Modulation Of Selenium On Angiogenesis Of HUEVC, Activity Of Angiotension Converting Enzyme And Macrophage Up-taking Of Low Density Lipoprotein

Effects and molecular mechanisms of SePhen on angiogenesis in vitro of human umbilical vein endothelial cells (HUEVC) were investigated. The functional inhibition of selenium (Se) containing peptides derived from Se-enriched Spirulina platensis (SeSP-Ps) to angiotensin convert enzyme (ACE) was explored. The impacts of low, medium and high Se status on lipids up-taking of bone marrow derived macrophages (BMDM) were also studied. The experimental design and main results were shown as following:1. The HUVEC cultures were treated with SePhen at4～6μM for12h. The cells proliferation was detected by MTT assay. The cells migration was measured by in vitro scratch assay, and the angiogenesis of HUVEC was determined by in vitro angiogenesis assay on the extracellular matrix gel at the indicated dosage and time point. We found that the cells proliferation, migration and angiogenesis of HUEVC were decreased by SePhen treatments with a dose-dependent manner.2. SeSP-Ps was prepared by the hydrolysis process of combination of pepsin-trypsin and chimotrypsin, and then ACE activity dynamic assay was performed to show functional inhibition of SeSP-Ps. We found the in vitro IC50of SeSP-Ps to ACE activity was74.6μg/mL.3. Mice models with low, medium and high Se status were made by feeding standard controlled Se diets during growth of4-12weeks. BMDMs were isolated and induced from the different Se status mice, and then were cultured with80,160, and320ng/mL of Na2SeO3/Se(Ⅳ), respectively. Uptake of oxidized LDL in BMDM was measured by Dil-fluorescent probe labeled oxidized LDL (Dil-Ox-LDL) after5h incubation followed with or without LPS priming for6h. Activities of GPx1, GPx4and Trxrd were assayed by Kits. Expression levels of GPx1, GPx4, Trxrd, CD36and PPARy were detected by Western blot and/or real time PCR (RT-PCR). ROS was monitored by flow cytometry assay. We found that activities and protein expression levels of selenoenzymes of GPx1, GPx4, and Trxrd were up-regulated with increasing Se status. We also found that uptake of LDL was decreased in BMDMs with or without LPS priming. Interestingly, uptake of LDL induced significant lower levels of CD36protein in M-and H-Se than L-Se BMDMs. In addition, ROS generation of M-and H-Se BMDMs was significant higher than that of L-Se BMDMs with LDL treatments.In conclusion, our present data suggest that SePhen might be a potential antagonistic drug for angiogenesis of HUVECs, the SeSP-Ps has functional inhibition to ACE, and adequate Se status plays important roles in uptake of LDL in macrophages through regulation degradation of CD36and ROS generation.