摘要
目的:研究脂肪细胞分泌产物脂联素及游离脂肪酸作用于单核细胞的信号转导途径,探讨脂肪细胞与单核细胞间相互作用的分子机制。方法:收集人单核细胞体外培养,加入终浓度分别为0.1、0.5、1.0mmol/L硬脂酸(stearic acid,SA),24h后收集上清液,ELISA法检测上清液肿瘤坏死因子-α(tumor necrosis factor alpha,TNF-α)、白细胞介素-6(interleukin-6,IL-6)浓度;上述浓度SA刺激单核细胞2h后,免疫组化检测单核细胞核转录因子κB(nuclear transcription factor κB,NF-κB)核移位;加入不同浓度脂联素球状结构域(globular adiponectin,gAd)孵育单核细胞6h,SA刺激,2h后检测NF-κB核移位变化,24h后检测上清液TNF-α、IL-6浓度。不同浓度gAd孵育单核细胞24h后用流式细胞术检测Toll样受体4(Toll-like receptors 4,TLR4)数目的变化。结果:SA刺激单核细胞可引起NF-κB核移位及TNF-α、IL-6分泌增加,gAd可阻断SA这一效应。gAd孵育单核细胞可导致TLR4数目显著下调。结论:脂肪细胞分泌SA及脂联素通过TLR4调节单核细胞活性,脂联素通过TLR4阻断SA对单核细胞的激活作用。
AIM： To explore the molecular basis and signaling pathways of the action of adiponectin secreted by adipocytes and free fatty acids on monocytes. METHODS： Human monocytes were collected, cultured and treated with various concentrations （0. 1,0. 5 and 1.0 mmol/L） of stearic acid （SA） in vitro. Nuclear transcription factor κB （NF - κB） of human monocytes were detected by immunohistochemistry 2 h after SA treatments. Tumor necrosis factor alpha （TNF- α） and interleukin- 6 （IL- 6） in supernatants were measured by ELISA and expression of Toll- like receptors 4 （TLR4） on the monocyte membrane was monitored by the flow cytometry at 24 h. The above measurements were repeated after preincubated in various concentrations of globular adiponectin （gad; 10. 0 and 20. 0 mg/L） for 6 h and treated with SA （0. 5 mmol/L） for 2 h or 24 h. RESULTS： SA enhanced the activation of NF - κB and increased the production of TNF - α and IL - 6 in human monoeytes, gAd inhibited the SA effects and down - regulated the expression of TLR4. CONCLUSION ： Adiponectin inhibits the effects of stearic acid on monocytes by down - regulation of TLR4 expression.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2008年第11期2117-2120,共4页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.30872541)
华中科技大学协和医院院内课题资助项目(No.2006-32)
