IKKε is an IκB kinase participating in the control of NF-κB and type Ⅰ IFN signal pathways in mammals.However,the function of IKKε in regulating immune response is largely unknown in teleost.Herein,an IKKε homol...IKKε is an IκB kinase participating in the control of NF-κB and type Ⅰ IFN signal pathways in mammals.However,the function of IKKε in regulating immune response is largely unknown in teleost.Herein,an IKKε homologue named AjIKKε was characterized in Japanese eel(Anguilla japonica).AjIKKε has an N-terminal kinase domain,a ubiquitin-like domain,and a coiled coil-containing domain(CC),which is conserved and similar to its counterpart in mammals.Expression analysis showed that AjIKKεcould be up-regulated in kidney,spleen,and particularly in liver under the stimulation of poly Ⅰ:C,LPS,and Aeromonas hydrophila infection.In vitro,the mRNA levels of AjIKKεwere significantly provoked in eel liver cells stimulated by LPS and poly I:C,or the different concentrations of A.hydrophila.The overexpression of AjIKKε could not only induce a significantly higher level of promoter activity of human NF-κB,AP-1,and IFN-β in a dose-dependent manner but also up-regulate the activation of promoters of Japanese eel cRel,AP1,IL6,IFN4,IRF3,and IRF7 in HEK293 cells.RNAi studies showed that after AjIKKε was knocked down,the expression levels of IL1,IL6,TNFα,c-Jun,IFN2,IFN3,MX1,MX2,and IRF3 genes were significantly down-regulated in liver,spleen,and kidney of Japanese eels.In addition,the mutants of AjIKKε-K39A,AjIKKε-S174A,and AjIKKε-ΔCC failed to activate Japanese eel IFN4,IRF3 and human IFN-β promoters in HEK293 cells.Collectively,these results suggest that AjIKKεmay function as a positive regulator of NF-κB,MAPK,and IRF3-mediated type Ⅰ IFN signaling pathways related to immune response evoked by bacterial and viral infection.展开更多
目的探讨烙灸治疗早期激素性股骨头坏死(steroid-induced osteonecrosis of the femoral head,SONFH)的作用机制。方法将新西兰兔按随机数表法分为空白组、模型组、烙灸组、抑制剂组,每组6只,采用内毒素联合激素的方法制备SONFH模型。...目的探讨烙灸治疗早期激素性股骨头坏死(steroid-induced osteonecrosis of the femoral head,SONFH)的作用机制。方法将新西兰兔按随机数表法分为空白组、模型组、烙灸组、抑制剂组,每组6只,采用内毒素联合激素的方法制备SONFH模型。烙灸组进行4周的烙灸治疗,抑制剂组给予IκBα磷酸化抑制剂Bay11-7082(1 mg·kg-1)腹腔注射3周。干预前后分别观察实验兔一般状况变化;HE染色观察各组实验兔股骨头组织病理学变化;Western blot和RT-qPCR分别检测股骨头组织中IKKβ、磷酸化IκBα(p-IκBα)蛋白及mRNA的表达变化;ELISA检测血清中核因子κB(nuclear factor kappa-B,NF-κB)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)水平。结果与空白组比较,模型组实验兔逐渐出现精神萎靡、毛发暗淡等情况,股骨头组织骨小梁出现断裂、稀疏等改变,IKKβ、p-IκBα蛋白水平,IKKβmRNA水平及血清中NF-κB、TNF-α、IL-6水平均升高(P均<0.05);与模型组比较,烙灸组股骨头组织病理学改善,IKKβ、p-IκBα蛋白水平,IKKβmRNA水平及血清中NF-κB、TNF-α、IL-6水平均降低(P均<0.05),而IκBα蛋白及mRNA表达均升高(P均<0.05)。结论烙灸可改善早期SONFH骨代谢,缓解骨坏死,其机制与抑制TLR4/NF-κB通路有关。展开更多
文摘IΚB激酶家族(IΚB kinases,IKKs)是NF-ΚB信号途径最为重要的成员之一,已发现其在先天性免疫以及炎症反应的调节中发挥关键作用.除IKKα/β/γ之外,作为IKKs的最新成员,IKKε不仅具有与IKKα/β/γ类似的作用,而且最近的研究数据证实IKKε还能通过凋亡蛋白抑制物(inhibitor of apoptosis proteins,IAPs)调节caspases的活性,参加caspases的凋亡和非凋亡依赖性信号途径.
基金financially supported by“Nature Science Foundation of Fujian Province”(No.2020J01671,2021J01830).
文摘IKKε is an IκB kinase participating in the control of NF-κB and type Ⅰ IFN signal pathways in mammals.However,the function of IKKε in regulating immune response is largely unknown in teleost.Herein,an IKKε homologue named AjIKKε was characterized in Japanese eel(Anguilla japonica).AjIKKε has an N-terminal kinase domain,a ubiquitin-like domain,and a coiled coil-containing domain(CC),which is conserved and similar to its counterpart in mammals.Expression analysis showed that AjIKKεcould be up-regulated in kidney,spleen,and particularly in liver under the stimulation of poly Ⅰ:C,LPS,and Aeromonas hydrophila infection.In vitro,the mRNA levels of AjIKKεwere significantly provoked in eel liver cells stimulated by LPS and poly I:C,or the different concentrations of A.hydrophila.The overexpression of AjIKKε could not only induce a significantly higher level of promoter activity of human NF-κB,AP-1,and IFN-β in a dose-dependent manner but also up-regulate the activation of promoters of Japanese eel cRel,AP1,IL6,IFN4,IRF3,and IRF7 in HEK293 cells.RNAi studies showed that after AjIKKε was knocked down,the expression levels of IL1,IL6,TNFα,c-Jun,IFN2,IFN3,MX1,MX2,and IRF3 genes were significantly down-regulated in liver,spleen,and kidney of Japanese eels.In addition,the mutants of AjIKKε-K39A,AjIKKε-S174A,and AjIKKε-ΔCC failed to activate Japanese eel IFN4,IRF3 and human IFN-β promoters in HEK293 cells.Collectively,these results suggest that AjIKKεmay function as a positive regulator of NF-κB,MAPK,and IRF3-mediated type Ⅰ IFN signaling pathways related to immune response evoked by bacterial and viral infection.
