血管紧张素(1-9)在内皮细胞中对血管紧张素Ⅱ诱导的血管细胞黏附分子1的调控研究

Role of Ang-(1-9)in regulating AngⅡ-induced expression of VCAM-1 in human vascular endothelial cells

目的研究血管紧张素(Ang)家族新成员Ang-(1-9)对动脉粥样硬化中关键黏附分子血管细胞黏附分子1(VCAM-1)表达的作用及调控机制。方法在体外培养人脐静脉内皮细胞,给予相应刺激进行分组:对照组;AngⅡ为实验1组;Ang-(1-9)为实验2组;AngⅡ+Ang-(1-9)为实验3组;AngⅡ+氯沙坦为实验4组;AngⅡ+Ang-(1-9)+PD123319为实验5组。采用RT-PCR及流式细胞仪检测VCAM-1 mRNA及蛋白表达,荧光素酶报告基因检测法测定VCAM-1启动子活性,酶联免疫吸附法测定NF-κB细胞核内水平,细胞免疫荧光观察NF-κB细胞核内转移情况。结果与对照组比较,实验1组VCAM-1 mRNA和蛋白表达明显增加(2.61±0.05 vs 1.00±0.00,146.35±1.81 vs 11.17±0.52,P<0.05);与实验1组比较,实验3组和实验4组VCAM-1 mRNA和蛋白表达明显减少,差异有统计学意义(P<0.05)。与对照组比较,实验1组VCAM-1启动子活性和NF-κB在细胞核内表达明显增加,差异有统计学意义(P<0.05);与实验1组比较,实验3组VCAM-1启动子活性和NF-κB在细胞核内表达明显降低,差异有统计学意义(P<0.05);与实验3组比较,实验5组VCAM-1启动子活性和NF-κB在细胞核内表达明显增加,差异有统计学意义(P<0.05)。结论Ang-(1-9)对VCAM-1的抑制作用有助于为动脉粥样硬化疾病的防治提供新思路。

Objective To study the effect of Ang-(1-9)on expression of VCAM-1 in atherosclerosis and its mechanism.Methods Human umbilical vein endothelial cells cultured in vitro were divided into control group,experimental group 1,experimental group 2,experimental group 3,experimental group 4 and experimental group 5.The expressions of VCAM-1 mRNA and protein were detected by RT-PCR and flow cytometry respectively.The activity of VCAM-1 promoter was assayed by luciferase report gene assay.The serum NF-κB level in nuclei was measured by ELISA.The transfer of NF-κB in nuclei was detected by cell immunofluorescence.Results The expression levels of VCMA-1 mRNA and protein were significantly higher in experimental group 1 than in control group(2.61±0.05 vs 1.00±0.00,146.35±1.81 vs 11.17±0.52,P<0.05)and significantly lower in experimental groups 3 and 4 than in experimental group 1(P<0.05).The activity of VCAM-1 promoter and expression level of NF-κB in nuclei were significantly higher in experimental group 1 than in control group(P<0.05),in experimental group 5 than in experimental group 3(P<0.05),and significantly lower in experimental group 3 than in experimental group 1(P<0.05).Conclusion The inhibitory effect of Ang-(1-9)on expression of VCAM-1 can provide new ideas for the prevention and treatment of atherosclerotic diseases.