重组arresten蛋白对自体移植静脉内膜增生的抑制作用

目的 观察原核表达人arresten重组蛋白对自体移植静脉内膜增生的抑制作用.方法 用pRSET原核表达系统表达并纯化人arresten重组蛋白.将Wistar大鼠颈外静脉移植于腹主动脉,建立大鼠自体静脉移植模型,实验分3组:假手术组、移植对照组和移植实验组.自术后第3天起,皮下注射给予arresten重组蛋白(每日4 mg/kg体重)处理.4周后取移植静脉组织标本,进行病理组织学观察与免疫组织化学染色分析.结果 移植组移植静脉均呈现典型的内膜增生、肥厚,导致血管管腔狭窄;新生内膜主要有过度增殖的α-SMA染色阳性平滑肌细胞组成.移植实验组移植静脉内膜增生受到明显抑制,新生内膜面积(0.12±0.07)mm2及新生内膜/中膜面积比(0.373±0.085)均显著低于对照组(0.38±0.11)mm2,1.621±0.086,P<0.01];并且实验组移植静脉新生内膜细胞PCNA标记指数显著低于对照组(15.62±3.97)%比(56.36±3.49)%,P<0.01].结论 重组arresten蛋白通过抑制新生内膜平滑肌细胞的增殖能有效抑制自体移植静脉内膜增生的发生发展,在防治血管重建术后再狭窄方面显示出良好的应用前景.

Inhibitory effect of arresten recombinant protein on neointimal hyperplasia of venous autografts In a rat autogenous vein graft model

Objective To investigate the inhibitory effect of prokaryotie-expressed arresten recombinant protein on neointimal hyperplasia of autogenous vein graft in rats. Methods Human arresten recombinant protein was expressed and purified from pRSET prokaryotic expression system. The model of autogenous vein graft was prepared by transplanting the external jugular vein into aorta in Wistar rats by means of micro-surgery. The rats were divided into three groups : sham operation group, autograft control group,autograft experimental group. From 3rd day after transplantation,arresten protein was injected intra-peritoneally daily at a dosage of 4 mg/(kg·d).Four weeks after treatment,venous autografts were removed at autopsy and subjected to histopathological examination and immanohistochemical staining. Results In autograft control and experimental groups,the venous autografts displayed well-developed neointimal proliferative lesions composed of a large number of smooth muscle cells that stained positive for α-SMA. The neointimal hyperplasia resulted in vascular lumen stricture in venous autografts. In autograft experimental group,the neointimal hyperplasia was suppressed obviously, and its neointimal area (0.12 ±0.07) mm2 and NIA/MA ratio (0.373 +0.085 ) of venous autografts were significantly reduced as compared with those in control group (0.38 + 0.11)mm2, 1.621±0.086 (P<0.01). And its PCNA labeling index of neointimal cells was significantly lower than that in control group (15.62±3.97) % vs (56.36±3.49) % (P<0.01). Conclusion Recombinant arresten protein could significantly inhibit the proliferation of neointimal smooth muscle cells and the development of neointimal hyperplasia of venous autografts in rats. Arresten is a powerful inhibitor of restenosis after vascular reconstructive operation with a potential for therapeutic use.