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《Acta Physiological Sinica》 2003-04
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Endogenous peroxynitrite mediates lipopolysaccharide-induced injury in cultured pulmonary artery endothelial cells

GU Zhen-Yong~1,2, LING Yi-Ling~2, XU Xiao-Hu~1, ZHU Tie-Nian~2 , CONG Bin~2 1 Shantou University Medical College, Shantou 515031; 2 Department of Pathophysiology, Hebei Medical University, Shijiazhuang 050017  
This study, using cultured bovine pulmonary artery endothelial cells (BPAECs), was undertaken to investigate the roles of endogenous ONOO~- in LPS-caused injury in endothelial cells. The fluorescent intensity of nitrotyrosine (NT), a specific marker of ONOO~- generation, in BPAECs represented the content of endogenous ONOO~- generation. The fluorescent intensity of NT and the number of NT positive cells were detected with flow cytometry (FCM), and the percentage of NT positive cells was calculated.The results are as follows. (1) LPS (1,5 and 10 μg/ml) caused a marked increase in fluorescent intensity of NT in a dose-dependent manner, which was significantly increased compared to the vehicle group (P0.01). The number and percentage of NT positive cells were markedly increased (both P0.05 vs vehicle group). Aminoguanidine(AG), a selective inhibitor of inducible nitric oxide synthase(iNOS), inhibited LPS-induced increase in fluorescent intensity of NT in BPAECs. However, the number and percentage of NT positive cells had a tendency to reduce. (2) LPS brought about an enhancement in MDA content and the activity of LDH in cultured supernatant. AG reversed the enhancement in MDA content induced by LPS(P0.01). In contrast, AG had a marginal effect on the activity of LDH. (3) LPS induced an increase in apoptotic rate in BPAECs in a dose-dependent manner. The number of apoptotic cells markedly increased as well. Some BPAECs stained with fluorescent probe ethidium bromide showed morphological features of apoptosis with chromatin condensation and nuclear fragmentation. AG reduced the apoptotic rate and the number of apoptotic cells, both of which were still higher than those of vehicle group (P0.05). LPS led to inhibition of mitochondrial respiration and membrane potential in an accumulation manner. In conclusion,LPS caused injury to cultured BPAECs and increased the production of ONOO~-. The cytotoxicity of LPS may be mediated by the endogenous ONOO~-.
【Fund】: This work was supported by the National Natural Science Foundation of China (No. 39870317).
【CateGory Index】: R363
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