Effect Of The Sumu Extract On Reverse Cholesterol Transport In Atherosclerotic Model Rats

Objective:To observe the effects of Sumu extract on reverse cholesterol transportation and its related molecular signaling pathways in vivo and in vitro,and to explore the mechanism of anti-atherosclerosis of the extract of Sumu.Methods:Experiment 1: PMA induced differentiation of human THP-1monocytes into macrophages,and incubating cells with 50?g/ml ox-LDL and0.2?Ci/ml 3H cholesterol,and applying different concentrations and different durations of Sumu extract drug-containing serum incubating cells.Oil red O staining was used to identify the morphology of foam cells;HPLC detected cholesterol levels;and scintillation counting detected cholesterol efflux rate.Experiment 2: 40 male SD rats randomly divided into control group,model group,Sumu group and simvastatin group,10 rats in each group.The control group was fed with normal diet,the other three groups were fed with high-fat diet and intraperitoneal injection of vitamin D3 for 12 weeks to establish an atherosclerosis model.In the 13 th week,the rats in the control group and the model group were given an equal volume of sodium carboxymethylcellulose solution;the Sumu group were given a suspension of Sumu extract;the simvastatin group were given simvastatin suspension.Continuous administration for 4 weeks.At the end of the 16 th week,the rats were anesthetized and weighed.The morphological changes of rat aorta and liver were observed by HE staining.The levels of serum TG,TC,LDL-C and HDL-C were measured by automatic biochemical analyzer.ELISA detected IFN-?,TNF-?,IL-6 of serum;immunohistochemistry detected CD40 L protein expression in rat abdominal aorta;Western Blot and RT-PCR detected PPAR?,LXR?,ABCA1,Cav-1,SR-BI protein and m RNA expression.Results:1 The serum containing Sumu extract promoted the cholesterol efflux of THP-1 macrophage-derived foam cells in concentration and time dependence and reduced the intracellular cholesterol content.2 HE staining show that the extract of Sumu reduced the lipid deposition in the blood vesselwall,and at the same time reduced the inflammation and swelling of the liver cells and the intracellular lipid droplets,which was similar to simvastatin.3Serum TC,TG,LDL-C and HDL-C results:Compared with the blank group,the serum TC,TG and LDL-C levels in the model group increased(P<0.01),Compared with the model group,the extract of Sumu group and simvastatin group decreased the levels of TC and TG(P<0.05,P<0.01.4Results of serum IFN-?,TNF-? and IL-6:Compared with the blank group,the serum levels of IFN-?,TNF-? and IL-6 in the model group increased(P<0.01),Compared with the model group,the extract of Sumu group and simvastatin group could decreased the levels of serum IFN-?,TNF-? and IL-6(P<0.01).5 CD40 L immunohistochemical results:Compared with the blank group,the expression intensity of CD40 L protein in abdominal aorta of model group rats increased(P<0.01).Compared with the model group,the extract of Sumu group and simvastatin group could down-regulated the expression of CD40 L protein in rat abdominal aorta(P<0.01).6Western Blot and PCR results:Compared with the blank group,PPAR?,LXR?,ABCA1,Cav-1,SR-BI protein and gene expression in liver tissue of model group rats were down-regulated(P<0.01).Compared with the model group,the extract of Sumu group and simvastatin group could up-regulated the expression of PPAR?,LXR?,ABCA1,Cav-1,SR-BI protein and gene expression in liver tissue(P<0.05,P<0.01).Conclusion:1.Drug-containing serum of the Sumu extract promotes intracellular cholesterol efflux by concentration and time-dependence;2.The extract of Sumu can alleviate the pathological damage of abdominal aorta and liver in rats with atherosclerosis;3.The extract of Sumu can reduce the levels of TC,TG,IFN-?,TNF-? and IL-6 in atherosclerotic rats,and down-regulate the expression of CD40 L protein and inhibit the inflammatory response.4.The anti-atherosclerosis effect of Sumu extract on cholesterol reversal may be partly related to the up-regulation of PPAR?,LXR?,ABCA1,Cav-1,SR-BI protein and gene expression in liver tissue.