A Whole Transcriptome Study On Gene Expression Associated With Intimal Hyperplasia Of Vein Graft In Rats

BackgroundAt present,China's population is aging seriously.The incidence of arteriosclerosis occlusive diseases of lower limbs is increasing year by year,which constantly threatens the life and health of human beings.Peripheral arterial stenosis or sclerosis occlusion is the main cause of severe lower limb arterial ischemia and distal limb necrosis.Surgical treatment can restore the blood supply to an injured limb to cure an ischemic limb.Autologous vein transplantation is one of the important surgical treatment methods to open the arterial blood supply of the affected limb,which can greatly improve the prognosis of patients.But the long-term effect is not ideal because of the restenosis of the vascular lumen.Intimal hyperplasia is the main cause of restenosis after vein transplantation.lnc RNA,circ RNA,micro RNA and m RNA all play important roles in intimal hyperplasia,but the expression of RNA at the whole transcriptome level in the process of vein intimal formation has not been reported in detail.ObjectiveThe rat model of autologous vein transplantation was constructed to detect theintimal hyperplasia and cell proliferation of the transplanted vein,and the regulatory mechanism in the process of intimal hyperplasia of the transplanted vein was discussed by means of RNA sequencing.MethodsIn this study,the left external jugular vein of rats was transplanted to the abdominal aorta by intermittent suture to construct a rat model of autologous vein transplantation.Hematoxylin-eosin staining,immunohistochemical staining and transmission electron microscopy were used to evaluate the intimal hyperplasia of the transplanted vein.We further used RNA high-throughput sequencing to identify differentially expressed RNA in transplanted vein tissues,mainly including m RNA,long non-coding RNA(lnc RNA),micro RNA(m RNA)and circ RNA(circ RNA).We used GO and KEGG analysis to predict the function of differentially expressed RNA.Pearson correlation coefficient and Mi Randa database were used to predict competitive endogenous RNA(ce RNA)pairs.ResultsThe experimental results of the rat autologous vein transplantation model showed that the graft vein intima hyperplasia was obvious in the 14 days after the operation compared with the control group.The differential expression of 4,223 m RNA,2221 lnc RNA,117 circ RNA and 265 micro RNA were found by RNA high-throughput sequencing in the two groups,and the ce RNA relationship pairs of lnc RNA-micro RNA-m RNA and circ RNA-micro RNA-m RNA were predicted.GO and KEGG pathways predicted the potential function of differentially expressed RNA.They were widely involved in inflammatory response,cell proliferation,and cell phenotypic regulation pathways,including c GMP-PKG,calcium regulation and cell adhesion.ConclusionAbove results show that the vein intima of the graft had obvious hyperplasia after14 days compared with control group.High-throughput sequencing technology analysis and bioinformatics analysis were used to identify RNA differentially expressed between the control group and 14 days after surgery group.some of the RNA including m RNA,mi RNA,lnc RNA,circ RNA may be involved in cell proliferation,inflammatory response related signaling pathways.These differentially expressed RNA may through these signaling pathways play a key role in the progress of the transplant neointimal hyperplasia.However,further functional experiments are still needed to explore the specific pathogenesis of the involvement of these differentially expressed RNA in the transplantation of intima hyperplasia.It is expected to provide basic theoretical support for the clinical diagnosis and treatment of arteriosclerosis obliterans of the lower extremity.