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目的:探讨中药肺岩宁方含药血清对肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)诱导的人肺癌细胞系A549诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)和环氧合酶2(cyclooxygenase-2,COX-2)表达的影响。方法:制备肺岩宁方含药血清,以肺癌细胞系A549为靶细胞,实时聚合酶链反应法及蛋白质免疫印迹法检测肺岩宁方含药血清对TNF-α诱导的iNOS及COX-2表达的影响;二氨基乙酰乙酸荧光素检测NO产物;人iNOS重组质粒用脂质体转染法导入A549细胞24h后,肺岩宁方含药血清治疗24h,采用双荧光素酶报告基因系统测定肺岩宁方含药血清对TNF-α诱导的细胞核iNOS转录活性的影响。结果:与对照血清比较,TNF-α明显提高iNOS和COX-2的表达,与肺岩宁方含药血清共同作用后,显著抑制二者的表达(P<0.01,P<0.01);肺岩宁方含药血清显著降低了A549细胞经iNOS重组质粒转染后的转录活性(P<0.01,P<0.01)及NO产物的产生。结论:肺岩宁方含药血清抑制肺癌细胞的部分机制可能是抑制iNOS和COX-2基因的活性。
OBJECTIVE: To investigate the effects of the Chinese herbal medicine Feiyanningfang serum on the expression of inducible nitric oxide synthase (iNOS) in human lung cancer cell line A549 induced by tumor necrosis factor-α (TNF-α) and Cyclooxygenase-2 (cyclooxygenase-2, COX-2) expression. Methods: The lung-ning Fang containing serum was prepared. Lung cancer cell line A549 was used as target cell. Real-time polymerase chain reaction and Western blotting were used to detect the effect of Feiyanningfang serum on TNF-α-induced iNOS and COX-2 The expression of NO was detected by fluorescein diaminoacetoacetate. Human iNOS recombinant plasmids were transfected into A549 cells with lipofectamine transfection for 24 h. After 24 h of treatment with Feiyanning containing serum, the dual luciferase reporter gene system was used Effect of Feiyanningfang Serum on Nuclear Transcription of iNOS Induced by. Results: Compared with the control serum, TNF-α significantly increased the expression of iNOS and COX-2, and significantly reduced the expression of TNF-α (P <0.01, P <0.01) Ning Fang containing serum significantly reduced the transcriptional activity of A549 cells (P <0.01, P <0.01) and NO production after transfection with iNOS recombinant plasmid. Conclusion: Part of the mechanism of lung Yanning decoction inhibiting lung cancer cells may be the inhibition of iNOS and COX-2 gene activity.