Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral St., Glasgow G4 0RE, Scotland, United Kingdom.
Toxicol In Vitro. 2011 Dec;25(8):1680-6. doi: 10.1016/j.tiv.2011.07.005. Epub 2011 Jul 18.
Peroxynitrite is widely reported as highly cytotoxic; yet recent evidence indicates that at certain concentrations, it can induce pulmonary cell hyper-proliferation and tissue remodelling. This study aimed to establish the threshold concentration of peroxynitrite to induce functional impairment of bovine pulmonary artery endothelial (PAEC) and smooth muscle cells (PASMC). PAEC or PASMC were exposed to solution of peroxynitrite or 3-morpholinosydnonimine (SIN-1). Twenty-four hour cell viability, DNA synthesis, and protein biochemistry were assessed by trypan blue dye exclusion, [3H] thymidine incorporation and western blot analysis, respectively. Threshold concentration of peroxynitrite to significantly impair viability of PAEC and PASMC was 2 μM peroxynitrite. In PASMC and PAEC, low concentrations of peroxynitrite (2 nM-0.2 μM) increased cell proliferation and did not activate p38 MAP kinase. The decrease in DNA synthesis and cell viability caused by 2 μM peroxynitrite was associated with caspase-3 cleavage but not p38 activation. Also, 2-20 μM peroxynitrite significantly activated poly ADP ribose polymerase and stress activated kinase JNK in PAEC. However, the higher concentration of 20 μM peroxynitrite did cause a threefold increase in p38 activation. In conclusion, the threshold for the cytotoxic effects of peroxynitrite was 2 μM; which caused apoptotic cell death independent of p38 MAP kinase activation in pulmonary artery cells.
过氧亚硝酸盐被广泛报道为具有高度细胞毒性；然而，最近的证据表明，在特定浓度下，它可以诱导肺细胞过度增殖和组织重塑。本研究旨在确定诱导牛肺动脉内皮细胞（PAEC）和肺动脉平滑肌细胞（PASMC）功能障碍的过氧亚硝酸盐的阈值浓度。将 PAEC 或 PASMC 暴露于过氧亚硝酸盐或 3-吗啉代-sydnonimine（SIN-1）溶液中。通过台盼蓝染料排除法、[3H]胸苷掺入法和 Western blot 分析分别评估 24 小时细胞活力、DNA 合成和蛋白质生化。显著损害 PAEC 和 PASMC 活力的过氧亚硝酸盐阈值浓度为 2 μM 过氧亚硝酸盐。在 PASMC 和 PAEC 中，低浓度的过氧亚硝酸盐（2 nM-0.2 μM）增加细胞增殖，并且不激活 p38 MAP 激酶。2 μM 过氧亚硝酸盐引起的 DNA 合成和细胞活力下降与 caspase-3 切割有关，但与 p38 激活无关。此外，2-20 μM 过氧亚硝酸盐在 PAEC 中显著激活聚 ADP 核糖聚合酶和应激激活激酶 JNK。然而，更高浓度的 20 μM 过氧亚硝酸盐确实导致 p38 激活增加了三倍。总之，过氧亚硝酸盐细胞毒性的阈值为 2 μM；这导致了肺动脉细胞中独立于 p38 MAP 激酶激活的凋亡性细胞死亡。
