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目的探讨细胞周期素依赖性激酶抑制蛋白P27在大鼠胸主动脉球囊损伤后管腔狭窄过程中的表达变化规律及全反式维甲酸对其的影响。方法将54只400～500g的雄性SD大鼠随机分为三组,全反式维甲酸治疗组大鼠行胸主动脉球囊损伤术,术前4天开始予全反式维甲酸(30mgkg.d)灌胃至术后处死;单纯手术组大鼠行胸主动脉球囊损伤术;对照组不行球囊损伤,作为正常对照;手术组和对照组予对照用芝麻油灌胃。分别于术后2、7、14、28天取胸主动脉用HE染色、免疫组织化学和计算机图像分析仪进行形态学、增殖细胞核抗原和P27表达水平检测。结果①正常动脉壁不表达增殖细胞核抗原,球囊损伤后开始表达,术后2天中膜表达显著(24.08±2.35);术后7天新生内膜高度表达增殖细胞核抗原(35.32±3.46),而中膜表达明显下降(9.47±1.56);后内膜、中膜增殖细胞核抗原表达均逐渐下降,28天时形成显著的新生内膜(0.173±0.030mm2),管腔狭窄(1.641±0.088mm2)。②正常动脉壁显著表达P27(19.29±1.54),损伤后中膜表达迅速下降,2天时达最低水平(2.93±0.55),后逐渐回升;14天、28天时新生内膜中可见P27表达,并逐渐增多(14天:10.30±1.39;28天:16.01±1.33)。③P27表达与增殖细胞核抗原表达呈显著负相关(r=-0.868,P<0.001)。④全反式维甲酸治疗后明显抑制中膜P27的下调(2天:7.67±1.27),促进新生内膜中P27表达(7天:6.09±1.04;14天:22.60±2.46;28天:26.23±2.43),同时显著抑制增殖细胞核抗原的表达(2天时中膜:12.11±1.84;7天时内膜:16.83±1.02),新生内膜面积明显减少(0.075±0.017mm2),管腔面积(1.901±0.085mm2)明显大于单纯手术组(P均<0.01)。结论P27低表达在动脉损伤后管腔狭窄过程中具有重要意义。
Objective To investigate the expression of cyclin-dependent kinase inhibitor P27 in rat stenosis after thoracic aorta balloon injury and the effect of all-trans retinoic acid on it. Methods 54 male SD rats (400-500g) were randomly divided into three groups. All-trans retinoic acid-treated rats underwent thoracic aortic balloon injury. All-trans retinoic acid (30mgkg. d) Gavage to sacrifice after operation; Thoracic aorta balloon injury in rats in operation group; Balloon injury in control group as normal control; Operation and control groups were given gavage with sesame oil. The thoracic aorta was taken at 2, 7, 14, and 28 days after operation. HE staining, immunohistochemistry and computer image analyzer were used to detect the morphology, proliferating cell nuclear antigen and P27 expression. Results ① The proliferating cell nuclear antigen was not expressed in the normal artery wall, and then began to be expressed after balloon injury. The expression of the proliferating cell nuclear antigen was significantly higher in the neointima at 2 days after operation (24.08 ± 2.35) (35.32 ± 3.46) (9.47 ± 1.56). The expressions of proliferating cell nuclear antigen (PCNA) in the intima and media gradually decreased after 28 days, and significant neointimal formation (0.173 ± 0.030mm2) and stenosis (1.641 ± 0.088mm2) . P27 (19.29 ± 1.54) was significantly increased in normal artery wall, the expression of P27 was rapidly decreased after injury, reached the lowest level at 2 days (2.93 ± 0.55), and then gradually rose. P27 expression was seen in neointima at 14 and 28 days Increasingly (14 days: 10.30 ± 1.39; 28 days: 16.01 ± 1.33). P27 expression was significantly negatively correlated with PCNA expression (r = -0.868, P <0.001). ④ All-trans retinoic acid treatment significantly inhibited the down-regulation of P27 (2 days: 7.67 ± 1.27), and promoted the expression of P27 in neointima (7 days: 6.09 ± 1.04; 14 days: 22.60 ± 2.46; 28 days: 26.23 ± 2.43), and significantly inhibited the expression of proliferating cell nuclear antigen (12.11 ± 1.84 at 2 days; 16.83 ± 1.02 at 7 days), neointimal area was significantly reduced (0.075 ± 0.017mm2), lumen area (1.901 ± 0.085mm2) was significantly greater than the simple operation group (P <0.01). Conclusion The low expression of P27 is of great significance in the process of stenosis after arterial injury.