| ObjectiveAtherosclerosis(AS)is a degenerative vascular disease based on lipid metabolism disorders and mainly involving large and medium-sized arteries.AS is the main pathological basis of coronary heart disease,cerebral infarction,and other peripheral vascular diseases.Timely prevention and control the development of of AS has very important clinical and practical significance for the health of all mankind.Atherosclerosis(AS)belongs to the categories of "heartache","true heartache",and "juexintong" in traditional Chinese medicine.Qi stagnation,phlegm,and blood stasis are the main pathogenesis.There are often mixed and cause disease,replenishing qi and activating blood,removing blood stasis and removing phlegm,and reducing phlegm and lipid are commonly used in clinical treatment of AS.Traditional Chinese medicine has obvious advantages in the prevention and control of cardiovascular diseases with multiple pathways,multiple targets,and fewer side effects.Danggui Buxue Decoction,as a representative of the TCM method of enriching blood and promoting blood circulation,its therapeutic effect on AS has been confirmed in a large number of clinical and basic studies.This thesis is based on the previous project research,through in vivo animal experiments to study the effect of the effective components of Angelica and Astragalus on AS in ApoE-/-mouse and its specific mechanism,in order to further explore the scientific connotation of Danggui Buxue Decoctions,s anti-AS protective effect,and provides basic experimental data reference for clinical treatment of traditional Chinese medicine.Methods6-week-old male ApoE-/-mice weigh 20-25g were selected and fed with high-fat diet for 8 weeks to established atherosclerosis mice model.The experiment was divided into 18 groups,included the model group(MOD),the total saponins group of astragalus(HZ),the volatile oil group of angelica sinensis(DG),the astragalus total saponins and angelica volatile oil(HG)different ratio group(1:1,2:1,3:1,5:1),the ferulic acid and astragaloside ?(HA)different ratio group(1:1,2:1,3:1,5:1),the ligustilide low and high dose group(LL,LH),the astragaloside ? low and high dose group(AL,AH),the ferulic acid low and high dose group(FL,FH),each group had 10 ApoE-/-mice;At the same time 10 C57BL/6J mice were selected as the normal group(CON),fed with general food.ApoE-/-mice were fed high-fat for 8 weeks,after the AS model was successfully established,from the 9th week,mice in each dosing group were given intragastric administration for 8 weeks.HZ group were given 30 mg/kg of total saponins of astragalus;DG group given 30 mg/kg of angelica volatile oil;The dosage of HG in different ratio groups was calculated based on the extraction rate of total saponins of astragalus and volatile oil of angelica sinensis in different proportions of Danggui Buxue Decoction.The dosage of HG composition(1:1,2:1,3:1,5:1)was 12.33 mg/kg,24.66 mg/kg,36.99 mg/kg,61.65 mg/kg,and the dosage of angelica volatile oil was unified as 24.66 mg/kg;The dosage of HA in different ratio groups was calculated based on the extraction rate of astragaloside ? and ferulic acid in different proportions of Danggui Buxue Decoction.The dosage of HA composition(1:1,2:1,3:1,5:1)was 1.48 mg/kg,2.96 mg/kg,4.44 mg/kg,7.4 mg/kg,and the dosage of ferulic acid was unified as 0.74 mg/kg;Mice in LL and LH group were given 15 mg/kg and 30 mg/kg of ligustilide by gavage;Mice in AL and AH group were given 20 mg/kg and 40 mg/kg of astragaloside? by gavage;Mice in FL and FH group were given 20 mg/kg and 40 mg/kg of ferulic acid by gavage;Mice in the CON and MOD groups were given the same volume of saline intragastrically;The blank group continued to feed with general food,the model group and the drug treat group continued high-fat fed.At the end of 8th/12th week,blood was taken from the tail,and the content of TC,TG,LDL_c and HDL_c in the serum