摘要
目的探讨脂质小分子脂氧素A4(LXA4)、保护素D1(ProD1)、ResolvinD1(RvD1)对核因子κB(NFκB)活性的影响及作用机制。方法稳定表达NFκB荧光素酶报告基因的中国仓鼠卵巢细胞分别由100 nmol/L LXA4、ProD1、RvD1预处理30 min后,细胞被激动剂LPS、HSP70、HMGB1或S100A4刺激。通过检测荧光素酶活性以评价脂质小分子对激动剂激活NFκB活性的作用。细胞培养上清中TNFα的含量由ELISA检测,胞核中NFκB的含量由Western印迹检测。结果 LPS、HSP70、HMGB1和S100A4显著上调NFκB的活性,增加细胞分泌的TNFα的量。LXA4、ProD1、RvD1显著抑制NFκB激活,降低细胞分泌的TNFα含量,减少NFκB的入核。结论 LXA4、ProD1、RvD1显著抑制多种激动剂活化NFκB,其作用机制可能与其能降低NFκB的入核有关,这几个脂质小分子在研制新型抗炎药物方面具有进一步开发和研究的潜力。
Objective To examine effect of lipoxin A4 （LXA4） , protectin D1 （ ProD1 ） or resolvin D1 （ RvD1 ） on the activity of NFKB and their action mechanism. Methods The CHO cells, stably expressing NFKB luciferase reporter gene, were treated with LPS, HSP70, HMGB1 or S100A4, in the presence or absence of 100 nmol/L of LXA4, ProD1 or RvD1 for 30 minutes. The activity of NFKB was detected with the luciferase assay. The content of tumor necrosis factor a （TNFa） in supernatant was measured by enzymelinked immunosorbent assay （ELISA） and the expression of NFkB in the nucleus was detected by immune blotting. Results The activity of NFKB and the level of TNFa in supernatant were significantly upregulated after treatment of the cells with LPS, HSP70, HMGB1 or S100A4, respectively. However, the NFKB activity and concentration of TNFa were lowered in the ceils preincubated with LXA4, ProD1 or RvD1 as compared to the stimulated ceils. Moreover, the lipids significantly decreased the content of NFKB in the nucleus. Conclusion LXA4, ProD1 or RvD1 could significantly inhibit the ligand-stimulated NFKB activity through interfering with NFKB transloeation from cytoplast to nucleus. LXA4, ProD1 and RvD1 showed the potential in the development of new anti - inflammatory therapeutics, which was required further research.
出处
《药学实践杂志》
CAS
2012年第3期185-188,共4页
Journal of Pharmaceutical Practice
基金
国家自然科学基金重点项目(81030056)
国家自然科学基金面上项目(30672468)
中央级公益性科研院所基本科研业务费(2012CHX07)
