Effects Of Astragalus And Angelica On The Extracellular Matrix And Endothelial Cell Injury In Rabbit Model Of Intimal Hyperplasi

Objective:The rabbit common carotid artery vascular intimal hyperplasia model was established and Astragalus and Angelica were used to intervene in different ratios to observe their effects on vascular intimal hyperplasia,and the effective compatibility ratio of Astragalus and Angelica against vascular intimal hyperplasia was established.Based on the effective compatibility of anti-vascular intimal hyperplasia,extracellular matrix（ECM）and vascular endothelial cells（VEC）are used as targets to explore the effects of the compatibility of Astragalus and Angelica on ECM deposition and VEC damage in the vascular intimal hyperplasia model.Methods:1,Thirty New Zealand white rabbits,half male and half female,were randomly divided into sham operation group,high fat+unilateral cannula group（divided into 2 weeks and 3 weeks subgroup according to modeling time）and hyperlipidemia+bilateral cannula group（divided into 2weeks and 3 weeks subgroups according to modeling time）.In the unilateral cannula group,a silicone rubber tube（20 mm in length and inner diameter1.0 mm）was placed on the lateral side of the left common carotid artery and fixed,while in the bilateral cannula group,a silicone rubber tube was placed on the lateral side of the bilateral common carotid artery and fixed.Both groups were fed with 2%cholesterol diet for 2-3 weeks the day after operation.In the sham operation group,the common carotid artery was separated by operation,no vascular cannula was performed,and the rabbits in the sham operation group were fed with common feed.At the 2nd and 3rd week after operation,the pathomorphological changes of the injured common carotid artery were observed,the index of intimal hyperplasia was measured by morphometry,and the expression ofα-smooth muscle actin（α-SMA）,osteopontin（OPN）,type I collagen（Col-Ⅰ）,inflammatory response factor interleukin-1β（IL-1β）,tumor necrosis factor-α（TNF-α）in intimal hyperplasia was determined by immunohistochemical method.2,Ninety six New Zealand white rabbits are half male and half male.Based on the above determination of the conditions for establishing vascular intimal hyperplasia,they were randomly divided into sham operation group（group A）,12 rabbits,model group（group B）,12 rabbits,Astragalus alone group（group C,1g/kg）,Angelica alone group（group D,1g/kg）12rabbits,Astragalus-Angelica of 1:1 combination group（group E,Astragalus1g/kg+Angelica 1g/kg）12 rabbits,Astragalus-Angelica of 1:5 combination group（group F,Astragalus 1g）/kg+Angelica 5g/kg)12rabbits,Astragalus-Angelica of 5:1 combination group（group G,Astragalus5g/kg+Angelica 1g/kg）12rabbits and atorvastatin group（group H,5mg/kg）.A rabbit model of bilateral intimal hyperplasia of common carotid arteries was established,and high-fat diet was continuously fed from the first day after modeling.The sham operation group was continuously fed regular feed from the first day after surgery.The rats were given intragastric administration on the first day after modeling,once a day,after 28 days of continuous administration,blood was collected from the abdominal aorta and the animals were sacrificed to take the injured common carotid artery.Immunohistochemical method was used to detect the expression of Collagen 1（Col-I）,Fibronectin（FN）,Matrix metallopeptidase 2（MMP-2）and Tissue inhibitor of metalloproteinases 1（TIMP-1）in the intima of the common carotid artery in rabbits.ELISA method was used to determine plasma endothelin-1（ET-1）and Prostacyclin（PGI₂）levels.Nitrate reductase colorimetry was used to detect plasma Nitric oxide（NO）levels.Western blot was used to determine the expression of Endothelial nitric oxide synthase（e NOS）protein in the injured common carotid artery.Results:1,After intervention with high fat and silicone rubber tube for 2and 3 weeks,intimal hyperplasia was found in common carotid artery.Intimal area（IA）,intimal thickness（IT）,intimal area proliferation rate（HRIA）and intimal thickness（HRIT）increased significantly.Intimal hyperplasia in bilateral cannula was more obvious than that in unilateral side,and intimal hyperplasia in 3 weeks was more serious than that in 2weeks.After 3 weeks of bilateral cannula,the expression ofα-SMA in vascular intima was significantly reduced,and the expression of OPN,Col-Ⅰ,IL-1β,and TNF-αwere significantly increased.2,Compared with the model group,Astragalus alone group,Angelica alone group,Astragalus-Angelica of 1:1 combination group,Astragalus-Angelica of 1:5 combination group,Astragalus-Angelica of 5:1 combination group were inhibited the expression of Col-I,FN and MMP-2 and enhanced the expression of TIMP-1 in hyperplastic intima.In the Astragalus-Angelica compatibility,the Astragalus-Angelica 5:1 compatibility is equivalent to the1:1 compatibility.Compared with the model group,Astragalus alone group,Angelica alone group,Astragalus-Angelica of 1:1 combination group,Astragalus-Angelica of 1:5 combination group,Astragalus-Angelica of 5:1 combination group were Significantly increased the levels of NO in plasma（P<0.05）,the expression of e NOS protein in the injured blood vessel was significantly reduced（P<0.05）,and the content of plasma ET-1 was significantly reduced（P<0.05）.Compared with the model group,Angelica alone group,Astragalus-Angelica of 1:1 combination group,Astragalus-Angelica of 1:5combination group,Astragalus-Angelica of 5:1 combination group were Significantly increased the levels of PGI2 in plasma（P<0.05）.In the Astragalus-Angelica compatibility,the Astragalus-Angelica 5:1 com patibility is equivalent to the 1:1 compatibility.Conclusion:The model of intimal hyperplasia of the common carotid artery in rabbits was successfully replicated.It is advisable to intervene with bilateral common carotid artery cannula for more than 3 weeks.The compatibility of Astragalus and Angelica has a significant inhibitory effect on intimal hyperplasia after VEC is damaged.Its effect may be related to the inhibition of extracellular matrix ECM deposition in the proliferative intima and the improvement of VEC dysfunction.