| Summary OBJECTIVE: To establish an animal model of white rabbit AS and observe the mechanism of Yiqi Huoxue Recipe on anti-inflammatory and stable plaques in AS rats,and to detect related indicators,namely,lipid,NF-?B,VCAM-1,MCP-1,and endothelin.And the impact of the fibrinolytic system.Through experiments,the effect and mechanism of Yiqi Huoxue Recipe on the anti-inflammatory and stable spots produced by AS were explored,and the theoretical experimental basis for the clinical application of traditional Chinese medicine for prevention and treatment of AS was provided.Materials and Methods: Modeling and grouping of experimental animals: Animals were randomly divided into two groups after one week of adaptive feeding: 8 in the normal control group and fed standard rabbit feed;28 in the surgical group fed with high fat diet.Four weeks later,white rabbits from 28 surgical groups were first subjected to carotid artery balloon injury,and then fed with high-fat diet.New Zealand white rabbits survived after surgery were randomly divided into model group(8 rats)and Qiqi.There were three groups: Huoxue Compound Group(8)and simvastatin group(8).In the experimental administration group,only the drug feed was fed every morning,waiting for the feed to be completed,and sufficient high-fat diet(120 g per day)was fed;the model group was fed with high-fat diet;the normal group was only given Ordinary feed feeding.In the experimental group,the white rabbits were weighed once a week,and the doses of the white rabbits in each group were adjusted according to the weight of the white rabbits.The total administration time was 4 weeks.1.Thirty-six male New Zealand white rabbits with a healthy weight of 2.5±0.5 kg were fed for 1 week,and 4 of them were randomly selected as normal groups,and the experimental feeding was performed with ordinary basic rabbit feed.The remaining rabbits underwent balloon injury of the common carotid artery.After the operation,the rats were fed with high-fat diet for four weeks.The total number of deaths of the rabbits during the operation and after surgery was four.The other 24 white rabbits were randomly divided into a model group(n=8),a simvastatin group(n=8),and a Yiqihuoxue compound group(n=8)for 4 weeks.The experimental blood collection was carried out at the 0th,4th,and 8th weekends of the experiment,and after the experimental animals were sacrificed at the 8th weekend,the materials were taken and sliced.2.To observe the histopathological changes of the damaged blood vessels in the 8th weekend of each group by hematoxylin-eosin(HE)staining and the establishment of the rabbit model of the total neck of the rabbits with balloon injury,and the method of supplementing qi and activating blood to the damaged blood vessels.Evaluation of membrane proliferation and cell structure;3.The levels of TG,TC,HDL-C and LDL-C in plasma of each group of animals at the 0th,4th and 8th weekends were detected by automatic biochemical analyzer.4.RT-PCR was used to detect the expression of NF-?B,VCAM-1 and MCP-1 mRNA in carotid arteries.5.Immunohistochemistry was used to determine the expression of NF-?B,VCAM-1 and MCP-1 proteins in the carotid arteries of each group.6.Spectrophotometric determination of plasma nitric oxide(NO)levels at week 8;7.Determination of plasma endothelin-1(ET-1)content at the 8th week using the ELISA method;8.The plasma levels of t-PA,PAI-1 and TF at the 0th week,4th week and 8th week of each group of rabbits were detected by ELISA.Results: 1 confirmed the successful replication of the AS model,and Yiqihuoxue compound has a significant inhibitory effect on intimal hyperplasia.2 At the time of enrollment,there was no difference in blood lipid levels and body weight of each group of animals and they were in a normal distribution(expressed as 0 weeks).At the end of the fourth week,the levels of TC,TG,and LDL-C in the plasma of rabbits fed with high fat were calculated.The ratios of TC/HDL-C and LDL-C/HDL-C were higher than those of the blank control group,and the difference was significant(P <0.01),the amplitude of LDL-C and TC changes were more obvious in 4 cases;HDL-C was lower than the blank control group(P<0.05);Yiqihuoxue compound had a good lipid-lowering effect,in the experiment At the end of the 8th week,TC,TG,and LDL-C were reduced by 38.3%,31.6%,and 50.1%,respectively,and HDL-C was increased by 160.9%,and TC/HDL-C and LDL-C/HDL were decreased.The-C ratio was 62% and 89%.Simvastatin was significantly lower in TC and LDL-C than in Yiqi Huoxue Recipe(P<0.05).Yiqi Huoxue Recipe has obvious advantages in raising HDL-C(P>0.05).There was no significant difference in TG between the two groups(P>0.05).3 