| Purposes: To observe the changes of aortic protein chip in Apo E-/-AS mice,and the intervention of Er Chen decoction and Taohong Siwu decoction.Investigate the effect and mechanism of Er Decoction and Taohong Si Wu Decoction on atherosclerosis.Material and methods: 20 C57BL/6J mice were served as blank control group,and normal mice were fed with feed,60 Apo E-/-mice served as model group AS and were fed a high-fat diet for model replication,8 weeks later,60 model mice were randomly divided into model group 20,low dose group 10,medium dose group 10,high dose group 10 and positive control group 10,The model group and the treatment group continued to be fed high fat diet for 4 weeks,Low,medium and high dose groups were given containing crude drug for traditional Chinese medicine decoction 0.72g/m L,1.44g/m L,2.89g/ m L gavage.The positive control group was given intragastric administration of Xuezhikang Capsule with concentration of 0.0078g/m L.Intragastric administration of 0.2m L in the morning and evening,normal group and model group were treated with distilled water of equal volume.12 weeks later,The formation of aortic plaques in normal group and model group was observed under ultrasound,The blood lipid levels of mice in each group were detected by automatic biochemical analyzer,HE staining and oil red O staining were used to observe the pathological changes and lipid deposition of the aorta in each group of mice,The levels of serum TNF-?,IL-6,NO and ET-1 in each group were detected by Elisa,The expression of protein in aortic tissue of model group mice was analyzed by protein chip technique,to screen the possible pathogenesis of AS.The differentially expressed proteins were screened from the protein level or gene level by Western Blot and RT-PCR techniques.Results: 1.Ultrasound results showed,compared with the blank control group,there was a large area of plaque formation in the intima of the model group mice.2.Blood lipid levels showed,compared with the blank control group,model group,serum TC,TG and LDL levels were significantly higher(P<0.01),the level of HDL was significantly decreased(P<0.01);compared with the model group,Chinese medicine low,medium and high dose group and positive control group,TC,TG,LDL were significantly decreased(P<0.05 or P<0.01),the level of HDL was significantly(P<0.01).3.HE staining showed that compared with the blank control group,model group mice have larger aortic plaque formation,compared with the model group,the treatment group were changed in different extent,with the high dose of traditional Chinese medicine and the positive control group is better.4.Oil red O staining showed that compared with the blank control group,model group,mouse aortic lumen with large lipid deposits.5.Elisa test results showed that compared with the blank control group,model group mice serum TNF-?,IL-6 and ET-1 levels were significantly higher(P<0.01),the level of NO was significantly decreased(P<0.01);compared with the model group,Chinese medicine low,medium and high dose group and positive control group TNF-?,IL-6 alpha and ET-1 were decreased significantly(P<0.05 or P<0.01),NO level increased significantly(P<0.05 or P<0.01),NO/ET-1 increased significantly(P<0.05 or P<0.01);compared with the positive control group,low dose group of Chinese medicine,TNF-?,IL-6,ET-1,alpha NO/ET-1 level increased significantly(P<0.05 or P<0.01),the level of NO was significantly decreased(P<0.01),no significant difference between the high dose group.6.Protein microarray results showed that compared with the blank control group,protein up regulated,including 31 proteins up-regulated more than 15%,27 proteins were up-regulated more than 20%;72 protein down regulated,of which 27 protein decreased more than 15%,18 proteins down more than 20%.Screening showed that the expression of TGF-?/PI3K/AKT/e NOS signaling pathway related protein significantly decreased expression of TNF-?/HIF-1?/NF-?B signaling pathway related protein increased significantly.7.The Western blot results show that compared with the blank control group,the expression of TGF-?,PI3 K protein and phosphorylation of e NOS,AKT protein was significantlydecreased(P<0.01),the expression of Nox4 and NF-?B protein was significantly increased(P<0.01),in model group;compared with the model group,the expression of TGF-?,PI3 K,Nox4,NF-?B and AKT,e NOS protein phosphorylation level was improved in different degrees,with Chinese medicine dose group,high dose of Chinese medicine group,positive control group(P<0.05 or P<0.01)effect is more obvious.8.RT-PCR results showed that compared with the control group,the expression levels of model mice aorta TGF-? and PI3 K m RNA decreased significantly(P<0.01),VEGF,MMP-9,HIF-1? m RNA expression level was significantly increased(P<0.01);compared with the model group,the treatment group of mice aorta TGF-?,PI3 K,VEGF,MMP-9,HIF-1? m RNA the expression level was improved in different degrees,with Chinese medicine dose group,high dose of Chinese medicine group,positive control group(P<0.05 or P<0.01)effect is more obvious.Conclusion: 1.The expression profiles of aortic protein in Apo E-/-atherosclerotic mice changed markedly after 12 weeks of feeding with high fat diet.2.Erchen decoction and Taohongsiwu Decoction can effectively improve blood lipid level,inflammatory reaction and protect aorta injury in Apo E-/-atherosclerotic mice.3.The mechanism of Erchen decoction and Taohong Siwu Decoction protects Apo E-/-atherosclerotic mice from aortic injury may be related to the activation of TGF-?/PI3K/AKT/e NOS signaling pathway related proteins.4.The mechanism of Erchen decoction and Taohong Siwu Decoction protects Apo E-/-atherosclerotic mice from aortic injury may be related to the inhibition of TNF-?/Nox4/NF-?B/HIF-1?signaling pathway related protein expression. |
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