摘要
目的：研究游离脂肪酸对3T3-L1脂肪细胞核因子NF—κBp65表达及转位的影响，探讨游离脂肪酸诱导胰岛素抵抗的分子机制．方法：诱导成熟的3T3-L1脂肪细胞与0.3，0．5，1．0mmol／L的软脂酸（PA）培养6—24h，用葡萄糖氧化酶法检测培液中的葡萄糖消耗量，以2-脱氧-[^3H]-D-葡萄糖摄入法观察葡萄糖的转运率，用Western blot检测总NF—κBp65蛋白及核NF—κBp65蛋白的表达，用激光扫描共聚焦（CLSM）对NF—κBp65进行定位显示．结果：0．3—1．0mmol／L软脂酸作用6—24h后，3T3-L1脂肪细胞的葡萄糖消耗明显减少（3．03±0．34，2．71±0．36，2．64±0．25mmol／L），呈时间剂量依赖效应，其作用不需要胰岛素的存在：0．3—1．0mmol／L软脂酸作用6—24h显著减少3T3-L1脂肪细胞胰岛素刺激的葡萄糖转运率（64％，33％，32％），呈时间剂量依赖效应；核NF-κBp65蛋白表达明显增加，CLSM显示NF-κBp65核转位增加．但软脂酸对3T3-L1脂肪细胞总NF—κBp65蛋白的表达无明显影响．结论：游离脂肪酸可以诱导胰岛素抵抗，其分子机制可能与FFAs刺激NF—κB的活化转位调节相关基因的表达有关．
AIM： To investigate the effect of free fatty acids （FFAs） on nuclear factor-κB p65 （NF- κBp65） expression and translocation in 3T3-L1 adipocytes and its possible molecular mechanism.
METHODS： 3T3-L1 adipocytes were treated for 6 to 24 hours with palmic acid （0.3, 0.5, 1.0 mmol/L） to induce insulin resistance. Glucose oxidase method was employed to measure the glucose consumption in the medium and 2-deoxy--[^3H]-D-glucose method was used for the determination of glucose uptake. Western blot was used to determine the protein expression of total NF-κBp65 and nuclear NF-κBp65. The distribution of NF-κBp65 was investigated by confocal laser scanning microscopy （CLSM）.
RESULTS： After the treatment with 0.3, 0.5 and 1.0 mmol/L of palmic acid for 6 to 24 hours, the glucose consumption （3.03 ± 0.34, 2.71 ± 0.36, 2.64 ± 0.25 mmol/L） and the insulin-stimulated glucose transport of 3T3-L1 adipose cells （64%, 33%, 32%） were decreased in a dose- and time-dependent manner. Both the expression of nuclear NF-κBp65 and nuclear translocation of NF-κBp65 were increased in a dose- and time-dependent manner. But the total NF-κBp65 protein expression had no marked changes during this study.
CONCLUSION： FFAs can induce insulin resistance, the molecular mechanism of which might be associated with the activation and translocation of NF-κBp65.
出处
《世界华人消化杂志》
CAS
北大核心
2007年第15期1706-1712,共7页
World Chinese Journal of Digestology
基金
国家自然科学基金
No.30371816~~
