摘要
目的:利用NFκ-B转录活性萤光素酶报告系统,建立新的白细胞介素1(interleuk in-1,IL-1)和白细胞介素1受体拮抗剂(interleuk in-1 receptor antagon ist,IL-1 ra)生物学活性的检测方法。方法:运用萤光素酶转录报告系统,选用小鼠胸腺瘤细胞系EL4细胞(EL4的某些亚克隆细胞表面具有高密度IL-1受体表达),以pNFκ-B-luc质粒和内对照pRL-TK质粒共转染,并加入IL-1β激活,以双萤光素酶报告基因检测系统检测IL-1β及其抑制剂IL-1 ra的生物学活性。结果:这种方法检测到IL-1β激活NFκ-B报告基因表达萤光素酶,并且这种激活可被IL-1 ra所抑制,实验重复性好。IL-1β在5μg/L为激活NFκ-B报告基因表达萤光素酶的最佳浓度,并可被IL-1 ra 50μg/L最大抑制。结论:这种检测方法的建立为今后对IL-1β和IL-1 ra的生物学活性检测提供了新的途径。
Objective： To develop a reporter gene system based on transient transfections with a NF-κB responsive reporter gene to detect the bioactivity of IL-1β and IL-1 receptor antagonist. Methods： NF-KB reporter and Dual-Luciferase assays were applied to measure the bioactivity of IL-1β and IL-1 receptor antagonist in mouse EIA cells （ some subclones of EIA cells expressed high level of IL-1 receptor on cell surface）, pNF-κB-luc and pRL-TK, used as an internal control, were co-transfected into EIA cells and then the IL-1β was added. Results： The results indicated that IL-1β was able to induce the expression of this luciferase, which could be blocked by IL-1 receptor antagonist. The optimal dose of IL-1β was 5 μg/L in Dual-Luciferase assay, whose bioactivity can be effectively inhibited by IL-1ra at 50μg/L. Conclusion： We have established a new method to detect the bioactivity of IL-1β and IL-1 receptor antagonist, which can give repeatable results.
出处
《北京大学学报（医学版）》
CAS
CSCD
北大核心
2006年第6期653-656,共4页
Journal of Peking University:Health Sciences
