Determining conditions for nitric oxide synthesis in Caco-2 cells using Taguchi and factorial experimental designs
Determining conditions for nitric oxide synthesis in Caco-2 cells using Taguchi and factorial experimental designs
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DOI：
10.1016/j.ab.2008.07.013
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发表时间：
2008-10-15
影响因子：
2.9
通讯作者：
Kitts, David D.
中科院分区：
文献类型：
--
作者：
Chen, Xiu-Min;Kitts, David D.
肠道炎症与一氧化氮（NO）合成增加密切相关，这主要归因于诱导型一氧化氮合酶（iNOS）的上调。采用田口设计优化干扰素γ (ifn - γ)、肿瘤坏死因子α (tnf - α)、白细胞介素1 β (IL-1 β)、脂多糖（LPS）和肉豆酸酯phorbol myristate acetate （PMA）作为诱导物刺激Caco-2细胞NO合成。结果表明，ifn - γ是Caco-2细胞中最重要的iNOS诱导剂。使用8000 U/ml ifn - γ和0.1 μ g/ml PMA的最佳混合物处理Caco-2细胞，可协同诱导NO合成。进一步的实验采用5因素/2水平因子设计，包括Caco-2生长条件，如细胞传代、培养基组成、细胞播种时间和密度以及刺激时间。细胞播种和刺激次数显著(P
Intestinal inflammation correlates well with the increased synthesis of nitric oxide (NO), which is attributed mainly to the up-regulation of inducible nitric oxide synthase (iNOS). We optimized the use of interferon gamma (IFN-gamma), tumor necrosis factor alpha (TNF-alpha), interleukin 1 beta (IL-1 beta), lipopolysaccharide (LPS), and phorbol myristate acetate (PMA) as inducers to stimulate NO synthesis in Caco-2 cells using a Taguchi design. The results indicated that IFN-gamma was the most important inducer of iNOS in Caco-2 cells. Treating Caco-2 cells with both IFN-gamma and PMA using an optimal mixture of 8000 U/ml IFN-gamma and 0.1 mu g/ml of PMA resulted in a synergistic induction of NO synthesis. Further experiments using a 5-factor/2-level factorial design including Caco-2 growth conditions such as cell passage, Culture medium composition, cell seeding time and density, and Stimulation time were also performed. Cell seeding and stimulation times significantly (P