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[目的]研究紫锥菊多糖(EPS)在内毒素(LPS)损伤小肠上皮细胞(IEC-6)时对肿瘤坏死因子(TNF)表达的影响,以探讨EPS对损伤细胞的作用机制。[方法]采用TRIzon试剂提取总RNA,RT-PCR扩增TNF-αm RNA,琼脂糖凝胶电泳,并进行电泳及图像分析。[结果]50μg/ml EPS可以部分抑制LPS刺激IEC-6产生的TNF-αm RNA水平,而200、500μg/ml EPS随着浓度的增加,其抑制TNF-αm RNA的水平逐渐增加;将IEC-6分别用50、100、200及500μg/ml EPS预处理24 h,然后用10μg/ml LPS刺激达1、4 h,采用RT-PCR方法分析得,LPS诱导TNF-αm RNA表达被EPS有效地抑制,4h的抑制率高于1 h的抑制率。[结论]EPS通过抑制LPS刺激细胞分泌TNF-αm RNA的产生而起到肠道粘膜的保护作用,且EPS对这种抑制作用具有浓度及时间依赖性。
[Objective] To investigate the effect of Echinacea on the expression of tumor necrosis factor (TNF) in the endotoxin (LPS) -induced intestinal epithelial cells (IEC-6) to explore the mechanism of EPS on injured cells. [Method] Total RNA was extracted by TRIzon reagent, TNF-αm RNA was amplified by RT-PCR and electrophoresed on agarose gel. Electrophoresis and image analysis were performed. [Results] 50μg / ml EPS could partially inhibit the level of TNF-αmRNA produced by IEC-6 stimulated by LPS, while the level of TNF-αmRNA increased gradually with 200,500μg / ml EPS. 6 were pretreated with 50, 100, 200 and 500μg / ml EPS for 24 h, then stimulated with 10μg / ml LPS for 1 and 4 h respectively. The expression of TNF-αmRNA induced by LPS was analyzed by RT-PCR. Inhibition, 4h inhibition rate higher than 1 h inhibition rate. [Conclusion] EPS can protect the intestinal mucosa by inhibiting the production of TNF-αmRNA secreted by LPS, and the inhibitory effect of EPS on the concentration and time-dependent manner.