| Research purposes:Percutaneous coronary intervention is facing the challenge of instent restenosis.Traditional Chinese medicine believes that the combination of toxin and blood stasis is one of the main pathogenesis of in-stent restenosis,and the method of detoxicating and activating blood circulation is the main therapeutic principle to inhibit instent restenosis.Endothelial injury is the trigger of in-stent restenosis and the pathological basis of the formation of toxin and blood stasis.Therefore,the purpose of this study is to study the effect of Detoxification and Blood Circulation Decoction on the endothelialization of injured vessels and the activities of related cytokines NO and ET-1 in the process of re-endothelialization,and to understand how Detoxification and Blood Circulation Decoction affects the expression of VEGF protein to promote the re-endothelialization of injured vessels and prevent and treat in-stent restenosis,so as to provide a new idea for the prevention and treatment of in-stent restenosis with traditional Chinese medicine.Methods:72 SD rats were randomly divided into 6 groups: shame-operation group,model group,small,medium and large dose group of Detoxification and Blood Circulation Decoction and western medicine control group(atorvastatin calcium tablet group).The rats in the shame-operation group were only ligated,and the rats in other groups were injured by balloon.The model group and the sham-operation group were gavaged with normal saline,and the other groups were gavaged with corresponding doses for 14 days and 28 days respectively.HE staining was used to detect the pathological changes of thoracic aorta of rats in each group;Evans blue staining was used to evaluate the efficiency of re-endothelialization;The contents of vasoactive factors NO and ET-1 were measured in each group;Western blot was used to detect the expression of VEGF protein in the vascular tissue of thoracic aorta.Results:1.HE staining results showed that the overall structure of the aorta in the shame-operation group was basically normal,the vascular endothelial structure of the tissue was smooth,and the connection between endothelial cells was tight.In the model group,the intima of blood vessels was thickened,as shown by the yellow arrow in the figure.The smooth muscle cells in the vascular wall of the tissue proliferated and arranged in disorder.The tissue showed obvious infiltration of inflammatory cells,and the lumen area was narrow.The overall structure of aorta in Detoxification and Blood Circulation Decoction group and Atorvastatin calcium tablet group was slightly abnormal,the vascular endothelial structure was rough,the endothelial cells were tightly connected,the vascular intima was slightly thickened,but there was no obvious inflammatory cell infiltration.2.Evans blue staining was used to evaluate the results of reendothelialization: Evans blue staining was used to evaluate the effect of Detoxification and Blood Circulation Decoction on promoting the reendothelialization of damaged blood vessels.The part stained blue indicated cell damage,without endothelial cell coverage.Compared with the sham-operated group,the endothelialization rate in the model group decreased significantly(P<0.001)Compared with the model group,the re-endothelialization rate of each group of Detoxification and Blood Circulation Decoction was significantly higher,among which the middle and high dose groups of Detoxification and Blood Circulation Decoction and the atorvastatin calcium tablet group were statistically significant compared with the model group(P<0.05).3.Determination of NO content in serum: compared with the sham group,the expression of NO in serum in the model group was significantly reduced(P<0.001).Compared with the model group,14 days after modeling,the serum NO level in the middle and high dose groups and atorvastatin group increased significantly(P<0.01),while there was no significant difference in the low-dose group,and 28 days after modeling,the serum NO level in the Detoxification and Blood Circulation Decoction group and atorvastatin group increased significantly(P<0.05).There was no significant difference between the high-dose group of Detoxification and Blood Circulation Decoction and the atorvastatin calcium tablet group(P>0.05).4.Determination of ET-1 content in serum: compared with the sham group,the expression of ET-1 in serum in the model group was significantly increased(P<0.001).Compared with the model group,the serum level of ET-1 decreased significantly in the 14-day Detoxification and Blood Circulation Decoction high-dose group and atorvastatin group(P<0.01),but there was no significant difference in the decrease between the medium-and low-dose groups.The serum level of ET-1 decreased significantly in the 28-day Detoxification and Blood Circulation Decoction in the middle and high dose groups and atorvastatin group(P<0.05),but there was no significant difference in the low dose group.There was no significant difference between the high-dose group of Detoxification and Blood Circulation Decoction revitalization the atorvastatin calcium tablet group(P>0.05)5.Measure the expression of VEGF in vascular tissue.The results showed that the expression of thoracic aorta VEGF in the model group was significantly reduced compared with the sham surgery group(P<0.001).Compared with the model group,the expression of VEGF in each group of Detoxification and Blood Circulation Decoction increased to different degrees,among which the medium and high dose group and atorvastatin calcium tablet group were statistically significant compared with the model group(P<0.05),and the high-dose group of Detoxification and Blood Circulation Decoction had a significant effect with atorvastatin calcium tablet group(P<0.001),in addition,the difference between the high-dose group of Detoxification and Blood Circulation Decoction and atorvastatin calcium tablet group was not significant and there was no significant significance.Conclusion:1.Detoxification and Blood Circulation Decoction can improve the pathological changes of damaged blood vessels,promote endothelialization of damaged blood vessels;2.Detoxification and Blood Circulation Decoction can regulate vascular endothelial factors and restore vascular endothelial function;3.Detoxification and Blood Circulation Decoction can promote the expression of VEGF protein,promote endomembrane repair,accelerate endothelialization,and reduce vascular stenosis in the stent. |
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