溃结灵对Caco-2炎症细胞模型核因子-κB p65 DNA结合活性的影响

Effect of Kuijieling Decoction on DNA-Binding Activity of Nuclear Factor Kappa B p65 in Caco-2 Cell Model of Inflammation

【目的】观察溃结灵含药血清对Caco-2炎症细胞模型核因子-κB(NF-κB)p65蛋白DNA结合活性的影响。【方法】采用不同剂量含药血清干预Caco-2细胞,并以促炎细胞因子[肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)]诱导Caco-2细胞建立炎症细胞模型,收集细胞。采用Trans AM核转录因子活性检测试剂盒检测细胞核蛋白NF-κB p65的含量(活性)。【结果】TNF-ɑ与IL-1β诱导的模型组细胞核蛋白NF-κB p65的含量显著高于正常对照组(P﹤0.01);溃结灵高、中剂量组,蛋白酶抑制剂组和阳性药(柳氮磺胺吡啶)组细胞核蛋白NF-κB p65的含量均显著降低,与模型组比较差异有统计学意义(P﹤0.01)。【结论】溃结灵含药血清对Caco-2炎症细胞模型细胞核蛋白NF-κB p65 DNA结合活性具有一定的下调作用。

Objective To observe the effects of serum containing Kuijieling Decoction（KD） on DNA-binding activity of nuclear factor Kappa B p65（NF-κBp65） protein in Caco-2 cell model of inflammation.Methods The Caco-2 cell model of inflammation was induced by tumor necrosis factor alpha（TNF-α）or interleukin 1 beta（IL-1β）,and then were co-cultured with blank serum,proteinase inhibitor Bortezomib,salicylazosulfapyridine（SASP） or different dosages of KD-containing serum.The content of nuclear protein NF-κB p65 was determined by Trans AM nuclear transcription factor assay kit.Results The content of nuclear protein NF-κB p65 was significantly higher in the model cells induced by TNF-ɑ or IL-1β than that in the normal control group（P0.01）.The content of nuclear protein NF-κB p65 was significantly lower in high-and moderate-dose KD groups,Bortezomib group and SASP group than that in the model group（P0.01）.Conclusion Serum containing KD can down-regulate the DNA-binding activity of nuclear protein NF-κB p65 in Caco-2 cell model of inflammation.