Protection against peroxynitrite-induced fibroblast injury and arthritis development by inhibition of poly(ADP-ribose) synthase
Protection against peroxynitrite-induced fibroblast injury and arthritis development by inhibition of poly(ADP-ribose) synthase
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DOI：
10.1073/pnas.95.7.3867
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发表时间：
1998-03-31
影响因子：
11.1
通讯作者：
Kun, E
中科院分区：
文献类型：
--
作者：
Szabó, C;Virág, L;Kun, E
过氧亚硝酸盐，一种由一氧化氮（NO）和超氧化物形成的细胞毒性氧化剂，诱导DNA链断裂，其激活核酶聚（ADP-核糖）合酶（PARS; EC 2.4.2.30）。out动物（PARS（-/-））和相应的野生型动物（PARS（+/+）），借助亲脂性PARS抑制剂5-碘-6-氨基-1，2-苯并吡喃酮（INH 2BP）。我们研究了PARS在体外过氧亚硝酸盐诱导的成纤维细胞损伤中的作用，以及在体内关节炎的发展中的作用。PARS（+/+）动物的胚胎成纤维细胞暴露于过氧亚硝酸盐引起DNA单链断裂和PARS活化，并引起线粒体呼吸的急性抑制。INH 2BP保护PARS（+/+）细胞免受过氧亚硝酸盐（50-100 μ M）引起的线粒体呼吸抑制。与INH 2BP抑制PARS类似，PARS（-/-）细胞对过氧亚硝酸盐诱导的损伤具有保护作用，当细胞被高浓度氧化剂攻击时，PARS（-/-）表型或INH 2BP对细胞损伤的保护作用减弱，INH 2BP或PARS（-/-）表型抑制PARS降低诱导型一氧化氮合酶（iNOS; EC 1.14.13.39）mRNA水平并抑制NO的产生。在免疫刺激的细胞中，INH 2BP没有清除过氧亚硝基阴离子或羟基自由基的作用，并且它没有额外的在PARS（-/-）细胞中的（非特异性）作用，在胶原诱导的关节炎中，在发炎的关节中发现了硝基酪氨酸（过氧亚硝酸盐形成的标志物）的显著染色，（0.5g/kg，每日一次），在关节炎发作时（第25天）开始，延迟了第26-35天临床体征的发展，并改善了膝关节和爪的组织学状态，我们的数据表明，通过基因操作或药物抑制PARS来删除PARS可以保护机体免受氧化-在体外诱导细胞损伤并在体内表现出抗炎作用。
Peroxynitrite, a cytotoxic oxidant formed from nitric oxide (NO) and superoxide, induces DNA strand breakage, which activates the nuclear enzyme poly(ADP-ribose) synthase (PARS; EC 2.4.2.30), The cellular function of PARS was determined in fibroblast lines from PARS knock; out animals (PARS(-/-)) and corresponding wild type animals (PARS(+/+)), with the aid of the lipophilic PARS inhibitor 5-iodo-6-amino-1,2-benzopyrone (INH2BP). We investigated the role of PARS in peroxynitrite-induced fibroblast injury in vitro and also in the development of arthritis in vivo. Exposure of embryonic fibroblasts from the PARS(+/+) animals to peroxynitrite caused DNA single-stand breakage and PARS activation and caused an acute suppression of mitochondrial respiration, INH2BP protected the PARS(+/+) cells against the suppression of mitochondrial respiration in response to peroxynitrite (50-100 mu M). Similarly to PARS inhibition with INH2BP, the PARS(-/-) cells were protected against peroxynitrite-induced injury, The protection against cellular injury by PARS(-/-) phenotype or INH2BP waned when cells were challenged with higher concentrations of the oxidant, Inhibition of PARS by INH2BP or by PARS(-/-) phenotype reduced inducible nitric-oxide synthase (iNOS; EC 1.14.13.39) mRNA levels and inhibited production of NO in immunostimulated cells, INH2BP had no peroxynitrite scavenging or hydroxyl radical scavenging effects, and it exerted no additional (nonspecific) effects in the PARS(-/-) cells, In collagen-induced arthritis, significant staining for nitrotyrosine, a marker of peroxynitrite formation, was found in the inflamed joints, Oral treat ment with INH2BP (0.5 g/kg, daily), starting at the onset of arthritis (day 25), delayed the development of the clinical signs at days 26-35 and improved histological status in the knee and paw, Our data demonstrate that deletion of PARS by genetic manipulation or pharmacological inhibition of PARS protects against oxidant-induced cellular injury in vitro and exhibits anti-inflammatory effects in vivo.