| Objectives:A rabbit model of in-stent restenosis(ISR)was established by using carotid artery balloon injury combined with high-fat diet.The repair effect of traditional Chinese medicine compound GⅫ on vascular endothelium was observed to evaluate its efficacy.And from the Vascular endothelial growth factor/phosphatidylinositol 3-kinase/protein kinase B(VEGF/PI3K/Akt)signaling pathway to explore the relevant mechanisms.Methods:30 SPF Japanese rabbits were reared for 1 week and randomly divided into 5 groups with 6 rabbits in each group,including sham operation group,operation group,GⅫ group,GⅩⅢ group and atorvastatin group.(1)Sham operation group(normal diet+normal saline):no balloon injury surgery,normal diet,normal saline 4 mL/day by gavage;(2)Operation group(carotid artery balloon injury+high-fat diet+normal saline):carotid artery balloon injury surgery,high-fat diet,normal saline 4 mL/day gavage;(3)GXII group(carotid artery balloon injury+high-fat diet+GⅫ):carotid artery balloon injury surgery,high-fat diet,GXII solution(4g/day)gavage;(4)GⅩⅢ group(carotid artery balloon injury+high-fat diet+GⅩⅢ):carotid artery balloon injury surgery,high-fat diet,GⅩⅢ solution(9g/day)by gavage;(5)Atorvastatin calcium group(carotid balloon injury+high-fat diet+atorvastatin calcium tablets):carotid balloon injury surgery,high-fat diet,atorvastatin calcium solution 5 mg/day by intragastric administration.After 15 days of intervention,rabbits were sacrificed by air embolization,right common carotid artery was extracted,and whole blood was extracted from the aorta.Hematoxylin-eosin(HE)staining was used to observe the morphological and pathological changes of injured blood vessels in each group under microscope.The protein expressions of VEGF,PI3K,Akt,phosphorylated Akt(p-Akt)and endothelial nitric oxide synthase(eNO)in injured carotid vessels were determined by Western blot.Enzyme-linked immunosorbent assay(ELISA)was used to detect the serum levels of endothelin-1(ET-1),total cholesterol(TC),triglyceride(TG),low density lipoprotein(LDL-C)and high density lipoprotein(HDL-C)in rabbits.Results:1.General conditions:rabbits in the sham operation group were in good mental state and could move freely;the other 4 groups were mentally tired and moved relatively slowly after operation in the early stage of modeling.After intervention treatment,the general conditions of rabbits in the GXII,GXIII and atorvastatin calcium groups were improved to varying degrees compared with those before intervention,except that the general conditions of rats in the operation group were not significantly relieved.The mental state of rabbits in the GXII,GXIII and Atorvastatin calcium groups was improved,and the symptoms of bradykinesia were alleviated.2.Histomorphological observation of injured vessels:the intima of the sham operation group was smooth without obvious thickening;compared with the sham operation group,the intima of the vessel in the operation group increased significantly and the lumen area decreased.Compared with the operation group,the thickening degree of the injured intima in the treatment groups was significantly reduced,and the area of the vascular lumen was increased.3.Blood lipid level:compared with sham operation group,serum TC,TG and LDL-C levels in operation group were increased(P<0.05),while HDL-C level was decreased(P<0.05);Compared with the operation group,the levels of serum TC,TG and LDL-C in the treatment groups were decreased(P<0.05),while the level of HDL-C was increased(P<0.05).4.Protein expression in injured vascular tissue:Compared with the sham operation group,the protein expressions of VEGF,P13K,p-Akt and eNOS in the operation group were up-regulated(P<0.01)and down-regulated(P<0.01).Compared with the operation group,the expression of tissue proteins VEGF,P13K and p-Akt in each treatment group was stable(P<0.01),and eNOS expression was up-regulated(P<0.01).5.Serum ET-1 level:Compared with sham operation group,serum ET-1 level of operation group was increased(P<0.01);Compared with the operation group,the serum ET-1 level in the treatment groups was decreased(P<0.01).Conclusions:GⅫ prescription(GⅫ,GⅩⅢ)can reduce the thickening degree of intima,regulate the levels of TC,TG,LDL-C and HDL-C of blood lipids in the rabbit model of carotid artery balloon injury,and has the protective effect of vascular endothelium.Its effect may be related to VEGF/PI3K/Akt signaling pathway.GⅫprescription can improve vascular endothelial function and reduce intimal hyperplasia by regulating VEGF,PI3K and p-Akt expression,upregulating eNOS expression and decreasing ET-1 level. |
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