纤维蛋白(原)及其降解产物对共培养人脐静脉内皮细胞的细胞间黏附分子-1表达的影响

Effect of fibrinogen and its degradation products on the expression of intercellular adhesion molecule-1 in coculture system of human umbilical vein endothelial cells

目的探讨纤维蛋白原(Fg)、纤维蛋白(Fb)及其降解产物(FDPs)对人脐静脉内皮细胞(HU-VECs)/兔平滑肌细胞(SMCs)共培养模型中细胞间黏附分子-1(ICAM-1)表达的影响。方法建立原代HU-VECS与兔SMC共培养体系,根据干预物及其浓度的不同分为Fg、Fb、FDPs组及相应的0 mg/ml、0.5 mg/ml、3.0 mg/ml和6.0 mg/ml亚组。应用核因子B抑制蛋白(I-B)拮抗剂BAY 11-7082、蛋白激酶C(PKC)拮抗剂Staurosporine分别对0.5 mg/ml、3.0 mg/ml和6.0 mg/ml亚组进行干预。采用反转录(RT)-PCR法检测HU-VECS内ICAM-1 mRNA水平;酶联免疫吸附法(ELISA)检测培养上清液的ICAM-1蛋白含量。结果 Fg 3.0mg/ml和6.0 mg/ml亚组ICAM-1 mRNA水平和蛋白含量显著高于0 mg/ml亚组及相应浓度Fg+BAY 11-7082亚组(均P<0.05);Fg 3.0 mg/ml亚组ICAM-1 mRNA水平和蛋白含量显著高于3.0 mg/ml+Staurosporine亚组(均P<0.05)。Fb 6.0 mg/ml亚组ICAM-1 mRNA水平显著高于0 mg/ml、6.0 mg/ml+BAY11-7082和6.0 mg/ml+Staurosporine亚组(均P<0.05),但Fb 6.0 mg/ml亚组ICAM-1蛋白含量显著高于0 mg/ml亚组(P<0.05)。FDPs 6.0 mg/ml亚组ICAM-1 mRNA水平和蛋白含量显著高于0 mg/ml、相应浓度BAY11-7082和Staurosporine亚组(均P<0.05)。结论中、高浓度Fg、Fb、FDPs能上调血管内皮细胞ICAM-1的表达。

Objective To investigate the effect of fibrinogen （Fg） and its degradation products （FDPs） on the expression of intercellular adhesion molecule-1 （ ICAM-1 ） in coculture system of human umbilical vein endothelial ceils （HUVECs） and rabbit smooth muscle cells （SMCs）. Methods The coculture system of HUVECs in primary culture and rabbit SMCs was established. According to thedifferent interventions and concentrations, they were divided into Fg, Fb and FDPs groups and corresponding 0 mg/ml, O. 5 mg/ml, 3.0 mg/ml , 6.0 mg mg/ml subgroups. The each 3.0 mg/ml and 6.0 mg mg/ml subgroups were intervented by antagonist of inhibitory of nuclear factor B （I-B） BAY 11-7082 and antagonist of protein kinase C（PKC） Staurosporine. The level of ICAM-1 mRNA in HUVECs was detected by reverse transcriptase （RT）-PCR. ICAM-1 protein content from culture supernatant was detected by enzyme-linked immunosorbent assay （ELISA）. Results The levels of ICAM-1 mRNA and protein in Fg 3.0 mg/ml and 6.0 mg/ml subgroups were significantly higher than those in 0 mg/ml subgroup and corresponding concentrations Fg ＋ BAY 11-7082 subgroup （ all P 〈 0. 05 ） ; and the levels of ICAM-1 mRNA and protein in Fg 3.0 mg/ml subgroup were significantly higher than those in 3.0 mg/ml ＋ Staurosporine subgroup （ all P 〈 0. 05 ）. The levels of ICAM-1 mRNA in Fb 6.0 mg/ml subgroup was significantly higher than those in 0 mg/ml subgroup, 6.0 mg/ml ＋ BAY11-7082 and 6.0 mg/ml ＋ Staurosporine subgroups （ all P 〈 0. 05 ）. The content of ICAM-1 protein in Fb 6.0 mg./ml subgroup was significantly higher than that in 0 mg/ml subgroup （ P 〈 O. 05 ）. The levels of ICAM-1 mRNA and proteinin FDPs 6. 0 mg/ml subgroup were significantly higher than those in 0 mg/ml subgroup, corresponding concentrations FDPs ＋ BAY 11-7082 subgroup and Staurospofine subgroup （all P 〈 0. 05 ）. Conclusion The medium, high concentrations of Fg, Fb and FDPs can up-regulate the expression of ICAM-1 in vascular endothelial cell.