Acrolein produces nitric oxide through the elevation of intracellular calcium levels to induce apoptosis in human umbilical vein endothelial cells: Implications for smoke angiopathy
Acrolein produces nitric oxide through the elevation of intracellular calcium levels to induce apoptosis in human umbilical vein endothelial cells: Implications for smoke angiopathy
复制标题
DOI：
10.1016/j.niox.2005.09.004
复制
发表时间：
2006-03-01
影响因子：
3.9
通讯作者：
Taniguchi, N
中科院分区：
文献类型：
--
作者：
Misonou, Y;Asahi, M;Taniguchi, N
丙烯醛是一种高度亲电子的α，β-不饱和醛，其水平在吸烟者的血液中增加。为了确定丙烯醛是否参与烟雾血管病的病理学，检查了丙烯醛对人脐静脉内皮细胞（HUVEC）的作用。细胞内一氧化氮（NO）的水平，测定使用二氨基荧光素-2二乙酸酯（DA-2 DA），一个NO敏感的荧光染料，被发现后，在HUVEC与10 μ M丙烯醛治疗增加。用2，3-二氨基萘测量亚硝酸盐和Western印迹分析显示，亚硝酸盐和S-亚硝基半胱氨酸水平以剂量依赖性方式增加，证实丙烯醛增加NO产生。用10 mM N-乙酰基-L-半胱氨酸（NAC）（一种抗氧化剂）处理不能降低这种增加，但用10 μ M细胞内钙螯合剂1，2-双（邻氨基苯氧基）乙烷-N，N，N ′，N ′-四乙酸四（乙酰氧基甲基）酯可降低这种增加。用1 mM NO合成酶抑制剂N（G）-硝基-L-精氨酸甲酯（L-NAME）预处理可显著降低丙烯醛刺激的NO生成。丙烯醛的细胞毒性降低预处理与10 μ M 2-（4-羧基苯基）-4，4，5，5-四甲基咪唑啉-1-氧基3-氧化物（羧基-PTIO），细胞内NO清除剂，或I MM LNAME，而不是减少10 mM NAC，20 μ M姜黄素，另一种过氧化物清除剂，或100 μ M Mn（III）TMPyP，超氧化物歧化酶模拟物。核染色和蛋白质印迹分析，使用抗切割的半胱天冬酶3抗体显示，减少的活力的HUVEC丙烯醛是由于细胞凋亡，这是逆转后预处理与0.1 mM羧基-PTIO或1 MM L-NAME。因此，丙烯醛增加细胞内钙的产生，诱导细胞内NO的产生，通过钙依赖性NO合成酶，可能eNOS，和NO的过度和快速增加可能导致HUVEC的凋亡。提示丙烯醛可能通过NO诱导内皮细胞凋亡参与烟雾性血管病的病理过程。(c)2005年爱思唯尔公司All rights reserved.
Acrolein is a highly electrophilic alpha, beta-unsaturated aldehyde, the levels of which are increased in the blood of smokers. To determine if acrolein is involved in the pathology of smoke angiopathy, the effect of acrolein on human umbilical vein endothelial cells (HUVEC) was examined. Intracellular nitric oxide (NO) levels, determined using diaminofluorescein-2 diacetate (DAF-2 DA), an NO sensitive fluorescent dye, were found to be increased after treatment in HUVEC with 10 mu M acrolein. The measurement of nitrite with 2,3-diaminonaphthalene and a Western blot analysis revealed that nitrite and S-nitroso-cysteine levels were increased in a dose-dependent manner, confirming that NO production is increased by acrolein. The increase was not reduced by treatment with 10 mM N-acetyl-L-cysteine (NAC), an anti- oxidant, but was reduced with 10 mu M of the intracellular calcium chelator, 1,2-bis (o-aminophenoxy) ethane-N,N,N',N'-tetraacetic acid tetra (acetoxymethyl) ester. Acrolein-stimulateds NO production was significantly reduced by pretreatment with I mM N(G)-nitro-L-arginine-methyl ester (L-NAME), an NO synthase inhibitor. The cytotoxicity of acrolein was reduced by pretreatment with 10 mu M 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (carboxy-PTIO), an intracellular NO scavenger, or I MM LNAME, whereas it was not reduced by 10 mM NAC, 20 mu M Curcumin, another peroxide scavenger, or 100 mu M Mn(III)TMPyP, a superoxide dismutase mimic. Nuclear staining and a Western blot analysis using an anti-cleaved caspase 3 antibody revealed that the reduced viability of HUVEC by acrolein was due to apoptosis, which was reversed after pretreatment with 0.1 mM carboxy-PTIO or 1 MM L-NAME. Thus, acrolein increases intracellular calcium production to induce intracellular NO production by a calcium-dependent NO synthase, possibly eNOS, and the excess and rapid increase in NO might lead to the apoptosis of HUVEC. These data suggest that acrolein might be involved in the pathology of smoke angiopathy through the NO-induced apoptosis of endothelial cells. (c) 2005 Elsevier Inc. All rights reserved.