Yiqi Huoxue Recipe has significant inhibitory effect on the level of inflammatory factors: at the 8th week of the experiment,the levels of NF-?B,VCAM-1 and MCP-1 mRNA in the carotid artery of each group decreased in the control group.The content of the model group was the highest,which was statistically significant compared with the simvastatin group and the Yiqihuoxue compound group(P<0.05).NF-?B and VCAM-1 were between the Yiqihuoxue compound group and the simvastatin group.The difference of MCP-1 mRNA level was not statistically significant.Compared with the model group,the NF-?B,VCAM-1,and MCP-1 proteins in the Yiqihuoxue compound experimental group were significantly decreased,and the difference was statistically significant(P<0.O5),and there was no significant difference in the expression of NF-?B,VCAM-1 and MCP-1 between the Yiqi Huoxue Recipe group and the simvastatin group(P>0.05).4.Yiqi Huoxue Recipe can reduce the secretion of ET-1 while promoting the secretion and synthesis of NO.It can promote the balance of NO/ET-1 in vivo and protect EC: at the 8th weekend of the experiment.The plasma NO level in the model group was significantly lower than that in the normal group,and the difference was significant(P <0.01).The plasma NO level in the simvastatin group and the Yiqihuoxue compound group increased compared with the model group.Significantly(P<0.01);the level of plasma NO in rabbits in the Yiqihuoxue compound group was significantly higher than that in the simvastatin group(P<0.01);at the 4th week after balloon injury,ELISA was used.The ET-1 content in the plasma of the white rabbits in the model group was significantly increased compared with the normal group.By comparing with the model group,it was possible to obtain the plasma ET-1 of the white rabbits in the Yiqihuoxue compound group and the simvastatin group.The level of decrease was significant(P<0.05),but there was no significant difference between the two groups(P>0.05).5 balloon damage can not only affect the stability of the fibrinolytic system in the body,and can be conducive to the expression of tissue factors,causing thrombosis in the body,Yiqihuoxue compound can stabilize the fibrinolytic system,and play an anticoagulant,antithrombotic The effect: In the 8th week of the experiment,the PAI-1 content in the model group continued to rise.On the contrary,the plasma PAI-1 content of the experimental group using Yiqihuoxue compound and simvastatin could be reduced by 48% and 44.7%,respectively,compared with the model group.There was significant difference(P<0.01),but there was no significant difference in the PAI-1 content between simvastatin and Yiqihuoxue compound group.At the 8th week of the experiment,there was no significant difference in the t-PA content in the normal control group compared with the 4th week.The t-PA content in the model group was significantly lower than that at the 4th week(P<0.05),and it was able to significantly Compared with the normal control group,the content of t-PA in the simvastatin group and the Yiqihuoxue compound group was significantly higher than that in the model group,and the difference was significant(P<0.05).Compared with the two groups,the plasma t-PA content was significantly higher than the latter(P<0.05).The TF content in each period was measured.At the 8th weekend of the experiment,the TF content in the simvastatin group and the Yiqihuoxue compound group was significantly lower than that in the model group(P<0.01).There was no significant difference between the two groups(P>0.05).).The level of TF antigen in tissue homogenate was detected by ELISA.The results showed that at the 8th week,the carotid TF level in the model group was significantly higher than that in the normal group(P<0.01).conclusion: 1.Yiqihuoxue compound has a significant inhibitory effect on intimal hyperplasia.2.Yiqi Huoxue Recipe can cause the decrease of plasma TG,TC and LDL-C content in rabbits with experimental hyperlipidemia,and can improve the level of HDL-C in plasma,which has a better promoting effect on lipid regulation.3.Yiqi Huoxue Recipe can inhibit the activation of NF-?B,VCAM-1 and MCP-1,reduce the level of inflammatory factors,stabilize plaque and prevent the formation of arteriosclerosis.4 Yiqi Huoxue Recipe can promote the secretion and synthesis of NO in plasma and inhibit the expression of ET-1,which is beneficial to restore NO/ET-1 balance and has a good protective effect on endothelial cells.5 Yiqi Huoxue Recipe can balance the fibrinolytic system in the body,inhibit the release of tissue factor,and have anticoagulant and antithrombotic effects. |
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