Effect Of Soufeng Fu - Shu Chinese Medicine On AS Unstable Plaque In ApoE Gene Knockout Mice

Purpose： In this study, through the establishment of ApoE knockout mice [ApoE(-/-) mice] atherosclerosis model, using immunohistochemistry and molecularbiology techniques to detect heme oxygenase-1(HO-1) and heat70(HSP70) shockprotein expression levels to explore atherosclerotic plaque instability andinflammation factors in the relationship, in order to clarify the law found thewind expectorant herbs on ApoE knockout mice unstable atherosclerotic plaqueintervention mechanisms to provide a scientific basis for the prevention andtreatment of atherosclerosis medicine.Material and method：1.6to8-week-old ApoE (-/-) mice were70high fat diet for13weeks weresacrificed4, the abdominal aortic root, HE staining AS sclerosis foundationdegree, determine the successful model will be made successful model of ApoE(-/-) mice60were divided into five groups each of12, that in the model group,stable plaque soup low dose group, middle dose group and high dose group andwestern medicine group, continue to give high-fat diet feeding; normal mice12to blank control group given normal diet. Blank control group and model groupwere given saline0.3mL/day; according to the recommended adult dailymedication commonly used in clinical dose, body surface area of adult miceconversion coefficient0.0026and converted into the amount of mouse: stablelow dose group6.175soup spots g/kg/day dose group12.35g/kg/day, high-dosegroup24.7g/kg/day, western medicine group (atorvastatin calcium)27.3mg/kg/day. Gavage once a day. After13weeks of administration, the abdominal aorta,pathological observation.2．Would remove the ApoE-mouse aortic tissue using real-time quantitative RT-PCRtechnology to detect and HE staining and immunohistochemical staining using light microscopy abdominal aorta compared six groups of HSP70and HO-1theexpression levels to explore atherosclerotic plaque instability andinflammation factors in relationships.Results：1．ApoE (-/-) Mice abdominal aortic HE staining results in the light microscopeControl group: the arterial intima with a layer of endothelial cells,endothelial cell layer exhibits folds like arrangement, but the arrangementuniform, smooth surface, no foam cells and inflammatory cells.Model group: Obvious endovascular atherosclerotic plaques appear, the plaquesurface with a fibrous cap, mainly composed of fibrous cap of smooth muscle cells,elastic fibers and collagen tissue, seen a lot of the central plaque lipidaccumulation, plaque seen in a large number of foam cells, and occasionallyinflammatory cell infiltration. Intimal arterial plaque area appear seriouslydamaged.Stable plaque soup low-dose group: Endovascular visible atheroscleroticplaque, plaque area was significantly smaller than the model group, the plaquelipid content less than the model group, the plaque shows a small amount of foamcells and inflammatory cells. Occasionally endometrial incomplete.Stable plaque soup, high-dose group, western medicine group: Most of thearterial intima more smooth, occasionally endometrial insufficiency,endometrial area can be seen clearly incomplete block the formation of fattystreaks, occasionally foam cell aggregation, a small amount of cross-fibercomponents Union.2.HO-1protein and HSP-70expression results(1) HO-1protein expression results: Compared with the control group, modelgroup HO-1protein expression was significantly lower (P <0.01). Compared withthe model group, stable plaque soup each dose group, western medicine groupApoE (-/-) was significantly elevated levels of HO-1protein expression inmice abdominal aortic tissue (P <0.01), stable plaques soup, high doses no significant difference (P>0.05) between the group and western medicine group.HO-1protein expression positive cells in the vascular endothelial cells (EC),foam cells and vascular smooth muscle cells (VSMC) inside, brown granulescontaining cells of HO-1protein expression. The positive control group cellswere mainly distributed in the intima, HO-1protein expression in the EC; modelgroup HO-1protein expression was significantly reduced, occasionally weakexpression; stable plaque soup each dose group and western medicine group wassignificantly higher expression levels, is widely expressed in the EC, VSMCfoam cells and plaque. The results suggest that: steady spot soup each dosegroup could increase the expression of HO-1protein levels, which in the high-dose group and atorvastatin calcium effect similar.(2) HSP-70protein expression showed: Compared with the control group, modelgroup HSP-70protein expression was significantly lower (P <0.01); compared withthe model group, stable plaque soup, the high-dose group, western medicinegroup ApoE (-/-) was significantly higher (P <0.05or P <0.01) in mice abdominalaortic tissue protein levels of HSP-70, and no significant difference (P betweenstable plaque soup high-dose group and western medicine group>0.05). HSP70is expressed mainly in the cytoplasm, the positive expression products werebrown particles of atherosclerotic plaque fibrous cap mainly expressed in thecytoplasm of smooth muscle cells. The control group of mice within the arterialintima seen a lot of positive expression of cytoplasmic granules, model miceartery biopsy material was significantly reduced in expression, the expressionlevels of the treatment groups than the model group. The results suggest that:steady spot soup, high-dose group could raise the level of HSP-70proteinexpression, in which the high-dose group and atorvastatin calcium effectsimilar.3.HO-1mRNA expression and HSP-70’s results(1) HO-1mRNA expression showed that: Compared with the control group,-1HO mRNAexpression in model group was significantly lower (P <0.05); compared with themodel group, stable plaque soup each dose group, western medicine group ApoE (-/-) was significantly higher (P <0.01), no significant difference (P betweenstable plaque soup high-dose group and western medicine group> mouse aorta tissueHO-1mRNA expression level of0.05). The results suggest that: steady spot soupeach dose group could increase HO-1mRNA expression levels, including high-dosegroup and atorvastatin calcium similar role.(2) HSP-70mRNA expression showed that: Compared with the control group, modelgroup HSP-70mRNA expression was significantly lower (P <0.01); compared withthe model group, stable plaque soup each dose group, western medicine group ApoE(-/-) was significantly higher (P <0.01), no significant difference (P betweenstable plaque soup high-dose group and western medicine group> mouse aorta tissueHSP-70mRNA expression level of0.05). The results suggest that: steady spotsoup each dose group could increase HSP-70mRNA expression levels, includinghigh-dose group and atorvastatin calcium similar role.Conclusion：1. ApoE knockout mouse model of atherosclerosis success.2.2. The experimental model group, HO-1, HSP70expression levels weresignificantly reduced.3.Stable plaque soup can improve ApoE (-/-) mice abdominal aortic tissue HO-1,the expression levels of HSP70, to inhibit the development of atherosclerosisby inhibiting inflammation and regulate immune responses, and thus play astabilizing plaque effect.