The Effects Of Simvastatin Nanoparticles On The Apoptosis Of Human Umbilical Vein Endothelial Cells Induced By Lipopolysaccharides

Objective:Preliminary prepare simvastatin nanoparticles,study its physical and chemical properties, and explore the effects of simvastatin nanoparticles on the apoptosis of human umbilical vein endothelial cells(hUVECs) induced by lipopolysaccharides(LPS).Methods:(1)Simvastatin nanoparticles were prepared by solvent diffusion method. The size of simvastatin nanoparticles were measured by Zeta potential analyzer and the drug entrapment efficiency(EE) and drug loading(DL) were determined by HighPerformance Liquid Chromatography.(2)The endothelial cells from human umbilical vein were obstained by trypsin digestion, and were cultured in vitro and identified by Immunohistochemical method.(3)Sepsis model were established with inducing human umbilical vein endothelial cells by LPS, and were divided into four group:blank control group, LPS group, simvastatin intravenous preparation (SM-IVP) group and simvastatin nanoparticles SIM-NPS) group. Cellular activity of four groups were detected by MTT method after hUVECs were co-cultured for6hours,12hours,24hours respectively. Cell apoptosis rate and the Bax protein and Bcl-2were detected by flow cytometry and western blot analysis respectively after hUVECs were co-cultured for12hours.Results:(1)The diameter of simvastatin nanoparticles were350.3±156.9nm. Entrapment efficiency (EE) and Drug loading (DL) were65.71%and3.29%respectively.(2) Primary hUVECs were spindle, paving stone shape was appeared after hUVECs were connected. The cultured cells were identified as human umbilical vein endothelial cells by Ⅷ factor related antigen antibody.(3) After hUVECs were co-cultured6hours, cellular activity of LPS group was obviously lower than that of blank control group (P<0.05), SIM-IVP group and SIM-NPS cellular activity were higher than LPS group (P<0.05), cellular activity of SIM-IVP group and SIM-NPS group has no significant difference (P>0.05). After hUVECs were co-cultured12hours and24hours, cellular activity of LPS group was obviously lower than the blank control group (P<0.05).Compared with LPS group, cellular activity of SIM-VNP group and SIM-NPS group were higher (P<0.05).SIM-NPS group had higher cellular activity than SIM-VNP group (P<0.05).There were no significant difference between two different time. After HUVECs were co-cultured12hours,the apoptosis rate of LPS group was obviously higher than the blank control group (P<0.05).Compared with LPS group, the apoptosis rate cell of SIM-IVP group and SIM-NPS group were lower (P <0.05).And SIN-NPS group had low apoptosis rate than SIM-IVP group (P<0.05). The expression of Bcl-2protein in LPS group was lower than in the blank control group (P<0.05), the expression of Bax protein in LPS group was higher than in the blank control group (P<0.05). The expression of Bcl-2proteinin in SIM-IVP group and SIM-NPS were higher than in LPS group (P<0.05), and lower than in the blank control group (P<0.05), the expression of Bax protein in SIM-IVP group and SIM-NPS was lower than in group LPS (P<0.05), and higher than in the blank control group (P<0.05). The expression of Bcl-2protein in SIM-NPS group was higher than in SIM-IVP group (P<0.05), but the expression of Bax protein in SIM-NPS group was lower than in SIM-IVP group (P<0.05).Conclusions:(1) The size of simvastatin nanoparticles which were prepared by solvent diffusion method was small, and the Entrapment efficiency and drug loadings of them were well.(2) LPS might reduce the Bcl-2protein expression, increased the Bax protein expression and finally increase cell apoptosis of hUVECs significantly.(3) SIM-IVP and SM-NPS could increased the activity of hUVECs induced by the LPS, and reduced the apoptosis rate. The protection mechanism might be that they increased the Bcl-2protein expression, and reduced the expression of Bax protein.(4) The cellular activity of SIM-NPS group was better than in SM-IVP group, and apoptosis rate was lower.It was possibly because of comparing with SM-IVP group,the expression of Bcl-2protein in SM-NPS group was higher and he expression of Bax protein in SM-NPS was lower.