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

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、１０００、２０００Ｕ／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２４ｈ,并以等量的０．０１ｍｏｌ／Ｌ、ｐＨ７．４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２０００Ｕ／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

Effects of tumor necrosis factor-alpha on proliferation and intracellular free calcium concentration in endothelium of human umbilical vein endothelial cells

OBJECTIVE: To Observe the effects of tumor necrosis factor-alpha (TNF-alpha) on cell proliferation and Intracellular free calcium concentration ([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-alpha 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 fluorospectrophotmetry. RESULTS: Incubated with TNF-alpha, 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-alpha. Stimulated by TNF-alpha, the percentage of cellcycles from phase G0 + G1 to S and G2 + M decreased significantly and it was dose-dependent [Ca2+]i increased significantly and dose-dependent as well. CONCLUSION: TNF-alpha may injure endothelium directly and inhibit its proliferation and repair through the changes of [Ca2+]i level. It may play an important role in the pathogenesis of PIH.