CAJ | 학술논문

目的研究核因子κB抑制物激酶α(IKKα)在肾脏缺血再灌注(IR)诱导的炎性反应中的作用及相关机制.方法用6～8周龄健康雄性C57BL/6小鼠构建肾脏IR模型,采用常规生化法检测Scr、BUN水平,HE染色法观察肾组织学改变,免疫组织化学法、Western印迹法检测肾组织IKKα、p52、RelB、IL-10、IL-18的表达.在肾实质内转入以慢病毒为载体的短发夹RNA (shRNA),抑制IKKα的表达,检测抑制后的肾功能与肾组织形态学改变以及IKKα、p52、RelB、IL-10、IL-18的表达变化.结果 IR损伤组1d、3d、7d血清Scr[(29.80±2.10) μmol/L、(27.00±3.40)μmol/L、(23.00±3.70) μmol/L]和BUN[(9.47±3.50) mmol/L、(11.68±4.30) mmol/L、(13.12±2.10) mmol/L]高于假手术组Scr[(7.30±0.13)μmol/L]和BUN[(8.39±0.30) mmol/L].HE染色显示,IR损伤组存在肾组织病理损害.免疫组化结果显示,Sham组小鼠肾组织抗炎因子IL-10、促炎因子IL-18均呈弱阳性表达,IR损伤后IL-10、IL-18均呈高表达,且IL-10表达呈时间依赖性上调,IL-18表达呈时间依赖性下调.IR损伤后7d内,IKKα、p52、RelB表达均上调,以第3天时最为明显.与IR损伤组相比,慢病毒载体shRNA组肾组织IR损伤第3天的IKKα、p52、RelB表达下调,IL-18表达上调,IL-10表达下调.结论肾脏IR损伤炎性反应消退过程中,NF-κB信号通路被激活,IKKα表达上调.抑制IKKα可能通过下调NF-κB旁路途径家族成员p52、RelB的活性,阻碍肾脏IR损伤炎性反应的消退.
목적 연구핵인자κB억제물격매α(IKKα)재신장결혈재관주(IR)유도적염성반응중적작용급상관궤제.방법 용6～8주령건강웅성C57BL/6소서구건신장IR모형,채용상규생화법검측Scr、BUN수평,HE염색법관찰신조직학개변,면역조직화학법、Western인적법검측신조직IKKα、p52、RelB、IL-10、IL-18적표체.재신실질내전입이만병독위재체적단발협RNA (shRNA),억제IKKα적표체,검측억제후적신공능여신조직형태학개변이급IKKα、p52、RelB、IL-10、IL-18적표체변화.결과 IR손상조1d、3d、7d혈청Scr[(29.80±2.10) μmol/L、(27.00±3.40)μmol/L、(23.00±3.70) μmol/L]화BUN[(9.47±3.50) mmol/L、(11.68±4.30) mmol/L、(13.12±2.10) mmol/L]고우가수술조Scr[(7.30±0.13)μmol/L]화BUN[(8.39±0.30) mmol/L].HE염색현시,IR손상조존재신조직병리손해.면역조화결과현시,Sham조소서신조직항염인자IL-10、촉염인자IL-18균정약양성표체,IR손상후IL-10、IL-18균정고표체,차IL-10표체정시간의뢰성상조,IL-18표체정시간의뢰성하조.IR손상후7d내,IKKα、p52、RelB표체균상조,이제3천시최위명현.여IR손상조상비,만병독재체shRNA조신조직IR손상제3천적IKKα、p52、RelB표체하조,IL-18표체상조,IL-10표체하조.결론 신장IR손상염성반응소퇴과정중,NF-κB신호통로피격활,IKKα표체상조.억제IKKα가능통과하조NF-κB방로도경가족성원p52、RelB적활성,조애신장IR손상염성반응적소퇴.
Objective To reveal the role of inhibitor of nuclear factor kappa B kinase alpha (IKKα) in renal inflammation after renal ischemia-reperfusion (IR) injury and its potential associated mechanism.Methods Ischemia-reperfusion injury models were induced in a total of 24 healthy C57BL/6 male mice.Renal function and histological changes were estimated.The expression and site of IKKα,p52,RelB,IL-10 and IL-18 were determined by immunohistochemistry and Western blotting.After the short hairpin RNA(shRNA)targeting IKKα was injected into renal parenchyma,renal function and protein expressions of IKKα,p52,RelB,IL-10,IL-18 were detected.Results Compared with sham-operated group[Scr(7.30±0.13) μmol/L,BUN (8.39± 0.30) mmol/L],levels of Scr [(29.80± 2.10)μmol/L,(27.00±3.40) μmol/L,(23.00±3.70) μmol/L] and BUN [(9.47±3.50) mmol/L,(11.68 ±4.30)mmol/L,(13.12±2.10) mmol/L] were higher on day 1,3,7 and the injury of kidney was serious in IR injury group.Immunohistochemical expression of both IL-18 and IL-10 were increased.Markedly increased IKKα,p52 and RelB protein expression were noted in experiments from day 1 to day 7 during kidney recovery period,with a peak on day 3 and then decreasing toward baseline after day 7.Compared with IR injury group,low-expression of IKKα by injection of shRNA up-regulated the expression of IL-18 and down-regulated the expression of IKKα,p52,RelB and IL-10.Conclusions The NF-κB pathway is activated and IKKα expression is up-regulated during the kidney ischemiareperfusion injury,low-expression of IKKα may block inflammation resolution via down-regulation of alternative NF-κB pathway family members of both p52 and RelB.