Effects Of Simple Glycerides Comprised By Saturated Or Unsaturated Fatty Acids On The Function Of Endothelial Cells In An Atherosclerosis Model

The present investigation was undertaken to draw a comparison among the effects of Simple Glycerides, including Trilinolein (LLL), Triolein (OOO), and Tristearin (SSS), on the function of HUVEC in an AS model and to explore the potential underlying mechanisms of Simple Glycerides on impaired HUVEC. The early AS model was established by HUVEC stimulated by ox-LDL. Cell viability was measured by MTT method after the cells were treated with ox-LDL and various Simple Glycerides (including SSS,000, and LLL, respectively). Apoptosis and necrosis of the cells were determined using AnnexinV-FITC/PI staining by Flow Cytometry. The mRNAs and proteins of two well characterised endothelial ADMs, including adhesion molecule-1(ICAM-1) and endothelial cell adhesion molecule-1(E-Selectin) were respectively examined by PCR and Western Blotting. Furthermore, to discuss the different underlying mechanisms of Simple Glycerides on AS model, index of MDA and SOD in the stimulated HUVEC were determined. The main results are as follows:1. The early model of AS was established. MTT results showed that incubation of HUVEC with ox-LDL (0,50,100, and150μg/ml, respectively) for24h resulted in a concentration-dependent induction of apoptotic cell. When the concentration of ox-LDL was50μmol/L, the cell survival rate decreased to48.6%. Flow Cytometry results demonstrate that incubating with ox-LDL (50μg/ml) for24h did not significantly increase the apoptotic percentage; however,100and150μg/ml treatment resulted in an obvious increase in apoptotic cell,60.74%and72.39%, respectively. HUVEC treated with ox-LDL (50μg/ml) for24h significantly increased the percentage of apoptosis (from8.99%to15.48%) with little effect on the number of necrotic cells. Therefore,50μg/ml of ox-LDL was used to identify apoptosis.2. Cytotoxicity of Simple Glycerides on HUVEC. When the concentrations of were above100μmol/L, it showed a strong effect of Simple Glycerides on inhibiting the viability of HUVEC (p<0.05). According to the statistics, the concentrations of Simple Glycerides of no more than100μmol/L were chosen for the subsequent experiment.3. Effects of Simple Glycerides on the HUVEC in the model of AS. It is observed from the results of MTT and Flow Cytometry that LLL and OOO increased the cell viability and decreased the apoptosis percentage of ox-LDL-stimulated HUVEC. On the other hand. SSS had an adverse effect. According to the data obtained from PCR, cells simultaneously treated with LLL or000both reduced the relative expression of ICAM-1and E-selectin in EC when compared with the ox-LDL group, while SSS increased the relative expression of K AM-1and E-selectin. From the Western Blotting, the similar inclination can be observed. LLL and000may decrease the protein expression of ICAM-1and E-selectin, on the other hand, SSS have the inclination to increase these two ADM. Our observations demonstrated that LLL and SSS had the effect to prevent atherosclerosis. Besides, the anti-inflammatory effect of000was better than LLL, while SSS possesses no such beneficial effect on the development of cardiovascular disease.4. The underlying mechanism of Simple Glycerides on the HUVEC in the model of AS. This study demonstrates that both LLL and000have been implicated in the prevention and regression of atherogenesis in vitro models of AS. The improvement of antioxidant defence system and the decrease of lipid peroxidation products, MDA production were involved in the mechanism.