Compatibility Rule Of Traditional Chinese Medicine In The Treatment Of Carotid Atherosclerosis And Study On Plaque Stabilization By Ligustrazine

Atherosclerosis is a disease in which the wall of the artery develops lesions,composed of large amounts of lipids and necrotic debris.In addition to coronary arteries,carotid arteries are also a common lesion of atherosclerosis,which can reflect the severity of atherosclerosis progression.Previous studies have shown that carotid artery plaques significantly increase the risk of stroke and death.China bears the biggest burden of stroke worldwide,with significantly high incidence and mortality rate of ischemic stroke.Traditional Chinese medicine,as a unique treatment with Chinese characteristics,plays a crucial role in the treatment of cardiovascular and cerebrovascular diseases,especially in the field of therapeutics development to activate blood circulation and remove blood stasis,and has achieved relatively rich research results.However,no systematic summary has been found on the prescription rules of traditional Chinese medicine in the treatment of carotid atherosclerosis.In addition,it is necessary to study the mechanism of herbal compounds inhibiting atherosclerosis due to their effects on activating blood circulation and removing blood stasis.Therefore,the following two aspects were studied in this thesis:(1)Data mining of traditional Chinese medicine prescriptions for carotid atherosclerosis was carried out by using traditional Chinese medicine inheritance support system;(2)LDLR-/-Golden Hamsters were used to investigate the effect and mechanism of ligustrazine,a natural compound extracted from traditional Chinese herbs for promoting blood circulation and removing blood stasis,thereby inhibiting the development of atherosclerosis.Part 1 Study on the rule of prescription for carotid atherosclerosis via traditional Chinese medicine inheritance support systemObjective:This study was designed to explore the rules of the prescription of traditional Chinese medicine for the treatment of carotid atherosclerosis.Methods:The keywords of "carotid atherosclerosis","carotid stenosis","carotid occlusion","atherosclerosis","carotid artery","traditional Chinese medicine","Chinese medicine","TCM therapy","random",and "traditional Chinese medicine" were searched in CNKI,VIP and Wanfang database.Randomized controlled studies of carotid atherosclerosis patients treated with Chinese herbal compound were included.Data were extracted independently by two researchers,and the traditional Chinese medicine inheritance support system was used to analyze the prescriptions,including the analysis of four qi and five flavours,drug pairs and new prescriptions.The results were presented in tables or network visualization.Results:Among the traditional Chinese medicine prescriptions for the treatment of carotid atherosclerosis,the common medicinal properties were warm drugs,cold drugs and flat drugs.Most of the medicines were attributed to sweet taste,bitter taste and acid taste,and most of the drugs were attributed to liver meridian,spleen meridian and heart meridian.The drugs used frequently were Danshen,Chuanxiong and Huangqi.The drug pairs included "Chuanxiong,Danshen","Danshen,Huangqi","Chuanxiong,Huangqi","Danshen,Danggui" and "Banxia,Danshen".In addition,the key drug paris were as follows:"Chenpi,Gancao,Fuling ","Quanxie,Wugong,Tubiechong ","Heshouwu,Juemingzi,Shanzha","Bingpian,Zhebeimu,Xiebai"," Dangggui,Chuanxiong,Chishao,Honghua"," Quanxie,Wugong,Yimucao","Shanzha,Heye,Jiaogulan","Bingpian,Renshen,Shexiang"."Chenpi,Gancao,Fuling,Dangggui,Chuanxiong,Chishao,Honghua" can activate blood circulation and remove blood stasis,regulate qi and dissolve phlegm."Quanxie,Wugong,Tubiechong,Jiangcan,Yimucao"can activate blood circulation,remove blood stasis and eliminate the lumps;"Heshouwu,Juemingzi,Shanzha,Heye,Jiaogulan" is specially designed to help digestion,and reduce lipid,which is consistent with the emphasis on the control of blood lipid in the medical treatment of Western medicine."Bingpian,Zhebeimu,Xiebai,Renshen and Shexiang" can nourish qi,strengthen the heart and reduce phlegm.Conclusion:The drugs for the treatment of carotid atherosclerosis included Danshen,Chuanxiong,and Huangqi.The drug pairs included "Chuanxiong,Danshen","Danshen,Huangqi","Chuanxiong,Huangqi","Chenpi,Banxia" and "Fuling,Banxia".The