OX-LDL Induced The Overexpression Of UPP/NF-kappaB In Endothelial Cell And The Regulation Of Xuesaitong

Objective:In this study,the vitro culture of HUVEC with oxidative low density lipoprotein injury was established.To observe the effect of Xuesaitong on prevention of atherosclerosis and discuss the effect of Xuesaitong of UPP-nf-kappa B signal pathway.Method:The serum pharmacological method was used to prepare the serum:28 Male SD rats were fed adaptively for 1 week and then randomly divided into Control group(C),ox-LDL group(ox-LDL),Simvastatin group(S),low-dose Xuesaitong group(L),high-dose Xuesaitong group(H).Control group: daily distilled water(1mg/100g).Low-dose Xuesaitong group: Xuesaitong on mixed suspension of 37.5mg/kg/d.High-dose Xuesaitong group:Xuesaitong mixed suspension 75mg/kg/d.simvastatin group simvastatin mixed suspension 5mg/kg/d.Dosage regimen:one week.The last for 2 hours,chloral hydrate anesthesia,fixed in rats,the supine position,cutting the artery blood,each about 10 mL,standing for 2 hours,centrifugalling and taking supernatant to filer and set no enzyme to aseptic acid-free tube.under-20 ℃refrigerator.HUVEC cells were cultured in vitro by cell culture technology:The OX-LDLwere added to 3~6 generations of well-grown HUVEC cells for injury,and the drug containing serum was added.MTT method was used to detect OD values of 24 hours,48 hours and 72 hours respectively.The distribution of ubiquitin(Ub)in the cell status was measured by immunohistochemistry.,the expression of icam-1 and 20 S proteasomes in cells was detected by ELISA and the mRNA expression of ubiquitin activating enzyme(E1),NF-κB and Ub in the cell status was surveyed by using RealTime-PCR.Results:1.According to the analysis obtained by MTT,the optimal time of ox-ldl stimulating endothelial cell injury was at 48 hours.2.Under inverted microscope,the cells in the control group were polygonal,nuclear circle,and center,with clear boundary of cells.Ox-LDL group:decreased in the number of cells,cell gap widened,cell shrinkage,capsular shrinking and the boundary of the cell is not clear.simvastatin group:A few cell apoptosis cells was appeared and the cell morphology tended to be normal.Low dose Xuesaitong serum group:There were a few suspended apoptotic cells in the cells,and the cell morphology was normal.High dose Xuesaitong serum group:The intercellular space of the cells was narrowed and the morphology was normal.3.The expression of NF-κ B was significantly decreased and the expression of icam-1 was decreased in the Xuesaitong group.4.The expression of Ub and the expression of E1 were significantly decreased and the expression of 20 S was increased in the Xuesaitong group.5.Compared to Low-dose Xuesaitong group,the expression of NF-κBmRNA and E1 mRNA of High-dose Xuesaitong group was significantly decreased and the expression of 20 S of High-dose Xuesaitong group was increased.Conclusions:1.HUVEC were injured by ox-LDL intervention.2.The expression of NF-κB and ICAM-1 inflammatory mediators in HUVEC of ox-ldl injury can be down-regulated.3.Xuesaitong inhibits inflammation and protects vascular endothlial cells by inhibiting UPP/NF-κ B signal pathways,downing ubiquitin activating enzyme(E1)and NF-κ BmRNA expression,raising 20 s proteasome levels and inhibiting the Ub ubiquitin protein synthesis.4.The therapeutic effect of high dose serum of Xuesaitong group was better than low dose serum of Xuesaitong group in serum.