Study On Genes Related To Restenosis Of Rabbit Vein Grafts Based On Transcriptome Sequencing

Background:Coronary atherosclerotic heart disease is the leading cause of human mortality[1,2],and the incidence of coronary heart disease is increasing in our patients[3].Coronary artery bypass grafting(CABG)is crucial in the surgical treatment of coronary artery disease with relatively good long-term results.The internal mammary artery(Internal Mammary Artery IMA)or the saphenous vein(Saphenous Vein SV)is often chosen as the bypass vessel for the surgery.The arterial bridge vessel has the best long-term patency rate,but its length is limited and cannot meet the clinical demand.Unlike arterial bridges,the long-term patency rate after venous bridging is relatively low,with an incidence of occlusion of approximately 10%—30%within 1 year[4-6];an annual increment of approximately 2%in years 1-6 and 4%in years 6-10;after 10 years,approximately 40%of SV bridges will be completely occluded,with varying degrees of stenosis in the remaining 50%of vessels that are not completely occluded[7,8].However,multiple factors and mechanisms contribute to venous bridge vasculopathy,and different factors may be present at different stages[9].The main influencing factors are surgical injury,hemodynamic changes,oxygen partial pressure alterations,and the properties of the venous vessels themselves,but the mechanisms associated with restenosis of venous bridge vessels at the genetic level are still less studied.Objective:By constructing a New Zealand rabbit external jugular vein-common carotid artery vascular graft model,Confirm the feasibility of animal model.we simulated the use of saphenous vein coronary artery bypass surgery in patients with coronary atherosclerotic heart disease and examined the intima and intima of the bridge vein vessels after grafting,and then explored the genes that might be associated with the process of bridge vessel restenosis in the grafted veins by transcriptome sequencing technology(RNA-seq).Methods:In this experiment,24 experimental New Zealand rabbits,weighing 2-2.5 kg,were selected.ipsilateral external jugular vein graft to common carotid artery animal model was established using continuous suture method.The intimal thickening of the venous bridge was assessed by hematoxylin-eosin staining in each group.RNA high-throughput sequencing(RNA-seq)was used to identify genes differentially expressed in the grafted venous vessels and normal venous tissues,and genes with differential expression were analyzed and collated using GO and KEGG databases.Results:1.In the experimental group,compared with the control group,the intima and mesentery of the bridge vessels were thickened at 14 days postoperatively,and at 28 days postoperatively,the intima and mesentery of the bridge vessels were thickened more significantly.2.RNA high-throughput sequencing technology was used to detect specimens at 28 days postoperatively versus the control group and analyze the differences between the groups.2196 differential genes were finally obtained between the two groups,including 1583 up-regulated genes and 608 down-regulated genes.GO as well as KEGG analysis was used to predict the biological functions of differentially expressed genes,some of which and pathways have been confirmed to be related to the process of bridging vein restenosis.These include cytosolic protein backbone binding,GTPase binding agonism,calcium binding,cytokine receptor activity,and leukocyte activation-adhesion.Based on the results of differential analysis,a differential intergenic network analysis was performed to screen the top 20 genes in central position CD4,SYK,CD19,PIK3CD,ZAP70,CD28,CCL5,IL7R,CXCR4,PTPRC,CCR1,CD2,IL2RG,CLEC7A,CLEC4E,HLA-DMA,ITK,NTRK1,CCR5,PTPN6.Ten hub genes were further screened,which were CD4,ZAP70,SYK,CD28,PIK3CD,CXCR4,CCR5,ITK,CCL5 and BTK.Each gene was retrieved and screened through GeneCards database,and then the biological function of each gene was screened and evaluated.You end up with SYK and ZAP70 Hub genes.Conclusion:Animal models were successfully established,and vascular pathology sections at 14 and 28 days postoperatively suggested significant graft vessel thickening.Transcriptome sequencing of experimental vein bridge specimens and control specimens at 28 days postoperatively was performed,and analysis revealed some differential genes that may be involved in the development of vein graft restenosis.However,further studies are needed to investigate the mechanism of the effect of related genes on intimal proliferation and to provide more methods for the treatment of vein graft restenosis.