摘要
目的研究survivin基因的反义寡脱氧核苷酸(ASODN)对移植血管内膜增生的影响。方法Wistar大鼠60只,建立自体静脉移植模型,术后随机分为5组:对照组,survivin ASODN 50μg组,200μg组,正义对照组,lipoectin+pluronic组。分别施加不同的处理因素,在移植后1,2周取材。用组织形态学方法比较内膜增生程度;用半定量RT-PCR检测survivin基因的mRNA表达;Western blot检测survivin基因的蛋白产物表达;免疫组化方法检测survivin及增殖细胞核抗原(PCNA)的表达,TUNEL法检测血管平滑肌细胞(VSMC)凋亡。结果静脉移植1~2周内膜增生明显,局部转染50μg survivin ASODN后明显抑制内膜增生(P〈0.05),200μg组受抑制程度较50μg组更为显著(P〈0.05)。静脉移植后,对照组survivin的mRNA及蛋白产物表达显著增加,而survivin ASODN组却显著减少(P〈0.05),VSMC中PCNA表达也同时减少,而TUNEL阳性细胞明显增加。结论survivin ASODNs可显著抑制移植静脉的内膜增生;其作用可能是通过抑制survivin的基因及蛋白产物表达,促进VSMC凋亡而实现的。
Objective To investigate the effect of survivin antisense oligodeoxynucleotides (ASODN) on intimal hyperplasia(IH) in vein graft in rats. Methods Autogenous vein graft model was established in 60 Wistar rats by transplanting the interior jugular vein to common jugular artery using microsurgical technique. The rats were divided into 5 groups according to different processing methods, including survivin ASODN 50μg and 200p4g, scramble ODN 200 μg, control group and lipoectin + pluronic group. Vein graft samples were harvested at 1 or 2 weeks after surgery and histomorphologic methods were used to compare the degree of IH. Survivin mRNA was measured by RT-PCR. Western blotting and immunohistochemistry methods also were used to detect the expression of survivin and PCNA. Apoptosis of VSMC was dectected by TUNEL. Results IH was evident within 1 to 2 weeks after vein grafting, but was markedly inhibited by 50μg of survivin ASODN ( P 〈 0. 05 ) , and even higher inhibition rate in 200μg of survivin ASODN group ( P 〈 0. 05 ). However, no effect was seen in the other group. The expression of survivin mRNA was also inhibited significantly by survivin ASODN. Expression of survivin and PCNA decreased markedly in survivin ASODN group, but the positive cells of TUNEL increased significantly. Conclusions Transfection of survivin ASODN can inhibit the IH of vein graft, which may be effecteol by the inhibition of expression of survivin and enhancement of VSMC apoptosis.
出处
《中国普通外科杂志》
CAS
CSCD
2006年第1期32-36,共5页
China Journal of General Surgery
基金
国家自然科学基金资助项目(30400435
30371401)