Abstract:Objective:To study the effect of expression of RNA interference targeting protein kinase B (PKB) gene transfection mediated by nanoparticles(NP) on intimal hyperplasia (IH) in vein grafts of rats.
Methods :Nanoparticle PKB short hairpin RNA(shRNA) gene complex was prepared with PLGA and PVA. Autogenous vein graft model was established in 72 rats by transplanting internal jugular vein to carotid artery. The models were randomly divided into 3 groups: (1) PKB shRNA group: PKB-shRNA gene mediated by NP were transfected into the veins before anastomosis. (2) Empty vector group: the veins were transfected by empty vector mediated by NP. (3) Control group (no transfection). The grafted veins were harvested 3, 7, 14 and 28 days after the operation respectively. The mRNA and protein expression of PKB were determined by Northern blot and Western blot. IH was observed by HE and Verhoeff stain. The presence of apoptotic VSMC was detected by TUNEL stain.
Results:Compared to empty vector group and control group, PKB shRNA group had less expression of mRNA and protein of PKB gene(P＜0.05), less IH in the vein graft especially during 7~28d(P＜0.01), and more apoptofic VSMC (P<0.05).
Conclusions:Nanoparticle is an effective gene transfecting carrier, and RNA interference targeting PKB expression can prevent IH and promote VSMC apoptosis after vein grafting.