Blood Detoxification Chinese Medicine Intervention In Atherosclerosis Effects And Mechanisms Of Vulnerable Plaque

Nowadays,vulnerable atherosclerotic plaque(VAP) has been highlighted for its role in leading to acute cardiovascular events.Thus,it is regarded as an important strategy to prevent the incidence of acute cardiovascular events by changing the composition of VAP to make it more stable.The theory of inflammation reaction in atherosclerosis(AS) provides innovation of pathogenesis theory and refreshment of therapeutic concept on coronary heart disease(CHD) in view of traditional Chinese medicine(TCM) with a good opportunity.Based on the results of our previous experiment study,we proposed a hypothesis that the method of activating blood circulation and detoxicating(ABCD) could stabilize atherosclerotic plaque by inhibiting inflammatory reaction,and try to confirm it in the following experimental research and clinical study.1.Experimental research:Study on the effect and underlying mechanism of herbs with function of activating blood circulation(ABC),detoxicating(D) as well as ABCD on plaque stability of ApoE gene-knockout(ApoE^-/-) mice.Objective:To observe the effects of herbal extractives with function of ABC (saponins of Panax pseudo-ginseng,SPP),D(Coptis chinensis extractive,CCE) and ABCD(giant knotweed rhizome,GKR;rhubarb extractives,RE) on the stability of VAP in the aortic roots of fat-fed ApoE^-/- mice and investigate the mechanisms in views of inhibiting inflammatory reaction,modulating lipid metabolism and improving collagen deposition and metabolism.Methods:Seventy ApoE^-/- mice were fed a high-fat diet for 13 weeks until the mature atherosclerotic plaques formed.Thereafter,they were randomized to six groups(11 in each group) and fed a high-fat diet for subsequent 13 weeks:GKR group,RE group, CCE group,SPP group,simvastatin group(positive control),and control group.Moreover, another 10 mice(C57BL/6J) was served as a normal control group.The drug-treated groups were medicated with respective drug in a clinically relevant dose for another 13 weeks.Then all the mice were sacrificed at the end of experiment.Their blood was collected for determination of blood lipids and inflammatory indices by Enzyme-Linked Immuno Srbent Assay.The morphology and composition of atherosclerotic plaques in four sections of aortic roots treated with Movat stain,HE stain and Sirius red picro-polarization method and immunohistochemistry stain respectively.The related gene expressions in the aorta of mice were determined by real-time fluorescent quantitative PCR technology.The indices includes:1) The morphology and composition of atherosclerotic plaques in aortic roots:plaque area,the percentage of extracelluar lipids,foam cells and collagen in plaque, the ratio of lipid to collagen content in plaque,the number of buried fibrous caps and the ratio of the length of fibrous cap to plaque intima.The effect on plaque stability was evaluated by using the vulnerability index(area of foam cells and extracellular lipids/area of collagens and smooth muscle cells);2) The blood lipids level and the gene expressions of perilipin and scavenger receptor CD36 associated with lipid metabolism;3 ) Inflammatory indices:serum high-sensitivity C reaction protein(hs-CRP) and soluble leukocyte differentiation Ag-40 ligand(sCD40L),the expressions of granulocyte-macrophage colony-stimulating factor(GM-CSF) and tumor necrosis factor-alpha (TNF-a),the gene expressions of peroxisome proliferator-activated receptor-γ(PPAR-γ) and nuclear factor-kappa B(NF-KB).4) Collagen metabolism:the ratio of collagenⅢtoⅠin plaque,the protein expressions of matrix metalloprotease-9(MMP-9) and tissue inhibitor-1 of metalloproteinases(TIMP-1).Results:(1) The herbal extractives with the effect of ABCD(GKR and RE) were significantly superior to ABC group(SPP) and D group(CCE) not only in improving the vulnerability index in plaque,but also in the capability of restoring fibrous cap.The comprehensive effects of the above herbs was manifested as a trend:ABCD＞D＞ABC. (2) Compared with ABC and D group,the concentrations of hsCRP and sCD40L in serum were significantly decreased in ABCD group treated with either GKR or RE,which is parallel to their effects on stabilizing atherosclerotic plaque.All of the above herbs could inhibit the expressions of GM-CSF in plaque.RE also significantly inhibited the protein expression of TNF-a,while CCE could significantly promote the mRNA and protein expression of PPAR-γ.(3) The above herbs could modulate serum lipid level to different degree and diminish extracellular lipids content in plaque.RE and CCE also remarkably inhibit the gene expression of perilipin.However,none of the above herbs showed effect on the gene expression of scavenger receptor CD36;(4) All of the above herbs could significantly increase collagen contents in plaque and reduce the ratio of collagenⅢ/Ⅰ, GKR,SPP and CCE could also reduce the ratio of MMP-9 to TIMP-1.Conclusions:The herbal extractives with function of ABC(SPP),D(CCE) and ABCD(GKR and RE) could stabilize VAP by changing plaque composition in clinically relevant doses with different mechanisms of action.The herbal extractives with function of ABCD(GKR and RE) showed superior effect on stabilizing VAP and the mechanisms of action might be related to inhibiting inflammatory reaction,modulating lipids metabolism and improving collagen deposition and metabolism.The inhibition of inflammatory reaction as the major mechanism also verified our hypothesis.2.Clinical study:The effects of the herbs with function of ABCD on the serum inflammatory markers in stable CHD patients.Objective:To observe the additional effects of Xin Qing-ning Tablets,(a representative herb with the effect of ABCD,consisting of RE),or Dan Qi Tablets(as a control with the effect of ABC,consisting of Salvia miltiorrhiza and Panax pseudo-ginseng) on the serum inflammatory markers in stable CHD patients receiving standardized statins treatment.Methods:Thirty stable CHD patients were randomized to three groups(10 in each group):the control group treated with standardized statins;the ABC group,treated with Dan Qi Tablets in addition to standardized statins treatment;the ABCD group,treated with Xin Qing-ning Tablets based on standardized statins treatment.After one month treatment, the concentrations of hs-CRP,TNF-a,MMP-9 and CD40L in serum of CHD patients before and after treatment were determined.Moreover,the blood lipids level,the scores of angina pectoris and blood stasis syndrome were also examined.Results:The ABCD group showed better effects in reducing the concentration of hs-CRP in serum,modulating blood lipid level,and improving the scores of angina pectoris and blood stasis syndrome,as compared with the control group and the ABC group.As for the concentrations of TNF-a,MMP-9 and CD40L in serum,there were no significant difference among groups.Conclusions:For stable CHD patients,the treatment with the herb with function of ABCD was superior to the treatment with function of ABC based on stabdardized statins treatment in both inhibiting inflammatory reaction and modulating blood lipids,which is consistent with our experimental results.The results also showed the better effect of ABCD group on improving the scores of angina pectoris and blood stasis syndrome as compared to ABC group.However,this is only a small-sample pilot study,whether the herbs with function of ABCD has additional benefit on preventing acute cardiovascular events still needs further demonstration with a large-scale and long-term follow-up clinical trial.