钙调神经磷酸酶信号通路及负调控信号糖原合成酶-3β对大鼠经皮血管成形术后再狭窄的作用

Effects of CaN-NFATc and signal GSK-3β on the restenosis following balloon injury in rats

目的研究钙调神经磷酸酶(CaN)-活化T细胞核因子(NFAT)信号通路及此通路抑制因子糖原合成酶-3β(GSK-3β)在大鼠腹主动脉球囊损伤后再狭窄中的作用,为防治血管再狭窄提供新的理论依据。方法选取雄性SD大鼠24只,随机分为假手术组(n=12)和球囊组(n=12)。在球囊组,将球囊导管自左颈总动脉插至腹主动脉末端扩张回抽,造成损伤;假手术组仅行颈部正中切开。两组均在术后30 d取材,常规组织切片观察血管病理学改变,RT-PCR法检测血管组织中GSK-3β和白细胞介素-2(IL-2)的mRNA表达变化,Western blot法检测CaN、活化T细胞核因子1(NFATC1)、GSK-3β、磷酸化GSK-3β(P-GSK-3β)在血管壁中的蛋白表达水平。结果球囊组损伤后血管壁内膜明显增殖,新生内膜厚度不均;球囊组较假手术组内膜/中膜厚度明显增加(P<0.01)。球囊组IL-2 mRNA表达量比假手术组表达升高(P<0.01),球囊组GSK-3βmRNA表达量与假手术组差异无统计学意义(P>0.05)。球囊组血管组织CaN和NFATC1、P-GSK-3β蛋白表达量均较假手术组明显升高(P<0.05)。结论球囊损伤后血管内膜增殖,伴随CaN-NFAT信号通路及其负调控信号GSK-3β途径的活化。

Objective To study the CaN -NFAT signaling pathway and the suppression factor glycogen synthase 3β （ GSK - 3β） in restenosis after percutaneous angioplasty （PTA） , and to investigate the restenosis mechanism of rat abdominal aorta balloon injury. Methods Twenty - four male SD rats were randomly divided into control group （ n = 12 ） and balloon injury group （n = 12）. In the balloon injury group, the balloon catheter was inserted from the left carotid to the end of the abdominal aorta to produce intima injury, neck midline incision alone was performed in control group. Then specimens were obtained after 30 days. HE staining was performed pathological observation of arterial wall. The real - time PCR was used to assessed the expression level of IL -2 and GSK -3β in arterial wall. CaN, NFATC1 and GSK, and P- GSK- 3β were determined by Western blot. Results Significant increase of intima/medium ratio was revealed in balloon injury group （P 〈0. 01 ）. There was significant up -regulation of IL- 2 in balloon injury group （P 〈 0. 05 ）, while there was no significant difference in the expression of GSK -3β between the two groups. Significant up -regulation of CaN, NFATC1 and P - GSK -3β was observed in balloon injury group according to Western blot （P 〈 0. 05 ）. Conclusion The intima is proliferated after balloon injury, accompanied with activation of CaN - NFAT signaling pathway as well as its negative regulation of signal GSK- 3β pathway.