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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 investigate the effect of Lipoxygenin A4 (LXA4), ProD1 and ResolvinD1 on nuclear factor κB (NFκB) activity and its mechanism. Methods The Chinese hamster ovary cells stably expressing the NFκB luciferase reporter gene were stimulated with LPS, HSP70, HMGB1 or S100A4 by pretreatment with 100 nmol / L LXA4, ProD1 and RvD1 for 30 min, respectively. The effect of small lipid molecules on agonist-activated NFKB activity was assessed by measuring luciferase activity. The content of TNFα in the cell culture supernatant was detected by ELISA, and the amount of NFκB in the nucleus was detected by Western blotting. Results LPS, HSP70, HMGB1 and S100A4 significantly upregulated NFκB activity and increased the amount of TNFα secreted by cells. LXA4, ProD1 and RvD1 significantly inhibited NFκB activation, decreased the level of TNFα secreted by cells, and decreased NFκB entry. Conclusions LXA4, ProD1 and RvD1 significantly inhibit the activation of NFκB by a variety of agonists. The possible mechanism may be related to its ability to reduce the nuclear import of NFκB. These small lipid molecules have the potential for further development and research in the development of novel anti-inflammatory drugs.