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目的 :探讨炎症时阿司匹林 (AS)对内皮细胞一氧化氮 (NO)的产生及诱导型一氧化氮合酶 (iNOS)基因表达的抑制作用。方法 :Griess法测上清液NO-2 /NO-3 水平、黄递酶法测NOS活性、常规生化法测乳酸脱氢酶(LDH)、丙二醛 (MDA)浓度 ,染料排除法测细胞活力 ,RT -PCR技术分析iNOSmRNA水平。结果 :白介素 (IL) - 1β、肿瘤坏死因子 (TNF) -α、γ -干扰素 (INF)联用脂多糖 (LPS)诱导后上清液中NO-2 /NO-3 由 (4 2 7± 0 75 ) μmol/L增加到(9 35± 1 2 5 ) μmol/L ,对内皮细胞造成明显的损伤。但 3mmol/LAS组NO生成及NOS活性明显降低 ,LDH释放率及MDA浓度下降 ,细胞存活率上升 ,与NO诱导组相比差异显著。并随AS剂量的增加对NO的抑制及对细胞的保护作用更加明显 ,但AS对生理水平的NO没有抑制作用。同时发现 10mmol/L浓度以下AS对iNOSmRNA表达水平没有影响 ;但 10 - 2 0mmol/L的AS则可在转录水平上抑制iNOSmRNA的表达。并观察到水杨酸钠及消炎痛不具有抑制NO产生的作用。结论 :AS具有明显抑制IL - 1β、TNF -α、γ -INF及LPS诱导NO生成的作用 ,从而保护血管内皮细胞避免炎症时高浓度NO的损伤。
Objective: To investigate the inhibitory effect of aspirin on endothelial nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) gene expression in inflammatory conditions. Methods: The level of NO-2 / NO-3 was measured by Griess method, the activity of NOS was detected by enzymolysis method, the concentration of lactate dehydrogenase (LDH) and malondialdehyde (MDA) Vitality, RT-PCR technique to analyze iNOS mRNA levels. Results: NO - 2 / NO - 3 in the supernatant induced by interleukin - 1β, tumor necrosis factor (TNF) - α and interferon - γ (INF) combined with lipopolysaccharide (LPS) ± 0 75) μmol / L to (9 35 ± 125) μmol / L, causing obvious damage to endothelial cells. However, the NO production and NOS activity in 3mmol / LAS group were significantly decreased, the release rate of LDH and the concentration of MDA decreased, and the cell survival rate increased significantly compared with NO induced group. With the increase of AS dose, the inhibition of NO and the protective effect on cells were more obvious, but AS did not inhibit the physiological level of NO. At the same time, it was found that AS concentration below 10 mmol / L had no effect on the expression of iNOS mRNA; however, 10 - 20 mmol / L AS inhibited the expression of iNOS mRNA at the transcriptional level. It was observed that sodium salicylate and indomethacin did not inhibit NO production. CONCLUSION: AS can significantly inhibit NO production induced by IL - 1β, TNF - α, γ - INF and LPS, thereby protecting vascular endothelial cells from high concentration of NO during inflammation.