| 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 stable. Based on the result which TCM syndrome elements are relevant to acute cardiovascular events, we choose corresponding Chinese herbs. We further explored the optimized combination and proportion of herbal extractives in stabilizing VAP by uniform design, which is of great importance for developing effective Chinese medicine in preventing and treating AS and coronary heart disease.1 Clinical research:Multi-central research of TCM syndrome elements of hospitalized patients with coronary heart disease1.1 ObjectiveTo study distribution of TCM syndrome elements in 1005 CHD patients in hospital; to explore the correlation of TCM syndrome elements and cardiovascular events, so as to provide the clinical evidence for CHD syndrome differentiation and treatment with TCM, as well as the development of new Chinese compound prescribtion.1.2 MethodBy cross-sectional research method, we analyzed the distribution of TCM syndrome elements in 1005 CHD patients from 9 hospitals; by prospective research method, we studied the distribution of TCM syndrome elements in CHD patients with cardiovascular events in 1 year and explored the correlation of TCM syndrome elements and cardiovascular events through the principal component analysis and logistic regression analysis method.1.3 ResultThe proportion of chief TCM syndrome elements in CHD patients in descending order are blood stasis (81.4%), qi deficiency (56.8% ), turbid phlegm (48.5% ), yin deficiency (25.1%). After 1 year follow-up, there are 66 CHD patients who happen to cardiovascular events, and the other 939 CHD patients who have no cardiovascular events. The analytic results of principal component analysis and logistic regression analysis method demonstrated that the second principal component had statistical significance in logistic regression (P=0.036), and the second principal component mainly consisted of qi deficiency and yin deficiency.1.4 ConclusionBlood stasis, qi deficiency, turbid phlegm, yin deficiency were main syndrome elements of CHD patients. And that result reflected the feature of deficiency in origin and excess in superficislity. Qi deficiency and yin definciency might be the relevant TCM syndrome elements to the CHD patients who happen to cardiovascular events in 1 year follow up.2 Experimental research:Study the effect of Chinese herbs on atherosclerotic vulnerable plaque stability of ApoE gene-deficient (ApoE) mice and explor the optimized combination and proportion of chinese herbs2.1 ObjectiveBased the results of previous study that the chinses herbs of activating blood circulation and detoxicationg could stabilize atherosclerotic plaque, we combined the main sysdrome elements of CHD patients in clinic. We would study the effect of Chinese herbs on the stability of atherosclerotic vulnerable plaque in the aortic roots of fat-fed ApoE-/- mice and explore the optimized combination and proportion of Chinese herbs by uniform design.2.2 MethodWe chose giant knotweed rhizome, panax pseudo-ginseng, snakegourd fruit, and polygonum multiflorum. A uniform design with 4-factor 8-level table was used in this study. According to the table, the four Chinese herb extractive (giant knotweed rhizome, panax pseudo-ginseng, snakegourd fruit, and polygonum multiflorum) were composed for 8 medication administration groups (No.1 group-No.8 group). One hundred ApoE mice were fed a high-fat diet for 13 weeks until the mature atherosclerotic plaques formed. Thereafter, they were randomized to ten groups (10 in each group) and fed a high-fat diet for subsequent 13 weeks:8 medication administration groups, atorvastatin group, and control group. Every group was medicated with respective drug in a clinically relevant dose for another 13 weeks. Then all the mice were sacrificed at the end of experiment. For each mouse, the aorta was dissected out and stained with Sudan?for subsequent assessment of atherosclerotic lesion area. Cross sections of aortic roots were prepared and stained with H-E, a modified Movat pentachrome stain, or immunohistochemistry stain respectively. The blood was collected for determination of blood lipids. The observe items included:(1) en face whole of aorta: the atherosclerotic plaques (red stained)/ aortic intimal surface area in each aorta. (2) histological cross sectional aortic sample: the area of atherosclerotic plaque, the percentage of extracelluar lipids (EL), foam cells (FC), collagen (CA) and smooth muscle cells (SMC) in plaque. (3) Immunohistochemical staining for the expression of PPAR-?, positive staining areas were measured and expressed as a percentage of the whole plaque area. (4) Determination of serum lipid levels. The effect on plaque stability was evaluated by using the vulnerability index (?). The?was expressed as the area ratio of foam cells and extracellular lipids to collagens and smooth muscle cells. Taking the?as the screen index, we explored the optimized combination and proportion of Chinese herbs by regression analysis of uniform design.2.3 Result2.3.1 En face whole aortic sampleIn each group, the atherosclerotic plaques (red stained) were clearly observed and predominantly complex fibrous plaques, which were scattered in the entire aorta. Compared with control group, the No.2 group, No.3 group, No.6 group, No.7 group, No.8 group, and atorvastatin group was associated with a significant decrease in the plaque area (P<0.05).2.3.2 Histological cross sectional aortic sampleHE staining and Movat's pentachrome staining within the aortic cross section revealed that the atherosclerotic plaques in ApoE-deficient mice were an asymmetrically thickened intima composed with a thin fibrous cap, usually buried with foam cells, and a lipid-rich necrotic core with cholesterol clefs within the extracellular matrix, that was typical morphologic features of vulnerable plaque. In some cases, the vascular media was thinned, and elastic fibers were broken, even disappeared. The collagen was yellow stained by Movat staining, and the positive of PPAR-?expression was brown by immunohistochemical staining. As for the cross sectional area of atherosclerotic plaques within aorta, there was no significant difference among all drug treated group compared with control group (P>0.05).2.3.3 Atherosclerotic plaque compositionAs for extracellular lipids area, compared with control group, No.5 and No.7 group had a marked increase (P<0.05), the other groups and atrovastatin group had no significant difference. As for foam cell area, compared with control group, there was no significant difference among all groups (P>0.05). As for the collagen area, compared with control group, No.3 group, No.4 group, No.6 group, No.7 group, No.8 group, and atorvastatin goup was associated with a significant increase (P<0.05). As for the smooth muscle cell area, compared with control group, No.1 group, No.2 group, No.3 group, No.4 group, No.8 group, and atorvastatin group was associated with a significant increase (P>0.05).2.3.4 Expression of PPAR-?in the aortic plaqueCompared with the control group, No.2 group, No.3 group, No.4 group, No.6 group, No.7 group, No.8 group, and atorvastatin group was associated with a a marked increase in expression of PPAR-y (P<0.05).2.3.5 Plasma lipid levelsCompared with controls, there was no statistically difference among all drug treated group (P>0.05) in the level of TC, TG, LDL and HDL respectively.2.3.6 Evaluation of plaque stabilityCompared with the control group, No.1 group, No.3 group, No.4 group, No.6 group, No.7 group, No.8 group, and atorvastatin group was associated with a significant reduction in the vulnerability index (?) (P<0.05). And the vulnerability index of No.8 group was minimal (P<0.01).2.3.7 Optimized combination and proportion of Chinese herbsTaking the vulnerability index as the screen index and multivariant regression analysis of uniform design, we got the optimized combination and proportion of Chinese herbs. The first optimized combination:giant knotweed rhizome 0.3604g, snakegourd fruit 0.3604g, polygonum multiflorum 0.1834g. The second optimized combination:giant knotweed rhizome 0.3604g, polygonum multiflorum 0.1606g.2.4 ConclusionThe Chinese medicine compound prescription (giant knotweed rhizome, panax pseudo-ginseng, snakegourd fruit, and polygonum multiflorum) had a reduction in the vulnerability index (VI) in ApoE-/- mice which indicated the Chinese medicine compound prescription had potential effect on stabilizing atherosclerotic plaque. By the multivariant regression method, the optimized combination and proportion which could stabilize the atherosclerotic plaque were that the first optimized combination: giant knotweed rhizome 0.3604g, snakegourd fruit 0.3604g, polygonum multiflorum 0.1834g; the second optimized combination:giant knotweed rhizome 0.3604g, polygonum multiflorum 0.1606g, which laid a foundation of the optimized combination verificafion. |
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