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目的 应用硝普钠 (SNP)作为一氧化氮 (NO)供体观察NO对60 Coγ射线照射的人胚肺成纤维细胞 (HELF)增殖活力及其分泌功能的影响 ,以探讨NO的抗纤维化机理。方法 应用MTT比色法、Griess一氧化氮浓度测定法、免疫组化和原位杂交等方法检测成纤维细胞增殖活性及内皮素 (ET)、血管紧张素II(AII)和转化生长因子 β(TGFβ)的表达情况。 结果 低剂量的60 Coγ射线照射对人胚肺成纤维细胞有促增殖作用 ,以 5Gy剂量的促增殖作用最强 ;30h以内 ,硝普钠基本以线性方式释放NO ,平均增幅为 4 2 5μmol L ,初步认为 4～ 5μmol L的NO为抑制成纤维细胞增生的最佳浓度 ;未照射成纤维细胞不表达或仅微弱表达ET、AII和TGFβ ;照后细胞ET和TGFβmRNA及其蛋白表达显著增强 ,AII合成增加。结论 NO供体硝普钠可明显抑制成纤维细胞的过度增殖和分泌活动 ,随浓度的加大 ,抑制作用增强 ,呈明显的剂量依赖关系
Objective To observe the effect of nitric oxide (NO) donor on the proliferation of human embryonic lung fibroblasts (HELF) stimulated by 60Coγ ray and its secretion function by using sodium nitroprusside (SNP) as a nitric oxide (NO) mechanism. Methods MTT assay, Griess NO concentration assay, immunohistochemistry and in situ hybridization were used to detect the proliferation of fibroblasts and the effects of endothelin (ET), angiotensin II (AII) and transforming growth factor β TGFβ) expression. Results Low dose of 60 Co γ -ray irradiation could promote the proliferation of human embryo lung fibroblasts, and promoted the proliferation at the dose of 5 Gy. Within 30 hours, sodium nitroprusside released NO almost linearly with an average increase of 425 micromol L , Initially considered 4 ~ 5μmol L of NO as the best concentration to inhibit fibroblast proliferation; non-irradiated fibroblasts do not express or only weakly expressed ET, AII and TGFβ; according to the cell after ET and TGFβ mRNA and protein expression was significantly enhanced, AII synthesis increased. CONCLUSION NO donor sodium nitroprusside can significantly inhibit the excessive proliferation and secretion of fibroblasts. With the increase of concentration, the inhibitory effect is enhanced in a dose-dependent manner