Apoptosis in vascular endothelial cells caused by serum deprivation, oxidative stress and transforming growth factor-β
Apoptosis in vascular endothelial cells caused by serum deprivation, oxidative stress and transforming growth factor-β
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DOI：
10.3109/10623329909165310
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发表时间：
1999-01-01
影响因子：
--
通讯作者：
Gobé, G
中科院分区：
文献类型：
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作者：
Hogg, N;Browning, J;Gobé, G
血管内皮细胞凋亡已被证明在高血压诱导的血管缺失和损伤的发病机制中发挥作用。在目前的体外研究中，我们分析了几种可能的相关致病因素的血管内皮细胞凋亡，即，血清剥夺和营养耗竭，氧化应激的形式缺氧，高氧或自由基损伤，和改变水平的转化生长因子-β 1（TGF-β 1）蛋白。将已建立的细胞系牛主动脉内皮细胞（BAEC）保存在25 cm（2）烧瓶或12孔板盖玻片上的完全生长培养基（RPMI-1640加15%胎牛血清和抗生素，缩写为RPMI）中。将汇合但活跃生长的培养物用缺氧（RPMI的PO 2 = 50 mmHg）、无血清培养基（SFM）、SFM加缺氧、高氧（RPMI的PO 2 = 450 mmHg）、SFM中的过氧化氢（H2 O2，1 mM）或SFM中的TGF-β 1蛋白（10 ng/mL）处理。使用适当的对照培养物。除TGF-β_1和H_2O_2处理后16- 18 h收集BAEC外，其余处理后48 h或72 h收集BAEC。使用形态学特征或原位末端标记（ISEL）评估细胞死亡，使用增殖细胞核抗原（PCNA）评估细胞增殖，使用转录水平或免疫组织化学评估TGF-β 1表达。与对照组相比，所有处理组的细胞凋亡水平均显著增加（P
Vascular endothelial cell apoptosis has previously been shown to play a role in the pathogenesis of hypertension-induced vessel deletion and damage. In the present in vitro study we analyse several possible relevant causative factors of vascular endothelial cell apoptosis, namely, serum deprivation and nutrient depletion, oxidative stress in the forms of hypoxia, hyperoxia or free radical damage, and altered levels of transforming growth factor-beta 1 (TGF-beta 1) protein. An established cell line, bovine aortic endothelial cells (BAEC), was maintained in complete growth medium (RPMI-1640 plus 15% fetal calf serum and antibiotics, abbreviated as RPMI) in 25cm(2) flasks or in 12-well plates on glass coverslips. Confluent but actively-growing cultures were treated with either hypoxia (PO2 of RPMI = 50mmHg), serum-free media (SFM), SFM plus hypoxia, hyperoxia (PO2 of RPMI = 450mmHg), hydrogen peroxide (H2O2, 1 mM) in SFM, or TGF-beta 1 protein (10ng/mL) in SFM. Appropriate control Cultures were used. BAEC were collected 48h or 72h after all treatments except for TGF-beta 1 and H2O2 treatments that were collected at 16-18h. Cell death was assessed using morphological characteristics or in situ end labeling (ISEL), cell proliferation assessed using proliferating cell nuclear antigen (PCNA), and TGF-beta 1 expression assessed using transcript levels or immunohistochemistry. All treatments significantly increased levels of apoptosis over control cultures (P