CAJ | 학술논문

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24 h后,肺岩宁方含药血清治疗24 h,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.
목적:탐토중약폐암저방함약혈청대종류배사인자α(tumor necrosis factor-α,TNF-α)유도적인폐암세포계A549유도형일양화담합매(inducible nitric oxide synthase,iNOS)화배양합매2(cyclooxygenase-2,COX-2)표체적영향.방법:제비폐암저방함약혈청,이폐암세포계A549위파세포,실시취합매련반응법급단백질면역인적법검측폐암저방함약혈청대TNF-α유도적iNOS급COX-2표체적영향；이안기을선을산형광소검측NO산물；인iNOS중조질립용지질체전염법도입A549세포24 h후,폐암저방함약혈청치료24 h,채용쌍형광소매보고기인계통측정폐암저방함약혈청대TNF-α유도적세포핵iNOS전록활성적영향.결과:여대조혈청비교,TNF-α명현제고iNOS화COX-2적표체,여폐암저방함약혈청공동작용후,현저억제이자적표체(P＜0.01,P＜0.01)；폐암저방함약혈청현저강저료A549세포경iNOS중조질립전염후적전록활성(P＜0.01,P＜0.01)급NO산물적산생.결론:폐암저방함약혈청억제폐암세포적부분궤제가능시억제iNOS화COX-2기인적활성.
OBJECTIVE:To study the effect of Feiyanning Decoction (FYN),a compound traditional Chinese medicine,on expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) activated by tumor necrosis factor-α (TNF-α) in human lung adenocarcinoma epithelial cell line (A549).METHODS:A549 cells were incubated with rat serum containing FYN for 24 h.Gene expressions of iNOS and COX-2 were determined by quantitative real-time polymerase chain reaction and Western blot.The iNOS-dependent luciferase reporter was transfected for 24 h and the cells were treated with the reagents for 24 h,then the transcriptional activity of iNOS promoter was detected by luciferase assay.The production of NO was determined by diaminofluorescein-2.RESULTS:FYN significantly inhibited TNF-α-induced expression of iNOS and COX-2 compared with the control group in A549 cells (P＜0.01,P＜0.01).Also,FYN inhibited the transcriptional activity of the iNOS promoter and reduced NO production compared with the control group (P＜0.01,P＜0.01).CONCLUSION:These results suggest that FYN inhibits iNOS and COX-2 activation induced by TNF-α,therefore,it is expected to develop a new strategy to treat lung cancer.