| Background:With the social development and improve living standards, infectious disease cause of death are decreasing,while the atherosclerotic diseases cause the death of rapidly increasing global population has now become the top causes of death.High-calorie,high saturated fatty acid intake of too much food and lack of physical activity and other lifestyle are the culprit.Atherosclerosis(AS) are ischemic heart disease,cerebrovascular disease such as cardiovascular and cerebrovascular diseases the pathological foundation. Anti-atherosclerosis drug applications,the prevention and treatment of cardiovascular and cerebrovascular diseases has important strategic significance. Recognize human atherosclerosis is a disease and the pathogenesis of the associated research has more than 100 years of history,and the pathogenesis of AS has not yet been clear.People have made lipid infiltration theory,arterial smooth muscle cells (SMC) proliferation doctrine,thrombosis endogenous theory,response theory,such as injury hypothesis.With atherosclerosis constant depth,AS new continuous detection of cytokines,so that the inflammation has become hair happen AS an important factor in the process.In particular,inflammatory markers,high-sensitivity C-reactive protein(hs-CRP),IL-8,soluble adhesion molecule(sICAM-1), P-selectin,sVCAM-1,CD11/CD18[all at atherosclerosis have expressed and play a role.Therefore,AS clear expression of inflammatory factors in the spectrum,so as to explore the mechanism of inflammation in AS,and research into effective prevention and treatment drug of great significance.From the clinical term,there was no atherosclerosis of the concept,but according to the etiology,clinical manifestations,complications,such as the characteristics found in traditional Chinese medicine "pectoral pain with stuffiness" "apoplexy," "Vertigo," "headache," "phlegm pattern " "stasis pattern " and other records,but not just a disease.Total pathogenesis can be summarized as asthenia in origin and asthenia in superficiality,the virtual for the spleen,heart deficiency, kidney deficiency,marked as the Qi stagnation,blood stasis,and phlegm.Treatment of removing blood stasis by promoting,promoting digestion to relieve dyspepsia, promoting water metablism,reinforcing deficiency and strengthening body resistance.Chinese medicine for anti-atherosclerosis mechanisms with multi-level, multi-angle,multi-target features,mainly from the regulation of blood lipid disorders, vascular endothelial protection,inhibition of thrombosis and inhibition of vascular smooth muscle proliferation,anti-inflammatory,anti-oxidation,to improve the channels such as blood flow changes.Buyanghuanwu soup is famous in the Late Qing Dynasty by Wang Chong Ren,attending after-effects of a stroke due to blood deficiency is righteousness disadvantaged.Modern studies have proved that Buyanghuanwu soup to improve blood rheology,hypolipidemic effect, antithrombotic effect,improving myocardial ischemia,anti-atherosclerotic effect on the repair of peripheral nerve function.Qi deficiency and blood stasis and its cardiovascular and cerebrovascular disease has been of great importance to the medical profession.However,atherosclerosis is still a lack of pharmacological mechanism of the systemObjective:In this study,apolipoprotein E(ApoE) knockout mice(ApoE-/-) model of the model to study traditional Chinese medicine compound Buyanghuanwu decoction on atherosclerosis in rats,while the use of protein chip technology research atherosclerosis,as well as Buyanghuanwu Decoction inflammatory cytokines after the intervention of the expression pattern changes,further elaborated in atherosclerosis and molecular mechanism Buyanghuanwu Decoction mechanism for the clinical treatment of atherosclerosis-hardening caused by cardiovascular and cerebrovascular disease and provide a theoretical foundationMethods:24 male Apo E-/- mice were randomly divided into 3 groups of 4, respectively,for the model group(Apo E-/- + fat),Buyanghuanwu Decoction group (Apo E-/- + high Lipid + Buyanghuanwu Decoction),Zocor group(Apo E-/- + HC + simvastatin),and 8 male C57BL/6J(Apo E-/- mice derived from C57BL/6J) as a blank control group,all animals five days feeding adaptation,model group, simvastatin group,Buyanghuanwu Decoction group fed high fat diet model. Buyanghuanwu Decoction group gavage Buyanghuanwu Decoction Oral,once-daily; Zocor simvastatin group were given orally once a day for other groups,such as gavage doses of normal saline.Gavage for 14 weeks.Model after model,eyeball blood,separation of serum,-80℃refrigerator to preserve.Remove the heart and aorta,and further divided into two parts,along the inter-atrial line tip resection plane formed by the following parts,then put 10% neutral formalin solution fixed part of Add -80℃refrigerator preservedThe best use of online translation LiquiChip protein liquid system-on-chip detection of tissue factor expression profile.First of all,admission test serum add 96 filter plate,add-coupled anti-cytokine antibodies have spherical matrix(anti-1), vortex mixing at room temperature and dark shaking 2 h.And from the reaction plate with vacuum pumps at the bottom of each hole liquid removed,add detection of buffer,gently shaking again after removing the liquid,each hole and add buffer Detect biotinylated