摘要
目的:探讨肥胖及伴糖尿病患者血清对人源单核细胞Toll样受体4/转录因子-κB(Toll-like receptor 4/nuclear factor-κB p65,TLR4/NF-κB p65)信号通路的活化作用,了解炎症性免疫反应在肥胖症中的可能作用机制。方法:正常对照组、单纯性肥胖组、肥胖伴糖尿病组每组20例,分离受试者血清,分别用各组血清干预THP-1细胞48 h后,Western印迹和免疫荧光检测THP-1细胞内NF-κB p65蛋白表达;RT-PCR检测TLR4 mRNA的表达;ELISA检测外周血血清干预后细胞培养上清单核细胞趋化因子-1(monocyte chemotactic protein 1,MCP-1)浓度。结果:单纯性肥胖组、肥胖伴糖尿病组血清干预THP-1细胞后,细胞内NF-κB p65磷酸化蛋白、TLR4 mRNA表达水平较正常对照组均有增高,且肥胖伴糖尿病患者高于单纯性肥胖患者(P<0.05),肥胖症患者NF-κB p65的表达存在核转移现象。正常对照组、单纯性肥胖组和肥胖伴糖尿病组血清孵育THP-1细胞48 h后,培养液上清中MCP-1的浓度分别为(26.4±3.9),(45.8±10.0),(58.0±15.3)pg/mL,肥胖症患者患者明显高于正常对照组,且肥胖伴糖尿病患者明显高于单纯肥胖患者(P<0.05)。结论:单纯性肥胖及肥胖的糖尿病患者的血清可不同程度地诱导单核细胞功能紊乱,这种功能紊乱可能与TLR4/NF-κB信号通路的激活有关。
Objective: To investigate the effects of serum from the obesity patients and obesity patients with Diabetic mellitus on toll-like receptor 4/Nuclear factor-κB p65(TLR/NF-κB) pathway in human THP-1 monocytes and to explore the inflammatory immune response in obesity. Methods: Peripheral serum was isolated from healthy volunteers(the control group), the obesity patients(Ob group) and the obesity patients with diabetic mellitus(the Ob with DM group), respectively, 20 in each group. THP-1 monocytes were incubated with the serum for 48 h. The monocytes and culture supernatant were collected. The phosphorylation level of NF-κB p65 protein in THP-1 monocytes was evaluated by Western blot as well as immunofluorescence assay. The TLR4 mRNA expression was evaluated by RT-PCR. ELISA was used to measure the monocyte chemotactic protein-1(MCP-1) levels in the culture supernatant.Results: In the presence of serum, the obesity group and the obesity with diabetic mellitus group showed the up-regulated phosphorylation level of NF-κB p65 protein and TLR4 mRNA expression in THP-1 monocytes compared with the healthy control group(both P<0.05), and the MCP-1 levels in the obesity patients were up-regulated significantly compared with the healthy control group [healthy control group(26.4±3.9) pg/mL, Ob group(45.8±10.0) pg/mL, Ob with DM group(58.0±15.3) pg/mL; P<0.05]. These parameters were further up-regulated in the obesity patients with diabetic mellitus patients.Conclusion: The serum from the obesity patients or the obesity patients with diabetes can induce monocyte dysfunction, which might be related to the activation of TLR4/NF-κB signaling pathway.
出处
《中南大学学报（医学版）》
CAS
CSCD
北大核心
2014年第9期917-923,共7页
Journal of Central South University ：Medical Science
基金
贵州省科技攻关项目[黔科合SY字(2012)3116号]
贵州省科学技术基金项目[黔科合J字LKZ(2013)53号]
遵义医学院博士启动基金(F-588)
重庆市卫生局重点项目(2010-1-16)~~
