人工合成E选择素抑制大鼠血管平滑肌细胞增殖机制的初步研究  

Preliminary study of artificial synthetic E-selectin inhibits the proliferation mechanism of vascular smooth muscle cells in rats

作　　者：胡小辉[1] 刘建刚[1] 黄亚波[1] 吴江[1] 张世明[1] 

机构地区：[1]苏州大学附属第一医院神经外科,215006

出　　处：《中华神经外科杂志》2016年第5期506-512,共7页Chinese Journal of Neurosurgery

基　　金：国家自然科学基金（81371280）;苏州市科技应用基础研究项目（SYS201535）;苏州大学研究生创新工程项目（ZY32007213）

摘　　要：目的初步研究人工合成E选择素抑制大鼠血管平滑肌细胞增殖的机制。方法成年雄性SD大鼠90只，按随机数字表顺序分为对照组、手术组和治疗组，每组30只。手术组和治疗组行右侧颈总动脉纵行切开和间断缝合，对照组不行血管切开。缝合血管后，治疗组和手术组从股静脉分别注入人工合成E选择素和等量生理盐水。术后第1、3、7和14天采用酶联免疫吸附法测定血清肿瘤坏死因子OL（TNF-α）和白细胞介素6（IL-6）的浓度；取缝合口处血管组织，采用Western blot和实时定量（RT）-PCR检测NF—κB p65蛋白水平和mRNA的表达，凝胶电泳迁移率实验检测NF-KB转录活性，免疫组化法检测增殖细胞核抗原（PCNA）阳性的细胞率。结果手术组和治疗组血清中TNF-α和IL-6浓度、血管组织中NF—κB p65蛋白和mRNA水平均从缝合术后第1天开始升高，第3天达高峰，第7天时仍均高于对照组，差异有统计学意义（P〈0．01），而治疗组上述指标均低于同期手术组（P〈0．01），第14天3组间差异无统计学意义（P〉0．05）；缝合术后第3天，手术组NF—κB转录活性高于治疗组（P〈0．01）；手术组和治疗组PCNA阳性细胞从缝合术后第3天开始表达，对照组表达不明显或无表达，3组间差异有统计学意义（P〈0．05），第7、14天仍高表达（P〈0.01），而治疗组PCNA阳性细胞表达均低于同期手术组（P〈0．01）。结论人工合成E选择素可通过抑制NF—κB活性抑制血管平滑肌细胞的增殖。Objective To preliminary study the mechanism of artificial synthetic E-selectin inhibiting the proliferation of vascular smooth muscle cells in rats. Methods Ninety adult male SD rats were randomly divided into 3 groups ： a control group, an operation group, and a treatment group （ n = 30 in each group）. The right common carotid artery was made a longitudinal incision and interrupted suture in the operation group and the treatment group, and the control group did not make vascular incision and suture. Artificial synthetic E-selectin and the same amount of saline were injected into the femoral veins of the rats respectively in the treatment group and the operation group after vascular suture. On day 1, 3, 7, and 14 after procedure, ELISA was used to detect the concentrations of serum tumor necrosis factor-α（TNF-α） and intedeukin-6 （IL-6）. The vascular tissue at the suture was taken. Western blot and RT-PCR were used to detect the protein level of NF-κB p65 and mRNA expression. Eleetrophoretic mobility shift assay was used to detect the NF-κB transcriptional activity. Immunohistochemical staining was used to detect the positive cell rate of proliferating cell nuclear antigen （PCNA）. Results The concentrations of serum TNF-α and IL-6 and the levels of NF-κB p65 protein and mRNA began to increase at day 1 after vascular suture in the operation group and the treatment group. They reached the peak at day 3, and they were still higher than those in the control group at day 7 （P 〈 0.01 ）, and the above indices in the treatment group were lower than those in the operation group over the same period. There were significant differences （ P 〈 0.01 ） , and there were no significant differences among the 3 groups at day 14 （ P 〉 0. 05 ）. The NF-κB transcription activity in the operation group was higher than that in the treatment group at day 3 after suture （ P 〈 0.01 ）. The PCNA positive cells began to express at day 3 after vascular suture in the operation group and the treatm

关 键 词：E选择素 NF—κB 肌细胞 平滑肌 细胞增殖 大鼠 血管 

分 类 号：R653[医药卫生—外科学]