摘要
目的观察氧化物质第三丁基过氧化氢(t-BHP)对3T3-L1脂肪细胞表达单核细胞趋化蛋白-1(MCP-1)、1型纤溶酶原激活物抑制物(PAI-1)、脂联素的影响初步探讨氧化应激引发脂肪细胞功能障碍的可能作用机制。方法培养3T3-L1脂肪细胞,将其诱导分化为成熟的脂肪细胞,不同浓度t-BHP(100,200,300,400,500μmol/L)作用10min后继续培养。MTT比色法检测24h时3T3-L1脂肪细胞的存活率;荧光定量PCR检测8h时MCP-1、PAI-1、脂联素的表达,Real-time PCR检测t-BHP(500μmol/L,10min)作用于3T3-L1脂肪细胞后不同时间点(2,4,8,12,24h)MCP-1、PAI-1、脂联素的表达。结果 (1)t-BHP对3T3-L1脂肪细胞的存活状态无明显影响;(2)t-BHP可剂量依赖性的降低脂联素而增加MCP-1、PAI-1mRNA的表达;(3)500μmol/L的t-BHP对脂联素、MCP-1、PAI-1的调节具有时间相关性。结论氧化损伤能够促进MCP-1、PAI-1的表达、抑制脂联素的表达,这是其介导脂肪细胞功能障碍的可能机制之一。
Objective To study the effects of the oxidant t-BHP on the expression of adipokines（MCP-1,PAI-1,adiponectin） in 3T3-L1 adipocytes and to discuss the possible mechanisms underlying adipocytes dysfunction induced by oxidative stress.Methods 3T3-L1 pre-adipocytes were differentiated into adipocytes.At 24 h after exposure to t-BHP at concentrations ranging from 100 to 500 mmol/L for 10min,Cell viability was determined by Methylthiazole tetrazolium（MTT）;The mRNA levels of MCP-1,PAI-1and adiponectin were measured by quantitative real-time reverse transcription-polymerase chain reaction（Real Time-PCR） at 8 h after t-BHP treatment,Real Time-PCR was used to detect MCP-1,PAI-1 and adiponectin in cells treated with 500 mmol/L t-BHP for 10 min at different time points（2,4,8,12,24 h）.Results（1）A treatment of t-BHP has little impact on 3T3-L1 adipocytes cell viability.（2）Quantitative Real-Time PCR demonstrated that the mRNA level of adiponectin was down-regulated and the expressions of the MCP-1 and PAI-1 were markedly elevated dose-dependently after the treatment with t-BHP.（3）A 10 min exposure to 500 mmol/L t-BHP affected mRNA expressions of adiponectin,MCP-1 and PAI-1 in 3T3-L1 adipocytes cells in a time dependent manner.Conciusion Oxidative stress increased MCP-1,PAI-1 mRNA expression and decreased gene expression of adiponectin,all of which could result in adipocytes dysfunction.
出处
《福建医科大学学报》
2012年第1期1-6,共6页
Journal of Fujian Medical University
基金
福建省卫生厅青年科研课题(2009/1/17)
