TLR4/NF-κB信号通路介导肥胖症患者血清的致炎作用  

TLR4/NF-κB Signaling Pathway Mediates the Proinflammatory Effect of the Obesity Serum

作　　者：姚岚[1,2] 肖扬[1,2] 刘石平[1,2] 徐爱民[3] 周智广[1,2] 

机构地区：[1]中南大学湘雅二医院代谢内分泌研究所 [2]中南大学糖尿病中心糖尿病免疫学教育部重点实验室,湖南省长沙市410011 [3]香港大学李嘉诚医学院内科系和药理系,香港999077

出　　处：《中国动脉硬化杂志》2010年第11期873-877,共5页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学基金委员会与香港研究资助局联合基金(30831160518);中南大学代谢综合征研究中心项目;湖南省高校科技创新团队资金资助

摘　　要：目的探讨肥胖症患者血清对THP-1单核细胞分泌炎症因子的作用及探讨TLR4/NF-κB信号通路在肥胖症患者血清促炎症作用中的贡献。方法分别用15例单纯性肥胖及伴不同代谢紊乱(血脂紊乱、高血压、高血糖)患者血清孵育THP-1单核细胞株48 h后以及用TLR4单克隆抗体阻断TLR4/NF-κB信号通路后,测定THP-1细胞表面TLR4和细胞内NF-κB p65磷酸化的表达水平及其分泌白细胞介素1β和肿瘤坏死因子α的能力。免疫印迹法检测TLR4蛋白和细胞内NF-κB p65磷酸化蛋白水平;用RT-Q-PCR检测TLR4 mRNA的表达水平;用酶联免疫吸附法检测细胞上清液中白细胞介素1β和肿瘤坏死因子α的水平。结果与正常对照组比较,单纯性肥胖和肥胖症伴代谢紊乱患者血清干预后,THP-1细胞TLR4、NF-κB p65磷酸化蛋白及TLR4 mRNA表达水平以及分泌白细胞介素1β和肿瘤坏死因子α的能力显著升高(P<0.05);THP-1细胞TLR4、NF-κB p65磷酸化蛋白及TLR4mRNA表达水平以及分泌白细胞介素1β和肿瘤坏死因子α的能力随肥胖症代谢紊乱组分的增加而增高,且当TLR4/NF-κB信号通路被阻断后,分泌白细胞介素1β和肿瘤坏死因子α的能力降低。结论单纯性肥胖及肥胖症伴不同代谢紊乱患者的血清可不同程度地诱导THP-1细胞TLR4/NF-κB信号通路的活化,TLR4/NF-κB信号通路在肥胖症患者血清促THP-1分泌炎症因子的作用中起重要作用。Aim To investigate the effect of serum of obese people on the formation process of inflammatory factor secreted by THP-1 cell line,meanwhile,to inquire the role of TLR4/NF-κB signaling pathway on proinflammatory effect of the serum of obesity. Methods THP-1 cell line was cultured from the serum of volunteers with varying degrees of metabolic disorders（4 groups in total,normal people,simple obesity,obesity with hyperglycemia,obesity with hyperglycemia combining hypertension and hyperlipidemia,15 cases of each group） for 48 h and then TLR4/NF-κB signaling pathway is blocked by TLR4 monoclonal antibody.We observed the TLR4 mRNA and protein content on cell surface,the level of NF-κB p65 phosphorylation in THP-1 cell line and IL-1β and TNF-α expression in culture supernatant.TLR4 protein content and the level of NF-κB p65 phosphorylation were measured by Western blotting,TLR4 mRNA was measured by RT-Q-PCR,the expression of IL-1β and TNF-α were detected by ELISA. Results Compared with normal group,The TLR4 mRNA and protein content,the level of NF-κB p65 phosphorylation in THP-1 cell line and IL-1 and TNF-α expression in culture supernatant cultured by the serum from the simple obesity,obesity with metabolic disorders increase,in which the TLR4 mRNA and protein content,the level of NF-κB p65 phosphorylation,the IL-1 and TNF-α expression increased with component of metabolic disorders.While TLR4 was blocked by TLR4 monoclonal antibody,the ability of secreting IL-1 and TNF-α reduced. Conclusion The serum of simple obesity and obesity with varying degrees of metabolic disorders can induce the activation of TLR4/NF-κB signaling pathway on THP-1 cell line.TLR4/NF-κB signaling pathway plays an important part in proinflammatory effect of the serum of obese people.

关 键 词：TOLL样受体4 肥胖症 炎症 代谢紊乱 

分 类 号：R5[医药卫生—内科学]