The anti-atherosclerotics activity of tocotrienols (TCT) compared to alpha-Tocopherol (α-TOC) in in vitro study is not much being reported especially in human endothelial cells. The aim of the present study was to study the effects of TTMF, TCT and α-TOC on monocytes adherence to stimulated endothelial cells and to investigate the correlation between monocytes adherence and adhesion molecules in endothelial cells treated with TTMF, pure TCT isomers and α-TOC. Human umbilical vein endothelial cells (HUVECs) were incubated with TTMF, TCT isomers and α-TOC (0.3-10 µM) together with lipopolysaccharide, LPS (1 μg/ml). Monocytes adherence was measured by Rose Bengal staining. Soluble ICAM-1, VCAM-1 and e-selectin and NFκB binding were measured by ELISA. TTMF and TCT isomers inhibit monocytes adhesion to LPS-stimulated HUVECs but not α-TOC. δ-TCT exhibit the highest % inhibition of monocytes adhesion compared to the other TCT isomers. Only TCT isomers show positive correlation of monocytes adhesion with certain adhesion molecules and NFκB but not TTMF and α-TOC. In conclusion, TTMF and TCT isomers exhibit reduction of adhesion of monocytes to LPS stimulated endothelial cells. The reduction of monocytes adhesion by TCT isomers especially δ-TCT are positively correlated with reduction of adhesion molecules and NFκB deactivation. It can be suggested that TCT especially the δ-TCT isomers is beneficial in the prevention of early atherogenesis in human.
Human umbilical vein endothelial cells (HUVECs) were cultured on microcarrier beads to accommodate different experiment apparatus such as rotating wall vessel. In this study, fluid operating apparatus (FPA) was used. However, the effect of inflammation and endothelial activation biomarkers in HUVECs cultured on different culture surface and containers are not well established. The effects of temperature changes on these biomarkers in HUVECs grown in FPA, a spaceflight hardware, are still unclear. The objective of this study was to compare the protein and gene expression of inflammation and endothelial activation biomarkers in (i) HUVECs cultured on microcarrier beads in conventional culture flask (CCFMC) vs. conventional culture flask (CCF) (ii) HUVECs cultured on microcarrier in FPA (FPAMC) vs. CCFMC and (iii) HUVEC cultured in FPAMC with ideal temperature (37°C) (FPAMC) vs. simulated space travel temperature(25-37°C), (FPAMC-ST). sICAM-1 and sVCAM-1protein expression in HUVECs grown in CCFMC were higher than CCF. FPAMC had higher IL-6, TNF-α, ICAM-1, VCAM-1, e-selectin, NFκB and eNOS gene expression than in CCFMC. FPAMC-ST had higher ICAM-1 and e-selectin protein expression than FPAMC- in ideal temperature. HUVECs are cultured onto microcarrier in simulated space flight temperature compared with ideal temperature had higher protein expression of sICAM-1 and e-selectin but the protein and gene expression of other biomarkers of inflammation and endothelial activation are comparable. This suggests that differences in culture surface and container are have an impact on the expression of inflammation and adhesion molecule by HUVECs.
Cryopreservation by vitrification has been widely used in Assisted Reproductive Technology (ART) to preserve embryos for an extended period of time. However, the effect of vitrification on development of the embryos is lacking. Therefore, understanding on vitrification effects on embryonic proteins, especially those involved in preimplantation development is crucial to provide high quality embryos for further usage. In this study, XIAP and S6K1 protein expressions following vitrification was investigated, since they have been implicated in diverse cellular processes including cell growth, migration, proliferation, differentiation, survival and development of preimplantation embryos via the PI3K pathway. Embryos were obtained from superovulated female ICR mice which were mated with fertile males. The embryos were harvested at the 2-cell stage and cultured until blastocyst stage. Blastocysts were then vitrified in ESF40 cryoprotectant. Western blot was carried out to determine the expression of XIAP and S6K1 proteins. The results showed the expression of XIAP and S6K1 significantly decreased in vitrified blastocyst compared to the control. This indicates that blastocyst vitrification may impact developmental competence through the activation of apoptotic pathways.