anti-two(report elements),at room temperature to avoid Light shaking 1.5 h;each hole add strept avidin-coupled phycoerythrin solution,at room temperature and dark shaking 0.5 h;add the termination of each hole buffer at room temperature after 5 min dark shaking with vacuum from the reaction plate at the bottom of each hole to remove the liquid,each machine into the hole measured 125 buffer at room temperature and darkshakinglmin;Finally LiquiChip at chip liquid detection system to read the hole on the fluorescence intensity,Part of myocardial tissue using Western and IP cell lysate protein in myocardial tissue extracts,Bradford method protein quantification,Western blot experiments carried out.Toβ-actin as a reference,the best treatment of protein samples were SDS-PAGE separation,protein electrophoresis will be transferred to the end of PVDF membrane,is closed,and incubated with two anti-anti-1,ECL color,Kodak Image Station 2000MM imaging system collected images,to obtain images using image processing software Image Tool 3.0 Determination and analysis of gray value strip to detect the expression level of p38,ERK,JNK,P-p38,P-ERK and P-JNK。Part of myocardial plasma protein extract with protein extraction kit nucleoprotein, Bradford protein quantitative method for protein gel mobility test(EMSA). Samples better deal with SDS-PAGE separation,add sample pre-electrophoresis, sample electrophoresis gel until the phenol blue reached the top 3/4 stop,and then transferred to cellulose nitrate membrane,ECL reagent add Kodak imaging workstation 2000mm images.Results 1,Effect of Buyanghuanwu decoction on atherosclerosis model of pathological changes in the effects of feeding high fat diet diet,animals were sacrificed 14 weeks after taking the heart and aorta,aortic root at the corresponding region of the slices to ensure that comparability between samples.The results showed that the model group significantly plaque formation,arterial cavity significantly smaller Buyanghuanwu Tang AS a significant reduction in plaque formation,with the model group were significantly different.2,Buyanghuanwu decoction of apoE-/- mice the effects of lipid testing four lipid found Buyanghuanwu decoction on blood lipid levels compared with the control group,no significant reduction(p＞0.05),Zocor compared with other groups to reduce significantly(p＜0.001).SHU down that it can significantly lower triglyceride(TG) and cholesterol(CHOL),increased the role of HDL.This is in rats and mice on the report is not consistent with the apoE gene deletion may be related.3,LiquiChip protein liquid system-on-chip detection of tissue factor show:with a blank group,model group,GM-CSF,IL-1,IL-6,IL-6 and TNFαwere increased significantly(p＜0.001).And compared with the model group,Group Buyanghuanwu decoction reduce TNFαexpression was significantly(p＜0.001); simvastatin group compared with the model group IL-1βand IL-6 expression reduced significantly(p＜0.001);the two compared with the model group to reduce GM-CSF and IL-10 expression increased significantly(p＜0.001).4,Buyanghuanwu decoction of MAPK family members p38,ERK and JNK protein phosphorylation and P-p38,P-ERK and the P-JNK expression.The purpose of the results by analyzing the image strip found Buyanghuanwu soup and phosphorylation of p38 protein expression compared with other groups to reduce non-significant(p＞0.05);model group ERK and JNK,P-ERK and the relative expression of P-JNK and an increase in other groups were significant(p＜0.001). Compared with model group,Buyanghuanwu decoction reduced significantly(p＜0.001).Buyanghuanwu decoction that can reduce both the level and its phosphorylation may be the regulation of cytokine expression in one of the mechanisms.5,Effect of Buyanghuanwu Decoction on NF-κB activation.Application of Pierce's Lightshift chemiluminescent EMSA Kit,by analyzing the results found that the purpose of the image strip,high fat diet can cause atherosclerosis in mice NF-κB activation,the model group compared with control group,there should be more significant(p＜0.001).And compared with the model group,and Buyanghuanwu Group Zocor reduced the difference was significant(p＜0.001).Likely to inhibit NF-κB activation reduces the transcription of some cytokines may be inhibited Buyanghuanwu Tang AS an important mechanism of inflammation.Conclusions1,Buyanghuanwu Decoction could significantly reduce atherosclerosis injury, atherosclerosis of the organizational structure with a clear protective effect.2,Buyanghuanwu decoction and Zocor both the adoption of the role of hypolipidemic reduce atherosclerosis risk factors.3,atherosclerosis,the inflammatory cytokine expression pattern of significant changes in Buyanghuanwu decoction would be a clear impact on the expression of inflammatory cytokines.4,through the protein level of authentication,Buyanghuanwu decoction significantly reduced the expression of ERK and JNK may be the regulation of cytokine expression in one of the mechanisms.5,apoE-/- mouse model of atherosclerosis in the NF-κB was activated,while the Buyanghuanwu decoction can reduce the activation of NF-κB activation of NF-κB inhibition may reduce the transcription of certain cytokines,are probably Buyanghuanwu decoction AS suppression of an important mechanism of inflammation.
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