【摘要】 目的　探讨不同作用机制的抗炎药物对内源性和TNFα诱导的NF -κB的调节作用。方法　利用荧光素酶报告基因测定法检测细胞内NF-κB活化水平,MTT法测定细胞增殖,PI染色流式细胞仪测定细胞凋亡。结果一定浓度的美洛昔康和地塞米松能够明显降低HEK293细胞内源性和10ng·mL-1 TNFα诱导的NF -κB活化, 1×10-9 mol·L-1氢化考的松分别明显增加和降低这两种活化,而吲哚美辛对内源性活化无明显影响。4种药物对HEK293细胞增殖均无明显作用。地塞米松单独或联合TNFα、吲哚美辛联合TNFα能够引起细胞凋亡。结论　不同类型的抗炎药物对内源性和TNFα诱导的NF -κB活化影响不同。提示NF -κB活化调节可能成为药物作用机理研究的有效靶点之一。更多还原

【Abstract】 Aim To investigate the regulatory effects of various anti-inflammatory drugs on both endogenous and TNFα-induced NF-κB activation as well as the relative biological activity. Methods HEK293 cells were cultured in 96-well plate and 6-well plate,treated with meloxicam, indomethacin, dexamethasone and hydrocortisone, without or with 10 ng·mL~-1 TNFα for 24 hours. Then cell proliferation was measured by MTT and cell apoptosis was analyzed by pI stain-flow cytometry. HEK293/κB-luc cells transfected stably with pElam-κB-luc vector, were cultured in 96-well plate and treated as above. Equal amounts of cell lysates were tested for luciferase activity which represents NF-κB activation. Results Endogenous NF-κB activation was present in HEK293 cells and its level can be increased about 2 times by 10 ng·mL~-1 TNFα-induction. Dexamethasone (1×10~-8 mol·L~-1) and meloxicam (1×10~-7-1×10~-6 mol·L~-1) can decrease both endogenous and TNFα-induced NF-κB activation. Hydrocortisone (1×10~-9 mol·L~-1) increases endogenous NF-κB activation but decreases TNFα-induced one significantly. No influence of indomethacin on endogenous NF-κB activation was observed. However, its influence on TNFα-induced NF-κB activation is needed for further study. Cell apoptosis was observed after treatment with TNFα and 1×10~-8, 1×10~-6 mol·L~-1 dexamethasone and 1×10~-7 mol·L~-1 indomethacin, or only with dexamethasone. No significant effect of these anti-inflammatory drugs on cell proliferation was observed. Conclusion Various anti-inflammatory drugs differ in their ability to regulate NF-κB activation in HEK293 cells, which indicates that NF-κB activation might be a potential useful target to study mechanism and for drug screening.更多还原