目的 观察CD151及其突变体在大鼠后肢缺血模型中的表达以及对血管新生的影响,进一步研究CD151促血管新生的分子机制.方法 将实验大鼠随机分组,分别给予生理盐水,rAAV-GFP,rAAV-CD151和rAAV-CD151-AAA194-196 (整合素α3/α6结合缺陷突变体),分点注射至大鼠左后肢肌肉组织内,基因转染1周后行股动脉切除术建立大鼠后肢缺血模型,假手术组不切除股动脉.建模5周后测定后肢皮肤温度,检测缺血肌肉组织毛细血管密度,Western blot检测各组大鼠缺血肢体局部CD151的表达.结果 术后5周,rAAV-CD151和 rAAV-CD151-AAA194-196组的CD151蛋白及AAA194-196突变体蛋白表达水平明显高于其余各组(均P＜0.05),rAAV-CD151和rAAV-CD151-AAA194-196两组间蛋白水平无明显差别,但rAAV-CD151-AAA194-196组大鼠缺血后肢平均皮肤温度和毛细血管密度明显低于rAAV-CD151组(均P＜0.05).结论 研究证实CD151促血管新生是通过生成CD151-整合素α3/α6复合体而实现的,CD151-整合素α3/α6复合体的形成是CD151促血管新生的始动机制.%Objective To investigate the effects of CD151 and CD151 mutant gene delivery on angiogenesis in the model of rat hind limb ischemia. Methods Rats were randomly divided into 5 groups- Four groups were injected with saline,rAAV GFP, rAAV CD151 ,or rAAV CD151 AAA194-196 respectively (n = 6 each). After 1 week of transfection,the femoral arteries of the rats were excised to establish the model of left hind limb ischemia. Sham operated animals underwent the same surgical procedure without femoral artery resection. The skin temperature of the hind limb was measured and the capillary density of the ischemic muscle was observed by immunochemical staining for anti CD34 antibody. The expression of CD151 in the ischemic tissue was detected by Western blot. Results The expression of CD151 and CD151 mutant protein was increased significantly in rAAV CD151 and rAAV CD151 AAA194"196 groups as compared with other groups. There was no significant difference in the expres sion between the rAAV CD151 and rAAV CD151 AAA194-196 groups(P>0. 05). The average skin temperature and the capillary density in rAAV CD151 group were higher than those in rAAV CD151 AAA194-196 group. Conclusion The results suggest that CD151 associates directly with α3/α6 integrins to form a functional complexes,thus promotes angiogenesis.
展开▼
