Elevated fibroblast growth factor-2 increases tumor necrosis factor-α, induced endothelial cell death in high glucose
Elevated fibroblast growth factor-2 increases tumor necrosis factor-α, induced endothelial cell death in high glucose
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DOI：
10.1002/jcp.21476
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发表时间：
2008-10-01
影响因子：
5.6
通讯作者：
Edelman, Elazer R.
中科院分区：
文献类型：
--
作者：
Clyne, Alisa Morss;Zhu, Han;Edelman, Elazer R.
糖尿病患者血糖和肿瘤坏死因子-α（TNF α）浓度升高。这两个因素与糖尿病血管病变和内皮细胞凋亡相关，但它们的联合作用尚未被测量。我们以前已经表明，血管生成生长因子成纤维细胞生长因子-2（FGF-2），这是一般保护内皮细胞死亡，类似地在高葡萄糖条件下升高。因此，我们研究了在正常和升高的葡萄糖条件下TNF α对内皮细胞死亡的影响，特别关注FGF-2。将猪主动脉内皮细胞在5和30 mM葡萄糖中培养，并用TNF α与FGF-2或中和FGF-2抗体一起刺激。通过细胞计数或膜联蛋白凋亡测定来测量细胞死亡，并通过碘化丙啶标记来确定细胞周期时相。TNF α诱导的内皮细胞死亡增加的细胞在高葡萄糖，和细胞死亡增强与FGF-2暴露的增加和否定的中和FGF-2抗体。当在TNF α之前18 h用FGF-2刺激时，内皮细胞对TNF α诱导的细胞死亡最敏感，对应于细胞进入增殖周期的S期。FGF-2相关的TNF α诱导的细胞死亡的增加被否定的阻断细胞进入S期。在高糖环境中，内皮细胞释放FGF-2导致细胞周期进展，这使得细胞对TNF α诱导的细胞死亡更敏感。这些数据表明，生长因子在高葡萄糖的结果取决于二级介质，如细胞因子和刺激细胞周期的时间。
Glucose and tumor necrosis factor-alpha (TNF alpha) concentrations are elevated in diabetes. Both of these factors correlate with diabetic vasculopathy and endothelial cell apoptosis, yet their combined effects have not been measured. We have previously shown that the angiogenic growth factor fibroblast growth factor-2 (FGF-2), which is generally protective against endothelial cell death, is similarly elevated in high glucose conditions. We therefore investigated the effect of TNFa on endothelial cell death under normal and elevated glucose conditions, with a particular focus on FGF-2. Porcine aortic endothelial cells were cultured in 5 and 30 mM glucose and stimulated with TNF alpha, together with FGF-2 or a neutralizing FGF-2 antibody. Cell death was measured via cell counts or an annexin apoptotic assay, and cell cycle phase was determined by propidium iodide labeling. TNF alpha-induced endothelial cell death increased for cells in high glucose, and cell death was enhanced with increasing FGF-2 exposure and negated by a neutralizing FGF-2 antibody. Endothelial cells were most susceptible to TNFa-induced cell death when stimulated with FGF-2 18 h prior to TNFa, corresponding to cell entry into S phase of the proliferative cycle. The FGF-2 associated increase in TNFa-induced cell death was negated by blocking cell entry into S phase. Endothelial cell release of FGF-2 in high glucose leads to cell cycle progression, which makes cells more susceptible to TNF alpha-induced cell death. These data suggest that growth factor outcomes in high glucose depend on secondary mediators such as cytokines and stimulation cell cycle timing.