principle of the treatment is to activate blood circulation,remove blood stasis,replenish qi and remove phlegm,with methods of reducing blood lipid.Part 2 The study on the effect of ligustrazine on inhibiting platelet activation and stabilizing atherosclerotic plaquesObjective:To observe the effects and the underlying mechanism of traditional Chinese medicine ingredient(ligustrazine)on blood lipid level,plaque area and composition,platelet activation by using golden hamster models.Methods:LDLR-/-golden hamsters were fed with high-fat and high-cholesterol diet.The animals were given distilled water(vehicle)and clopidogrel,ligustrazine via oral gavage.Then the total cholesterol(TC),triglyceride(TG),high-density lipoprotein(HDL)and non-high density lipoprotein(non-HDL)levels were determined.The progression of aortic atherosclerotic plaques were overserved using frozen section via Oil red O,CD68,α-SMA staining.En face analysis was performed using aortas.I isolated the washed platelets after adenosine diphosphate(ADP)activation.Then the analysis of intracellular adenosine cyclophosphate(cAMP),and calcium concentration were determined.Western blot was chosen to detect the expression levels of phosphatidyl inositol 3 kinase(PI3K),serine/threonine protein kinase(Akt),the phosphorylated Akt(Thr308),and the phosphorylated Akt(Ser473)proteins.Results:After 8 weeks of high-fat and high-cholesterol diet,hamsters developed severe hyperlipidemia and had atherosclerotic plaques in the aortic root and the whole aortas.Clopidogrel had no significant effect on blood lipid levels.The lipid analysis showed TC and non-HDL in ligustrazine group were significantly decreased than that of model group(P<0.05).After 8 weeks of administration of drugs,the Oil red O staining of frozen sections of aortic root in model group showed obvious atherosclerotic plaques.Clopidogrel(P<0.05)and ligustrazine group(P<0.01)could significantly reduce the plaque area.In addition to lipid deposition,the infiltration of macrophages and the reduction of the number of smooth muscle cells are also pathological manifestations of decreased plaque stability.Compared with the wild type(WT)group(P<0.01),macrophage infiltration was observed in the sections of the model group.Both clopidogrel(P<0.05)and ligustrazine group(P<0.05)could inhibit the macrophage infiltration.Compared with the WT group(P<0.05),the number of smooth muscle cells decreased significantly in the sections of the model group,and the relative number of smooth muscle cells in clopidogrel(P<0.05)and ligustrazine groups(P<0.05)were both higher than that in the model group.The differences were statistically significant.En Face analysis showed that atherosclerotic plaques could be seen in all the full-length aorta of the model group.The percentage of plaques of clopidogrel group and ligustrazine group were lower than that of vehicle group.Notably,ligustrazine had the strongest inhibitory effect on plaque development.Compared with clopidogrel group,the plaque relative area in the ligustrazine group was significantly lower(P<0.05).In terms of platelet activation,compared with the WT group,intracellular cAMP in washed platelets in the model group decreased(P=0.06),and calcium concentration significantly increased(P<0.05),indicating that platelet activation level in this group was increased.The cAMP level in clopidogrel group(P<0.05)and ligustrazine group(P<0.05)was significantly higher than that in model group,while the intracellular calcium concentration in clopidogrel group(P<0.01)and ligustrazine group(P<0.01)was significantly lower than that in model group.Western blot analysis of PI3K/Akt signaling pathway showed no significant difference in PI3K and Akt protein contents among all groups,and the level of phosphorylated Akt(Ser473)was decreased in clopidogrel group,but the difference was not statistically significant.Analysis of phosphorylated Akt levels(Thr308)showed a significant decrease in both clopidogrel and ligustrazine groups(P<0.01).Conclusion:LDLR-/-golden hamsters fed by high-fat and high-cholesterol diet can develop severe hyperlipidemia and atherosclerosis.Ligustrazine,the natural compound,which can activate blood circulation,can inhibit the progression of atherosclerosis and increase the stability of plaques,primarily through its effect on platelet activation via purine receptor signal transduction pathways.