Serum from patients with severe heart failure downregulates eNOS and is proapoptotic -: Role of tumor necrosis factor-α
Serum from patients with severe heart failure downregulates eNOS and is proapoptotic -: Role of tumor necrosis factor-α
复制标题
DOI：
10.1161/01.cir.100.19.1983
复制
发表时间：
1999-11-09
期刊：
影响因子：
37.8
通讯作者：
Ferrari, R
中科院分区：
文献类型：
--
作者：
Agnoletti, L;Curello, S;Ferrari, R
背景-细胞因子激活和内皮功能障碍是充血性心力衰竭（CHF）的典型现象。我们测试了这样的假设：用 CHF 患者的血清孵育人脐静脉内皮细胞会下调内皮组成型一氧化氮合酶 (eNOS) 并诱导细胞凋亡。方法和结果 - 我们研究了 21 名严重 CHF 患者，评估了肿瘤坏死因子-α (TNF-α) 水平和一些神经内分泌参数。 eNOS 通过蛋白质印迹分析测量，细胞凋亡通过光学显微镜和流式细胞术测量。我们观察到(1) eNOS 下调（与健康受试者在 24 小时 [P < 0.05] 和 48 小时 [P < 0.001] 时的差异），(2) 细胞凋亡典型的核形态变化； (3) 碘化丙啶（48 小时从 2.1 +/- 0.4% 增加到 11.3 +/- 1.2%；与健康受试者相比，P < 0.001）和膜联蛋白 V 具有高细胞凋亡率。抗人 TNF-α 抗体不能完全抵消这些作用。 eNOS 下调与细胞凋亡之间存在很强的相关性（r = -8.89；P < 0.001）。结论：严重 CHF 患者的血清下调 eNOS 表达并增加细胞凋亡，高水平的 TNF-α 可能发挥作用，但不是唯一的因素。
Background-Cytokine activation and endothelial dysfunction are typical phenomena of congestive heart failure (CHF). We tested the hypothesis that incubating human umbilical vein endothelial cells with serum from patients with CHF will downregulate endothelial constitutive nitric oxide synthase (eNOS) and induce apoptosis,Methods and Results-We studied 21 patients with severe CHF, Levels of tumor necrosis factor-alpha (TNF-alpha) and several neuroendocrine parameters were assessed. eNOS was measured by Western Blot analysis and apoptosis by optical microscopy and flow cytometry. We observed(1) eNOS downregulation (difference versus healthy subjects at 24 hours [P < 0.05] and 48 hours [P < 0.001]), (2) nuclear morphological changes typical of apoptosis; and (3) a high apoptotic rate with propidium iodide (increasing from 2.1 +/- 0.4% to 11.3 +/- 1.2% at 48 hours; P < 0.001 versus healthy subjects) and annexin V. An anti-human TNF-alpha antibody did not completely counteract these effects. A strong correlation existed between eNOS downregulation and apoptosis (r = -8.89; P < 0.001).Conclusions-Serum from patients with severe CHF downregulates eNOS expression and increases apoptosis, High levels of TNF-alpha likely play a role, but they cannot be the only factor responsible.