Effects of vitamin D-induced supernatant of placental explants from preeclamptic women on oxidative stress and nitric oxide bioavailability in human umbilical vein endothelial cells

The study evaluated the effect of the supernatant of placental explants from preeclamptic (PE) and normotensive (NT) pregnant women after tissue treatment with or without vitamin D (VD) on oxidative stress and nitric oxide (NO) bioavailability in human umbilical vein endothelial cells (HUVEC). Placental explants were prepared from eight NT and eight PE women, and supernatants were obtained after incubation with or without hydrogen peroxide (H2O2) and/or VD. HUVEC were cultured for 24 h with supernatants, and the following parameters were analyzed in HUVEC cultures: NO, nitrate (NO3-), and nitrite (NO2-) levels, lipid peroxidation, and intracellular reactive oxygen species (ROS). Results showed that the production of NO3-, NO2-, malondialdehyde (MDA), and ROS were significantly higher in HUVEC treated with explant supernatant from PE compared to NT pregnant women, while the supernatant of PE explants treated with VD led to a decrease in these parameters. A significantly high production of NO was detected in HUVEC cultured with control supernatant of NT group, and in cultures treated with supernatant of PE explants treated with VD. Taken together, these results demonstrated that cultures of placental explants from PE women with VD treatment generated a supernatant that decreased oxidative stress and increased the bioavailability of NO in endothelial cells.

本研究评估了子痫前期（preeclamptic, PE）与血压正常（normotensive, NT）妊娠女性的胎盘外植体（placental explants），经维生素D（vitamin D, VD）或无维生素D处理后，其上清液对人脐静脉内皮细胞（human umbilical vein endothelial cells, HUVEC）中氧化应激与一氧化氮（nitric oxide, NO）生物利用度的影响。研究从8名血压正常妊娠女性与8名子痫前期妊娠女性体内制备胎盘外植体，经含或不含过氧化氢（hydrogen peroxide, H₂O₂）和/或维生素D的培养基孵育后获取上清液。将人脐静脉内皮细胞与上述上清液共培养24小时，随后检测细胞培养物中的一氧化氮、硝酸盐（nitrate, NO₃⁻）、亚硝酸盐（nitrite, NO₂⁻）水平、脂质过氧化情况以及细胞内活性氧（reactive oxygen species, ROS）。结果显示，与血压正常妊娠女性来源的外植体上清液处理组相比，子痫前期妊娠女性外植体上清液处理的人脐静脉内皮细胞中，硝酸盐、亚硝酸盐、丙二醛（malondialdehyde, MDA）与活性氧的生成量显著升高；而经维生素D处理的子痫前期外植体上清液可使上述参数水平降低。在血压正常组对照上清液处理的人脐静脉内皮细胞，以及经维生素D处理的子痫前期外植体上清液处理的细胞中，均检测到显著升高的一氧化氮生成量。综上，本研究结果表明，经维生素D处理的子痫前期妊娠女性胎盘外植体培养所得上清液，可降低内皮细胞的氧化应激水平并提升其一氧化氮生物利用度。