicerophosphate, estradiol , raloxifene, glicerophoshate and estradiol, glicerophosphate and raloxifene in comparison with control group at four designated time points (0, 1, 2 and 4 days of incubation).
Incubation of estradiol with HUVEC colony lowered the OPG level significantly after day 2 and 4. Meantime, the level of sRANKL was stable. Raloxifene added to standard growth medium also significantly lowered OPG concentration after day 4 only, with no impact on sRANKL concentration. When added to calcifying medium, both estradiol and raloxifene significantly changed OPG level during the experiment. In all treated groups OPG levels were lower than in groups exposed to calcifying medium only. Neither estradiol, nor raloxifene changed sRANKL levels during the experiment.
Estradiol and raloxifene affect OPG secretion from endothelial cells in vitro which may suggest their modifying role in pathogenesis of vascular calcification in postmenopausal women.
本研究旨在评估雌二醇与雷洛昔芬对人脐静脉内皮细胞(HUVEC)在标准培养基和钙化培养基中培养时骨保护素及可溶性核因子κB受体活化因子配体(sRANKL)水平的影响。
采用标准方法从人脐静脉中分离出人脐静脉内皮细胞。在与甘油磷酸、雌二醇、雷洛昔芬、甘油磷酸与雌二醇、甘油磷酸与雷洛昔芬孵育后,于四个指定时间点(孵育0、1、2和4天),通过酶联免疫吸附测定法(ELISA)测定骨保护素(OPG)和sRANKL的上清液浓度,并与对照组进行比较。
雌二醇与人脐静脉内皮细胞集落孵育后,在第2天和第4天后骨保护素水平显著降低。同时,sRANKL水平保持稳定。仅在第4天后,添加到标准生长培养基中的雷洛昔芬也显著降低了骨保护素浓度,对sRANKL浓度无影响。当添加到钙化培养基中时,雌二醇和雷洛昔芬在实验过程中均显著改变了骨保护素水平。在所有处理组中,骨保护素水平均低于仅暴露于钙化培养基的组。在实验过程中,雌二醇和雷洛昔芬均未改变sRANKL水平。
雌二醇和雷洛昔芬在体外影响内皮细胞的骨保护素分泌,这可能表明它们在绝经后女性血管钙化发病机制中具有调节作用。