[目的]本文旨在探讨艾纳香油(BBO)通过核转录因子κB(NF-κB)非经典通路影响花生四烯酸(AA)代谢通路发挥抗炎作用.[方法]采用豚鼠离体回肠法检测BBO对慢反应物质(SRS-A)生成的影响,ELISA法检测BBO对脂多糖(LPS)诱导巨噬细胞前列腺素E2(PGE2)及白三烯 B4(LTB4)产生的影响,qRT-PCR检测BBO对LPS诱导巨噬细胞COX-2、5-LOX、FLAP和RelB表达的影响,Western blot检测BBO对NF-κB非经典通路蛋白肿瘤坏死因子受体作用因子3(TRAF3)、肿瘤坏死因子受体作用因子2(TRAF2)、NF-κB 诱导激酶(NIK)、p100和RelB浓度的影响.[结果]在1mg·mL-1的BBO药物浓度下减弱了豚鼠回肠的收缩张力(P＜0.001),对SRS-A的生成抑制率达到65.34%;与LPS模型组相比,BBO在40-80 μg·mL-1浓度下降低AA代谢通路中PGE2(P<0.05)和LTB4(P<0.05)的浓度,降低COX-2(P<0.05)、5-LOX(P<0.05)和FLAP(P<0.05)的表达;此外,40-80 μg·mL-1浓度下BBO还降低LPS导致的NF-κB非经典通路中TRAF3(P<0.05)、TRAF2(P<0.05)和NIK(P<0.05)的浓度,进一步抑制p100蛋白的磷酸化(P<0.05),同时抑制转录因子RelB的表达(P<0.05)和RelB蛋白的水平(P<0.05),而BBO自身不引起这些基因与蛋白的变化.[结论]BBO可能通过抑制NF-κB非经典信号中调节蛋白TRAF3和TRAF2,转录因子RelB的浓度,造成NIK蛋白的诱导激酶作用受到抑制,进一步抑制了p100蛋白的磷酸化及其与转录因子RelB的结合,从而影响到下游AA通路中重要限速酶COX-2、5-LOX和 FLAP的表达与炎症介质PGE2和LTB4的水平发挥抗炎作用.
[Objective]To study the anti-inflammatory effects of Blumea balsamifera(L.)DC oil(BBO)based on nu-clear factor kappa-B(NF-κB)nonclassical and arachidonic acid(AA)pathway.[Methods]Effects of BBO on the produc-tion of slow reacting substance of anaphylaxis(SRS-A)were detected by the ileal smooth muscle method.The contents of prostaglandin E2(PGE2)and leukotriene B4(LTB4)in lipopolysaccharides(LPS)-induced macrophages were detected by ELISA kit.The expression of COX-2,5-LOX,FLAP and RelB were detected by qRT-PCR.Western blot was performed to detect the effects of BBO on the level of NF-κB nonclassical pathway proteins TNF receptor associated factor 3(TRAF3),TNF receptor associated factor 2(TRAF2),NF-κB-inducing kinase(NIK),p100 and RelB.[Results]The contractile tension of guinea pig ileum was reduced(P<0.001),and the SRS-A production inhibition rate reached 65.34%at 1mg·mL-1 BBO concentration.Compared with LPS group,BBO reduced the concentrations of PGE2(P<0.05)and LTB4(P<0.05),and decreased the expressions of COX-2(P<0.05),5-LOX(P<0.05)and FLAP(P<0.05)in AA pathway at con-centrations of 40-80 μg·mL-1.Moreover,40-80 μg·mL-1 BBO decreased the concentrations of TRAF3(P<0.05),TRAF2(P<0.05),and NIK(P<0.05),and further inhibited the phosphorylation of p100(P<0.05),as well as the level of the transcription factor RelB in genes(P<0.05)and proteins(P<0.05)in nonclassical NF-κB pathway,whereas BBO did not cause such changes.[Conclusion]BBO may potentially exert its anti-inflammatory effects by suppressing the regu-latory proteins TRAF3 and TRAF2 and the transcription factor RelB in NF-κB nonclassical pathway.The inhibitory action extending to the induction kinase function of NIK,further hindering the phosphorylation of p100 and its binding with the transcription factor RelB.Consequently,downstream elements in the AA pathway,including the pivotal rate-limiting en-zymes COX-2,5-LOX and FLAP,were altered.This modulation influences the levels of inflammatory mediators such as PGE2 and LTB4.
何贤芳;王万林;王鸿颖;刘瑞秀;易琼;王鲁
贵州大学药学院,贵州 贵阳 550025||西南特色药用生物资源开发利用教育部工程研究中心,贵州 贵阳 550025贵州省三穗县人民医院,贵州 凯里 556000西南特色药用生物资源开发利用教育部工程研究中心,贵州 贵阳 550025
基础医学
艾纳香油;脂多糖;NF-κB非经典通路;花生四烯酸代谢;抗炎作用
Blumea balsamifera(L.)DC oil;lipopolysaccharides;nonclassical NF-κB pathway;arachidonic acid metabolism;anti-inflammatory effect
《中山大学学报（医学科学版）》 2024 (002)
216-225 / 10
国家自然科学基金(32160849,31760747)
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