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目的探讨抑肽酶对肿瘤坏死因子-α(TNF-α)活化人脐静脉内皮细胞的作用。方法采用Ficoll液提取高纯度的健康成人中性粒细胞(PMN)。体外培养的人脐静脉内皮细胞随机分为3 组,对照组(C组):给予等量培养液;TNF-α活化组(T组):给予TNF-α 2 ng/ml,作用时间5 h;抑肽酶+ TNF-α组(A+T组):给予抑肽酶250 KIU／ml,作用时间6 h,1 h后给予TNF-α2 ng/ml,作用时间5 h。用细胞培养小室测定PMN跨迁单层融合的人脐静脉内皮细胞的迁移率,采用免疫细胞化学法测定人脐静脉内皮细胞细胞间黏附分子-1(ICAM-1)以及细胞连接蛋白β-链蛋白的表达。结果与C组比较, T A+T组ICAM-1表达和PMN迁移率增加,β-链蛋白表达下降(P<0．05);与T组比较,A+T组 ICAM-1表达和PMN迁移率降低,β-链蛋白表达增多(P<0．05)。结论抑肽酶可抑制TNF-α对人脐静脉内皮细胞的活化。
Objective To investigate the effect of aprotinin on the activation of tumor necrosis factor-α (TNF-α) in human umbilical vein endothelial cells. Methods Ficoll liquid was used to extract healthy adult neutrophils (PMN) with high purity. Human umbilical vein endothelial cells cultured in vitro were randomly divided into 3 groups: control group (C group): equal volume of culture medium; TNF-α activation group (T group): given TNF-α 2 ng / ; Aprotinin + TNF-α group (A + T group): given aprotinin 250 KIU / ml, the role of time 6 h, 1 h after the given TNF-α2 ng / ml, the role of time 5 h. The migration of monolayer-fused human umbilical vein endothelial cells (PMNs) was measured by cell culture chamber. The expression of ICAM-1 and beta-catenin in human umbilical vein endothelial cells were detected by immunocytochemistry Protein expression. Results Compared with group C, the expression of ICAM-1 and PMN increased and the expression of β-catenin decreased in T A + T group (P <0.05). Compared with group T, the expression of ICAM-1 and PMN The migration rate decreased and the expression of β-catenin increased (P <0.05). Conclusion Aprotinin inhibits the activation of human umbilical vein endothelial cells by TNF-α.