特异性抑制转化生长因子β激活激酶1活性对妊娠期糖尿病孕妇外周血单核细胞炎症通路的影响

Influence of specifical inhibition of TAK1 activity in inflammatory pathway of mononuclear cells in peripheral blood of pregnant women with gestational diabetes mellitus

摘要:

目的  探讨特异性抑制转化生长因子β激活激酶1（TAK1）活性的变化对妊娠期糖尿病（GDM）孕妇外周血单核细胞炎症通路的影响。方法  选取GDM孕妇及正常孕妇各30例，采集外周静脉血，分离单核细胞和血浆。依据细胞加药组别的不同，分为LPS刺激组（L组）、脂多糖组（LPS组）、TAK1抑制剂组（LT组）、TAK1抑制剂组（T组）及空白对照组（W组），培养、加药后Western blot检测TAK1及核转录因子-κB（NF-κB）蛋白的表达量，酶联免疫吸附法（ELISA）检测血浆中LPS浓度及各组炎症因子[白介素-1（IL-1）、白介素-10（IL-10）、肿瘤坏死因子-α（TNF-α）]水平，比较各组结果的差异，探讨TAK1的作用。结果  GDM孕妇与正常孕妇比较，血浆LPS浓度升高；TAK1及NF-κB蛋白表达量增加，炎症因子（IL-1、IL-10、TNF-α）水平升高，差异有统计学意义（P <0.05）。各小组间比较，L组相较于其他组的TAK1、NF-κB蛋白表达量增加，炎症因子（IL-1、IL-10、TNF-α）表达水平升高，差异有统计学意义（P <0.05）；正常孕妇中，LT组、T组、W组NF-κB蛋白表达量比较，差异无统计学意义（P >0.05），LT组、T组未检测到TAK1明显表达，可检测到W组少量表达，LT组相对于T组，TNF-α、IL-1、IL-10表达水平升高（P <0.05），LT组、T组与W组上清炎症因子表达水平比较，差异无统计学意义（P >0.05）；GDM孕妇中，LT组、T组、W组TAK1及NF-κB蛋白表达量比较，差异无统计学意义（P >0.05），LT组相对于T组，TNF-α、IL-1、IL-10表达水平升高（P <0.05），Pearson相关性分析发现，TAK1、NF-κB蛋白表达量与炎症因子（IL-1、IL-10、TNF-α）水平呈正相关（P <0.05）。结论  TAK1可能参与GDM孕妇外周血单核细胞炎症通路的形成，抑制TAK1的活性，可以下调通路下游关键介质NF-κB及炎症因子的表达。

Abstract:

Objective To investigate the influence of changes of TGF-β-activated kinase 1 (TAK1) activity in the inflammatory pathway of mononuclear cells in the peripheral blood of pregnant women with gestational diabetes mellitus (GDM). Methods Totally 30 GDM pregnant women and 30 normal pregnant women were selected. Peripheral venous blood was drawn, and mononuclear cells and plasma were separated. They were divided into lipopolysaccharide (LPS)-stimulation group (L group), LPS and TAK1-inhibitor group (LT group), TAK1-inhibitor group (T group) and control group (W group). After culture, Western blot was used to detect the expressions of TAK1 and NF-κB, and ELISA was used to detect the plasma concentration of LPS and the expression levels of inflammatory factors (IL-1, IL-10 and TNF-α). The differences between GDM and normal pregnant women were compared and the effect of TAK1 was explored. Results Compared with the normal pregnant women, the plasma LPS concentration, the expression levels of inflammatory factors (IL-1, IL-10 and TNF-α), TAK1 and NF-κB increased in the GDM pregnant women (P < 0.05). Compared with other groups, the expression levels of TAK1, NF-κB and inflammatory factors (IL-1, IL-10 and TNF-α) significantly increased in the L group (P < 0.05). In the normal pregnant women, the expression levels of NF-κB had no statistical differences (P > 0.05) among the T group, the LT group and the W group; no obvious expression of TAK1 was detected in the LT group or the T group while a small amount of TAK1 expression was detected in the W group; compared with the T group, the expression levels of inflammatory factors (TNF-α, IL-1 and IL-10) significantly increased in the LT group (P < 0.05). In the GDM pregnant women, the LT group, the T group and the W group had no statistical differences in the expression levels of TAK1 or NF-κB (P > 0.05); compared with the T group, the expression levels of inflammatory factors (TNF-α, IL-1 and IL-10) significantly increased in the LT group (P < 0.05). The Pearson correlation analysis showed that the expression levels of TAK1 and NF-κB were positively correlated with inflammatory factors (IL-1, IL-10 and TNF-α) (P < 0.05). Conclusions TAK1 may be involved in the formation of the inflammatory pathway of mononuclear cells in the peripheral blood of GDM pregnant women. Inhibition of TAK1 activity can down-regulate the expressions of downstream key mediators such as NF-κB and inflammatory factors.