糖尿病患者树突细胞中NF-κB活性改变及检测TNF-α意义  

Dendritic cells in patients with diabetes NF-κB activity and TNF-α significance Detected

作　　者：张洪德[1] 张静春[2] 孙景春[2] 柳忠辉[1] 

机构地区：[1]吉林大学白求恩医学院,吉林长春130021 [2]吉林大学中日联谊医院

出　　处：《中国实验诊断学》2009年第10期1402-1403,共2页Chinese Journal of Laboratory Diagnosis

摘　　要：目的探讨糖尿病患者血清肿瘤坏死因子α(TNF-α)水平及外周血树突细胞(DC)中NF-κB活性变化的意义。方法无偿献血员10例为正常对照组(N组),糖尿病患者20例分为IDM组和ODN组各10名。外周血分离单核细胞(PBMC),加入IL-4、GM-CSF对DC进行诱导培养,同时向ODN组中加入NF-κB特异性低脱氧核苷酸(ODN)培养细胞,采用ELISA法测定血液及培养上清中TNF-α的含量。免疫印迹分析检测DC中NF-κB p65的磷酸化水平,β-actin作为内参。结果糖尿病组患者的血清TNF-α水平显著高于正常对照组(P<0.001);培养物上清液TNF-α水平的测定结果为:N组和ODN组显著低于IDM组(P<0.05);N组和ODN组水平无显著差异(P>0.05);在N组和ODN组DC中NF-κB p65的磷酸化的表达水平显著地低于IDM组(P<0.05);N组和ODN组无显著差异(P>0.05)。结论提示炎症因子TNF-α及NF-κB可能参与糖尿病的病理生理过程。糖尿病患者外周血树突细胞中NF-κB p65的磷酸化水平可以被ODN抑制。Objective diabetes serum tumor necrosis factor α（TNF-α） levels and peripheral blood dendritic cells（DC） in the NF-κB activity significance.Methods 10 cases of unpaid blood donors as normal control group（N group）,20 cases of diabetic patients divided into IDM Group and the ODN group 10.Peripheral blood mononuclear cells（PBMC）,add IL-4,GM-CSF for induction of DC,at the same time add to the ODN group,the low specificity of NF-κB ODN（ODN） cultured cells,using ELISA method blood and culture supernatant of TNF-α content.Western blot analysis detection of DC in the NF-κB p65 phosphorylation levels,βactin as a reference.Results The diabetic group of patients with serum TNFα level was significantly higher than the normal control group（P〈0.001）;culture supernatant levels of TNF-α determination results for：N group and the ODN group was significantly lower than IDM group（P〈0.05）;N group and the level of ODN group no significant difference（P〉0.05）;in the N group and the DC in the ODN group NF-κB p65 phosphorylation expression levels significantly lower than IDM group（P〉0.05）;N ODN group and no significant difference between groups（P〈0.05）.Conclusion The results suggest that inflammatory cytokines TNF-α and NF-κB may be involved in the pathophysiological process of diabetes.Dendritic cells in peripheral blood of diabetic patients in the NF-κB p65 phosphorylation can be inhibited ODN.

关 键 词：糖尿病 核因子ΚB TNF-Α 树突细胞 NF-κB特异性低脱氧核苷酸(ODN) 

分 类 号：R587.1[医药卫生—内分泌]