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肿瘤坏死因子α对脐静脉血管内皮细胞增殖和细胞内游离钙浓度的影响

更新时间:2012-04-21 13:23:23  
金镇 裴凌 王德智

  【摘要】 目的 探讨肿瘤坏死因子α(TNF-α)对培养的人脐静脉血管内皮细胞增殖和细胞内游离钙浓度的影响,从细胞间信息传递水平探讨妊娠高血压综合征(妊高征)的发病机理。方法 用终浓度为500、1 000、2 000 U/ml的TNF-α,将融合成单层的人脐静脉血管内皮细胞进行无血清培养24 h,并以等量的0.01 mol/L、pH 7.4的磷酸盐缓冲生理盐水作对照,用流式细胞仪测定细胞各周期细胞百分数、荧光分光光度法测定内皮细胞内游离钙浓度。结果 经TNF-α作用后的内皮细胞被拉长,由原来的上皮型变为成纤维细胞型,细胞及核边界尚清楚、形态较规则,但在2 000 U/ml浓度时,可见有细胞脱落及细胞内异常颗粒出现。内皮细胞受到TNF-α的刺激后,由G0+G1期进入S和G2+M期的细胞百分比明显减少,G0+G1期细胞百分数增多,S和G2+M期细胞明显减少,并呈剂量依赖性,与对照组比较,差异有非常显著性。细胞内游离钙浓度也明显升高,并呈剂量依赖性,与对照组比较,差异也有非常显著性。结论 TNF-α可直接引起内皮细胞损伤,并可抑制内皮细胞的增殖和损伤后的修复。TNF-α损伤内皮细胞的作用是以钙离子为信使进行细胞间信息传递的,对妊高征的发病起到重要的作用。
  【关键词】 肿瘤坏死因子; 内皮,血管; 细胞分裂; 钙

Effects of Tumor Necrosis Factor-α on Proliferation and Intracellular Free Calcium Concentration in Endothelium of Human Umbilical Vein Endothelial Cells

JIN Zhen*, PEI Ling, WANG Dezhi
(Department of Obstetric and Gynecology, The Second Hospital, China Medical University, Shenyang 110003, China)

  【Abstract】 Objective To Observe the effects of tumor necrosis factor-α (TNF-α) on cell proliferation and Intracellular free calcium concentraction ([Ca2+]i) in endothelium of human umbilical vein endothelial cells (HUVEC) and investigate the pathogenesis of pregnancy induced hypertension syndrome (PIH). Methods Confluent monolayer of HUVEC was directly incubated with TNF-α at following final concentrations: 500, 1 000, 2 000 U/ml for 24 hours. The percentages of different cellcycles and [Ca2+]i were measured by flow cytometry and fluorospectrophotometry. Results Incubated with TNF-α, the endothelial cells were elongated and transformed into fibroblast-like cells. Border of nucleus was sharp, clarity, and cells were in regular shape. But there were abnormal granules in cytoplasma and some cells detached at the concentrotion of 2 000 U/ml of TNF-α. Stimulated by TNF-α, the percentage of cellcycles from phase G0+G1 to S and G2+M decreased significantly

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