was detected by kit.At the end of 16th week,the eyeballs were removed to take blood,and the heart,aorta,and liver tissues were separated for subsequent detection of various indicators.The content of TC,TG,LDL_c,HDL_c,ET-1,NO,FFA,FC in serum and liver were determined with kit;Oil red O staining was used to detected lipid deposits in liver,aorta,and aortic sinus;HE staining to observed the pathological changes of the liver and aorta;WB detected the expression of lipid metabolism-related proteins in liver;qPCR was used to detected the expression of lipid metabolism genes in liver;Image J software calculated the area of aortic sinus lipid deposition,liver oil red lipid droplets and liver lipid droplets voids;Aortic thickness;The percentage of the aortic sinus and liver lipid deposition area.ResultsAfter 8 weeks of high-fat feeding,compared with the normal group,the serum TC,TG,LDL_c,HDL_c concentrations in the model group were increased(P<0.01).After 12 weeks of high-fat feeding,compared with the normal group,the serum TC,TG,LDL_c,HDL_c concentrations in the model group were increased(P<0.01);Compared with the model group,the content of TC TG and LDL_c in the dosing group were decreased(P<0.05).After 16 weeks of high-fat feeding,compared with the normal group,the content of TC,TG,LDL_c,HDL_c,ET-1,NO,FFA,and FC levels in the model group of serum and liver were increased(P<0.01);Aortic plaque,aortic sinus lipid plasma deposition area were increased(P<0.01);The percentage of aortic sinus lipid deposition area increased(P<0.01);Severe fatty changes in the liver,lipid droplet vacuoles were diffusely distributed,the area of liver oil red lipid droplets and lipid droplet voids increased(P<0.01);The percentage of oil red lipid droplets area in liver was increased(P<0.01);Aortic wall was significantly thickened(P<0.01),accompanied by a large number of inflammatory cell infiltration and cholesterol crystal deposition;The protein expression of ABCA1,SR-BI,APOAI,LXR-?(P<0.05),SREBP2(P<0.01)in liver were decreased;The gene expression of APOAI,ABCA1,ACAT,ABCG5,ABCG8,LCAT,LXR-?,CYP7A1,HMGCR,PCSK9,PPAR-?,SR-BI,SREBP2,LOX-1(P<0.01)in liver were decreased.Compared with the model group,the serum TC,TG,LDL_c(P<0.05),ET-1,NO(P<0.01)levels in the dosing group were decreased;The contents of TC,TG,FFA,FC in liver were decreased(P<0.01);Aortic sinus lipid deposition,liver oil red lipid droplet,liver lipid droplet cavity area were decreased(P<0.05);The percentage of oil red lipid droplets area in liver and lipid deposition areas in aortic sinus were decreased(P<0.05);The protein expression of SR-BI,LXR-?(P<0.05),ABCA1,SREBP2,APOAI(P<0.01)in astragalus total saponins group of liver were increased;The protein expression of ABCA1,SR-BI(P<0.05),LXR-?,SREBP2,APOAI(P<0.01)in angelica volatile oil group of liver were increased;The gene expression of LCAT,CYP7A1,PPAR-?,SR-BI,SREBP2 in the astragalus total saponins and angelica volatile oil group of liver were increased(P<0.01).Conclusion1.The total saponins of astragalus and volatile oil of angelica had a certain intervention and regulation effect on the lipid metabolism of ApoE-/-mice,and the specific onset mechanism might be achieved by regulating the expression levels of lipid metabolism-related proteins in ApoE-/-mice.2.The composition of total saponins of astragalus and volatile essential oil of angelica in different ratios had a certain protective effect on atherosclerosis in ApoE-/-mice,and HG 3:1 was the most effective group.3.The combination of astragaloside ? and ferulic acid in different ratios had a certain protective effect on atherosclerosis in ApoE-/-mice,and HA 5:1 was the most effective group.4.The ligustilide,astragaloside ?,ferulic acid low and high dosage group has a certain protective effect on atherosclerosis in ApoE-/-mice,and the effect of high dose group was better than low dose group. |
PDF Full Text Request