foxo3a在脓毒血症患者外周血单个核细胞的表达变化及意义  

Changes of forkhead box O3a in the peripheral blood mononuclear of patients with sepsis

作　　者：周超[1] 刘彦[2] 张文锋[3] 陈楠[3] 李培志[3] 

机构地区：[1]成都市第六人民医院肝胆外科,四川成都610051 [2]成都市第五人民医院消化内科,四川成都611130 [3]重庆医科大学附属第二医院肝胆外科,重庆400010

出　　处：《西部医学》2016年第10期1339-1344,共6页Medical Journal of West China

基　　金：国家自然科学基金(81401622)

摘　　要：目的检测叉头蛋白O3a(forkhead box O3a,foxo3a)在脓毒血症患者外周血单个核细胞(peripheral blood mononuclear cell,PBMC)的表达变化,并探讨其在脓毒血症发生过程中的作用。方法分离急性梗阻性化脓性胆管炎(acute obstructive suppurative cholangitis,AOSC)患者(n=27)治愈1周后的PBMC和血清,以及健康志愿者(n=12)的PBMC和血清,分别用蛋白印记法(western blotting,WB)和实时荧光定量聚合酶链反应法(qRT-PCR)检测PBMC中foxo3a、核因子κB p65(NF-κB p65)和核转录因子κB抑制因子(IκB)和蛋白及基因的表达和磷酸化水平;并用酶联免疫吸附法(ELISA)检测血清中脂多糖(LPS)、肿瘤坏死因子α(TNF-α)和白介素10(IL-10)的含量。结果 27例AOSC患者血清中LPS、TNF-α和IL-10的含量明显高于治愈后1周及12例健康志愿者。同时,AOSC患者foxo3a的蛋白和基因的表达量明显低于健康志愿者,但是在治愈1周后,foxo3a的蛋白和基因的含量恢复到正常水平,p-foxo3a与foxo3a的表达呈负相关;另外,NF-κB p65在发病初期增高,治愈1周后基本恢复正常,而IκB及p-IκB表达与foxo3a与p-foxo3a变化相似。这些结果说明在AOSC中foxo3a的磷酸化过程参与调控NF-κB的激活及机体炎症因子的失衡。结论 foxo3a在脓毒血症发生过程中可通过磷酸化降解IκB或直接激活NF-κB,可作为缓解脓毒血症发生初期的治疗靶点之一。Objective To detect the level of forkhead box O3a （foxo3a） in the peripheral blood mononuclear （PBMC） of patients with sepsis before and after treatment and further discuss the function of foxo3a in the process of sepsis. Methods The PBMC was separated from aeute obstructive suppurative eholangitis （AOSC） patients （n=27） on admission （ES）, after cured for 1 week （CG） and healthy volunteers （n= 12） （HV）. The serum levels of lipopolysaecharide （LPS）, tumor necrosis factor α（TNF-a） and interleukin 10 （IL-10） were detected by enzyme linked immunosorbent assay （ELISA） methods. The mRNA levels of foxo3a, nuclear fator κB P65 （NF-κB） and inhibitor of NF-κB （IκB） were detected by real-time quantitative polymerase chain reaction （qRT-PCR）. The protein levels of foxo3a, p-foxo3a, IκB, p-IκB and NF-κB were tested by western blotting （WB）. Results The levels of LPS, TNF-α and IL-10 in ES were significantly higher than that in HV and CG. The mRNA levels of foxo3a and IκB in ES were lower than that in HV and CG, while NF-κB P65 in ES was higher than that in HV and CG. The protein levels of foxo3a and IκB in ES were lower than HV and CG, while p-foxo3a, p-IκB and NF-κB P65 were higher than HV and CG. Conclusion The foxo3a for phosphorylation activation of IκB or directly activation of NF-κB participate the originating process of sepsis and hinting the therapeutic potentialities in the early stage of sepsis.

关 键 词：脓毒血症 叉头蛋白O3a 核转录因子κB抑制因子 

分 类 号：R631.4[医药卫生—外科学]