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【目的】观察TNF-α对3T3-L1成熟脂肪细胞中NYGGF4小鼠同源基因表达水平以及胰岛素刺激下的葡萄糖摄取率的影响。【方法】体外培养3T3-L1前体脂肪细胞,诱导分化为成熟脂肪细胞后,应用不同浓度(10、25、50 ng/mL)重组TNF-α干预成熟脂肪细胞16 h,或以10 ng/mL重组TNF-α分别干预24、487、2 h,采用实时荧光定量RT-PCR技术检测TNF-α干预后3T3-L1脂肪细胞中NYGGF4小鼠同源基因mRNA表达水平;另外,采用[3H]-2-脱氧葡萄糖掺入法检测TNF-α干预24、48、72h后成熟脂肪细胞葡萄糖摄取率。【结果】1)不同浓度TNF-α均能显著上调3T3-L1成熟脂肪细胞中NYGGF4小鼠同源基因表达(P均<0.001),在0~25 ng/mL浓度范围内呈现浓度依赖性,随TNF-α干预浓度增高,NYGGF4小鼠同源基因的表达水平逐渐升高;2)TNF-α对3T3-L1成熟脂肪细胞中NYGGF4小鼠同源基因的表达调节具时间反应性,呈现随干预时间延长该基因表达逐渐上调的特征,10 ng/mL TNF-α干预人成熟脂肪细胞24 hNYGGF4小鼠同源基因mRNA表达水平即显著上调(P<0.001);3)TNF-α干预48 h,3T3-L1成熟脂肪细胞胰岛素刺激的葡萄糖摄取率与未干预组比较下降50%以上,干预72 h,胰岛素刺激的葡萄糖摄取受到进一步抑制。【结论】TNF-α可以上调3T3-L1成熟脂肪细胞中NYGGF4小鼠同源基因的表达,抑制3T3-L1成熟脂肪细胞胰岛素刺激下的葡萄糖摄取。
【Objective】 To observe the effect of TNF-α on the homologous gene expression in NYGGF4 mice and the glucose uptake rate under insulin stimulation in 3T3-L1 mature adipocytes. 【Method】 After the 3T3-L1 precursor adipocytes were cultured in vitro and differentiated into mature adipocytes, the mature adipocytes were treated with recombinant TNF-α at different concentrations (10, 25 and 50 ng / mL) for 16 h or 10 ng / mL recombinant TNF-α were respectively measured for 24,487,2 h. The mRNA expression of NYGGF4 in 3T3-L1 adipocytes was detected by real-time fluorescence quantitative RT-PCR. In addition, 2-deoxyglucose incorporation method was used to detect the glucose uptake rate of mature adipocytes after 24 h, 48 h and 72 h of TNF-α intervention. 【Results】 1) TNF-α at different concentrations could significantly up-regulate the homologous gene expression of NYGGF4 mice in 3T3-L1 mature adipocytes (all P <0.001) in a concentration-dependent manner in the range of 0-25 ng / mL, The expression of homologous genes in NYGGF4 mice increased gradually with the intervention of TNF-α. 2) The expression of NYGGF4 mouse homologous genes in 3T3-L1 mature adipocytes was time-dependent, (P <0.001); (3) TNF-α intervention at 48 h (P <0.001) was inhibited by 10 ng / mL TNF-α intervention for 24 hNYGGF4 in human mature adipocytes, , The glucose uptake rate of 3T3-L1 mature adipocytes stimulated by insulin decreased more than 50% compared with the non-intervention group, and the glucose uptake stimulated by insulin was further inhibited 72 h after intervention. 【Conclusion】 TNF-α can up-regulate the homologous gene expression of NYGGF4 mouse in 3T3-L1 mature adipocytes and inhibit the insulin uptake by 3T3-L1 mature adipocytes.