TLR4乙酰化对LPS-TLR4-NF-κB通路的影响及其在妊娠期糖尿病发病机制中的作用  

Effect of TLR4 acetylation on LPS-TLR4-NF-κB pathway and its role in pathogenesis of gestional diabetes mellitus

作　　者：李松[1] 丛林[1] 袁静[1] 方慧琴[1] 陈薇[1] 李琴[1] 杨琴[1] 

机构地区：[1]安徽医科大学第一附属医院妇产科产前诊断中心,合肥230022

出　　处：《安徽医科大学学报》2016年第1期26-30,共5页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金项目(编号:1208085MH172);安徽高校省级自然科学研究项目(编号:KJ2011Z215)

摘　　要：目的研究Toll样受体4(TLR4)乙酰化在妊娠期糖尿病(GDM)孕妇外周血单核细胞(PBMC)中的表达及其在内毒素(LPS)-TLR4-核因子κB(NF-κB)通路中的作用,进一步探讨GDM的炎症发病机制。方法选取正常孕妇、GDM孕妇各30例,抽取静脉血15 ml,密度梯度离心法分离PBMC进行体外培养,并保留血清用于检测LPS的量;将研究分成正常组(对照组)、正常+LPS组、GDM组和GDM+LPS组。PBMC用于免疫共沉淀法、Western blot法检测TLR4乙酰化和NF-κB蛋白的表达量;上清液用于ELISA法检测炎症因子肿瘤坏死因子α(TNF-α)、白介素-1(IL-1)、白介素-10(IL-10)的水平。结果正常孕妇、GDM孕妇体内LPS量的差异有统计学意义(P<0.05)。GDM组发生TLR4乙酰化变化,而对照组未检测到TLR4乙酰化;LPS干预后,正常+LPS组、GDM+LPS组TLR4乙酰化的程度明显增加,且GDM+LPS组TLR4乙酰化程度明显高于GDM组和正常+LPS组,差异有统计学意义(P<0.05)。4组NF-κB的表达和炎症因子TNF-α、IL-1、IL-10水平的差异均有统计学意义(P<0.05)。各组TLR4乙酰化与NF-κB蛋白表达量之间具有正相关性(P<0.05)。结论TLR4乙酰化通过影响LPSTLR4-NF-κB通路的活化介导炎症因子的释放,促进孕妇体内抗炎-致炎的失衡,从而参与GDM的发生。Objective To study TLR4 acetylation for analysing the expression in peripheral blood mononuclear cells of gestional diabetes mellitus（GDM） patients, and the impact on LPS-TLR4-NF-KB pathway, discussing the in- flammatory pathogenesis of GDM. Methods 30 normal pregnant women and 30 GDM patients were picked and 15 ml peripheral blood was drawn per subject respectively. Peripheral blood mononuclear cells（PBMC） were extracted via density gradient centrifugation and were cultured in vitro, Serum was used for detecting the level of lipopolysac- charide（LPS）. The research was divided into four groups ： normal group （ control group）, normal ＋ LPS group, GDM group and GDM ＋ LPS group. PBMC were adopted into immunopreeipitation and Western blot was used to de- tect the expression of TLR4 acetylation, the supernatant was used in ELISA for detecting different levels of inflam- matory cytokines, TNF-ct, IL-1 and IL-10. Results Levels of LPS were statistically significant in woman with GDM and normal pregnant woman（P 〈 0.05）. TLR4 acetylation appeared in GDM group, and was negative in the con- trol group. After the LPS intervention, the degree of TLR4 acetylation was enhanced evidently in both normal ＋ LPS group and GDM ＋ LPS group. And the latter was obviously higher than that of GDM group and normal ＋ LPS group （P 〈 0. 05）. The differences among levels of these four inflammatory cytokines,TNF-α,IL-1 and IL-10, had stas- tistical significance（P 〈 0. 05）. There was a positive correlation between TLR4 acetylation and NF-KB protein ex- pression levels （P 〈 0.05 ）. Conclusion TLR4 acetylation mediates the release of inflammatory cytokines by influ- encing the activation of LPS-TLR4-NF-KB pathway, which contributes to promoting antiinflammatory-proinflammato- ry imbalance in pregnant woman, thereby involves in the occurrence of GDM.

关 键 词：妊娠期糖尿病 炎症 TOLL样受体4 乙酰化 

分 类 号：R714.5[医药卫生—妇产科学]