| Atherosclerosis(AS)is the common pathological basis of cardiovascular and cerebrovascular diseases such as hyperlipidemia,coronary heart disease,and hypertension.It is mainly manifested as vascular endothelial damage,cell adhesion molecules,and cell adhesion molecules.The expression of CAMs was up-regulated and lipids were infiltrated,macrophages and smooth muscle cells were accumulated in the blood vessels,the elasticity of the blood vessel wall was lost,and atherosclerotic plaques were formed,involving the middle and large arteries.The number of people who die from AS related diseases is up to 20 million every year,which threatens the life and health of all mankind.There is no exact disease name of atherosclerosis recorded in traditional Chinese medicine.According to the specific clinical manifestations,atherosclerosis can be classified into "chest stuffiness","stroke","heartache","pulse volume" and so on.Due to inherent deficiency or acquired malnutrition,the five viscera are damaged,and the spleen and stomach have dysfunction in transportation of water,grain,and body fluid,producing pathological products such as phlegm and blood stasis,which are prone to gather into clumps and block the veins,causing adverse blood circulation and presenting as conglomeration.Traditional Chinese medicine believes that blood stasis runs through the onset of AS,and the method of activating blood circulation and dispersing stasis is often used to treat it.Danggui Shaoyao powder is a classic prescription of Jingui Yaolue,which has the functions of activating blood and disperses blood stasis,nourishingblood and blood circulation.It is in line with the syndrome characteristics of AS with phlegm turbidity endogenous,phlegm and blood stasis mutual accumulation in the vein.Based on the theory of"correspondence between prescription and syndrome elements",this study separated the elements of "Huoxue Sanyu" from Danggui Shaoyao powder.Taking platelet aggregation rate and vascular endothelial adhesion factor AS the entry points,this study used network pharmacology and molecular docking technology to explore the anti-AS targets of Danggui Chuanxiong Rhizoma,in order to provide reference for clinical practice.Objective(1)Pharmacodynamic study of Danggui Shaoyao powder and "Huoxue dispersing stasis"in the prevention and treatment of AS in mice.(2)To explore the effects of Danggui Shaoyao powder and "Huoxue dispersing stasis"formula elements on AS mice by network pharmacology combined with in vivo animal experiments,and to explore the possible corresponding targets.MethodsThirty-six ApoE-/-mice were fed with high-fat diet and randomly divided into model group,Danggui Shaoyao powder group and Huoxue dispersing powder stasis group,with 12 mice in each group.Twelve normal C57BL/6J mice were used as the normal group and fed with normal diet.After 16 weeks of continuous intervention,the general condition of the mice was recorded,the aorta of the mice stripped,and the aorta was stained with improved oil red O staining,etc.Image-pro plus 6.0 Image processing software was used to calculate the percentage of aortic plaque area.Paraffin sections were stained with HE staining to observe the pathological changes.The dissected aorta was stained with HE.Platelet aggregation rate was detected by turbidimetry,and the efficacy of Huoxue Sanyu was evaluated.Immunohistochemistry was used to detect intercellular adhesion molecule-1(ICAM-1)and vascular cell adhesion molecule-1(VCAM-1)in mouse aortic vascular endothelial cells.RT-qPCR was used to measure the mRNA expression of VEGFR-2,PTGS2 and PGI2 in vascular tissues of mice in each group.Results(1)Body weight Compared with the normal group,the body weight of the model group increased;Compared with the model group,the body weight of the Danggui Shaoyao powder group and Huoxue dispersing stasis group decreased.At the same time,by comparing the body weight of the mice at the 16th week with the initial body weight,it was found that the growth value of the mice in the model group was the largest(P<0.01 or P<0.05).(2)Oil red O staining of aorta There was no obvious red staining area in the normal group,indicating no atherosclerotic plaque formation.Compared with the normal group,the model group had a significant increase in the red colored area of the aorta(P<0.05),suggesting the formation of atherosclerotic plaques in the aorta of the model group.Compared with the model group,the Danggui Shaoyao powder group and Huoxue dispersing stasis group had significantly reduced red colored areas(P<0.05).(3)Liver histopathological observation Compared with the model group,the structure of liver cells in the Danggui Shaoyao powder group and Huoxue powder group was normal,the liver cords were arranged neatly,and there was no obvious lipid droplet vacuoles in the central vein cells.There was no significant difference in liver HE staining between the groups.(4)HE staining of aorta The structure of the inner,middle and outer membranes of the aorta in the normal group was clear and arranged neatly,and there was no plaque in the arterial wall.In the model group,the vascular intima was significantly thickened,the endothelial cells were arranged in disorder,there were obvious plaques protruding into the lumen,and a large number of lipid deposits,showing typical pathological changes of AS.Compared with the model group,the thickness of aortic plaque in the Danggui Shaoyao powder group and Huoxue dispersing stasis group was reduced.(5)Platelet aggregation rate Compared with the normal group,the platelet aggregation rate in the model group was significantly increased(P<0.01).Compared with the model group,the platelet aggregation rate was significantly decreased in the Danggui Shaoyao powder group and Huoxue dispersing stasis group(P<0.01).(6)Evaluation of curative effect Compared with the normal group,the evaluation of curative effect of the model group was significantly reduced(P<0.01).Compared with the model group,the efficacy evaluation of Danggui Shaoyao powder group and Huoxue Powder stasis group was significantly increased(P<0.01).(7)Vascular endothelial adhesion factor Compared with the normal group,the expression of ICAM-1 and VCAM-1 in the aorta of the model group was significantly increased(P<0.01).Compared with the model group,the expression levels of ICAM-1 and VCAM-1 in aorta of the Danggui Shaoyao powder group and Huoxue dispersing stasis group were significantly decreased(P<0.01).(8)The mRNA expression of VEGFR-2,PTGS2 and PGI2 Compared with the normal group,the expression of VEGFR-2 and PTGS2 in the model group was increased,and PGI2 was significantly decreased(P<0.01).Compared with the model group,the expression of VEGFR-2 and PTGS2 genes in the Danggui Shaoyao powder group and Huoxue dispersing stasis group decreased,and PGI2 increased significantly(P<0.01 or P<0.05).Conclusion(1)Danggui Shaoyao powder and its "Huoxue decoction and stasis" formula can improve the weight gain of AS mice induced by high-fat diet,improve the oil red O staining of aorta,reduce the platelet aggregation rate,down-regulate the expression of endothelial adhesion factor,and reduce the aortic plaque area,thereby intervening the process of AS.(2)Network pharmacology analysis suggested that Danggui sinensis and Chuanxiong Rhizoma,the ingredients of "Huoxue Sanyu" formula,exerted anti-AS effects through multiple components,targets,biological processes and signaling pathways.Danggui sinensis and Chuanxiong may act on KDR,MAPK14,CASP9,PGI2,PTGS2,NOS3 and other targets through stigmasterol,sitosterol,β-sitosterol,cyclodextrin and other active ingredients.By regulating VEGF signaling pathway,PPAR signaling pathway,AMPK signaling pathway,it plays a role in improving blood lipids,regulating vascular endothelial function.(3)Animal experiments showed that Danggui Shaoyao powder and Chuanxiong Rhizoma could inhibit the high expression of VEGFR-2 and PTGS2 genes in mice,and promote the expression of PGI2,thereby interfering with the development of AS.This study provides experimental basis for studying the mechanism of action of Danggui Shaoyao powder and its prescription elements of "Huoxue Decoction and stasis",and provides reference for further research on its anti-AS effect. |
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