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| 肿瘤坏死因子-α诱导脂肪细胞胰岛素抵抗模型的建立及评价指标 |
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刘璐1,2, 曹世杰2, 程丽娜1,2, 邱峰2,3, 康宁1,2
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1.天津中医药大学中西医结合学院生物化学教研室, 天津 300193;2.天津市现代中药重点实验室, 天津 300193;3.天津中医药大学中药学院中药化学教研室, 天津 300193
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| 摘要: |
| [目的]确定运用肿瘤坏死因子-α（TNF-α）建立胰岛素抵抗模型的最适时间与剂量，并通过检测TNF-α对葡萄糖转运体4（GLUT4）表达及膜转位的影响探讨模型成立的评价指标。[方法]用不同浓度（5、10、15、20、25 ng/mL）的TNF-α作用于3T3-L1脂肪细胞不同的时间（48、72、96 h），建立胰岛素抵抗模型。另外，分别采用蛋白免疫印迹法（Western blot）和高内涵筛选技术检测模型组细胞GLUT4蛋白表达及分布情况。[结果]与对照组相比，10 ng/mL TNF-α作用3T3-L1脂肪细胞96 h后的模型组的葡萄糖摄取及GLUT4的表达均有显著的降低；加入胰岛素后，对照组的葡萄糖摄取及GLUT4的表达有明显的升高，而模型组无明显变化；胰岛素抵抗模型中，GLUT4膜转位显著降低，差异有统计学意义。[结论]10 ng/mL的TNF-α作用3T3-L1脂肪细胞96 h有利于建立胰岛素抵抗模型。此外，葡萄糖摄取结合GLUT4的蛋白表达及膜转位可作为胰岛素抵抗模型建立的评价标准。 |
| 关键词: 肿瘤坏死因子-α 脂肪细胞 胰岛素抵抗 葡萄糖转运体4 高内涵筛选技术 |
| DOI：10.11656/j.issn.1672-1519.2018.11.14 |
| 分类号:R587.1 |
| 基金项目:国家自然科学基金项目（81430095）。 |
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| Establishment and evaluation of tumor necrosis factor-α induced insulin resistance in 3T3-L1 adipocytes |
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LIU Lu1,2, CAO Shijie2, CHENG Lina1,2, QIU Feng2,3, KANG Ning1,2
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1.Department of Biochemistry, College of Integrated Traditional Chinese and Western Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China;2.Tianjin Key Laboratory of Modern Traditional Chinese Medicine, Tianjin 300193, China;3.Department of Chinese Medicinal Chemistry, College of Traditional Chinese Materia Medica, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China
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| Abstract: |
| [Objective] To determine the optimal time and dose of tumor necrosis factor-α (TNF-α)-induced insulin resistance model and to investigate the effect of TNF-α on the translocation of glucose transporter 4 (GLUT4) influences to explore the evaluation of the establishment of the model.[Methods] The insulin resistance model was established by using TNF-α at different concentrations (5, 10, 15, 20, 25 ng/mL) for different time (48, 72, 96 h) in 3T3-L1 adipocytes. In addition, the expression and distribution of GLUT4 protein were detected by Western blot and high content screening (HCS).[Results] The results showed that the insulin resistance model 3T3-L1 cells were established by treating with 10 ng/mL TNF-α for 96 h. Compared with the control group, the glucose uptake and GLUT4 expression were significantly decreased in the model group. After adding insulin, glucose uptake and GLUT4 expression of control group were significantly increased. Furthermore, the translocation of GLUT4 to the plasma membrane was decreased in the insulin resistance model, differences were statistically significant.[Conclusion] TNF-α can significantly inhibits the uptake of glucose and the translocation of GLUT4 to the plasma membrane in insulin resistance model induced by treating with 10 ng/mL TNF-α for 96 h in 3T3-L1 adipocytes, and therefore supplemented the insulin resistance model established by the evaluation criteria. |
| Key words: tumor necrosis factor-α adipocytes insulin resistance glucose transporter 4 high content screening |
