人参皂苷Rd对人结肠癌细胞系表达炎症因子的影响

发布时间：2018-08-26 13:17

【摘要】：目的近年来的研究表明,人参皂苷Rd(ginsenoside Rd,G-Rd)具有神经保护、抗炎等作用,但目前尚不清楚人参皂苷Rd在肠道上皮细胞中是否会影响炎症反应。本课题旨在探讨人参皂苷Rd对脂多糖(lipopolysaccharide,LPS)诱导的人结肠癌细胞系HT-29细胞炎症反应的影响及作用机制。方法利用RT-qPCR(Real time-quantitative polymerase chain reaction)检测G-Rd对LPS刺激的HT-29细胞炎症因子IL-8的mRNA表达水平的影响;利用ELISA技术检测G-Rd对LPS刺激的HT-29细胞分泌的炎症因子IL-8蛋白表达水平的影响;利用免疫印迹法(Western blotting)、免疫荧光技术、EMSA检测G-Rd对LPS诱导的HT-29细胞NF-κB信号通路的影响。结果1.RT-qPCR及ELISA实验表明:G-Rd能从mRNA水平及蛋白质水平上抑制由LPS诱导的HT-29细胞中IL-8的表达,且呈现剂量依赖关系。2.免疫荧光、免疫印迹及EMSA实验表明:G-Rd可下调LPS诱导的NF-κB信号通路的活化水平。结论G-Rd能通过抑制LPS刺激HT-29细胞分泌炎症因子IL-8而发挥抗炎作用,其机制可能与抑制NF-κB信号通路活化有关。
[Abstract]:Objective recent studies have shown that ginsenoside Rd (ginsenoside Rd,G-Rd has neuroprotective and anti-inflammatory effects, but it is not clear whether ginsenoside Rd affects inflammation in intestinal epithelial cells. The aim of this study was to investigate the effect and mechanism of ginsenoside Rd on lipopolysaccharide (lipopolysaccharide,LPS) -induced inflammatory response of human colon cancer cell line HT-29. Methods RT-qPCR (Real time-quantitative polymerase chain reaction) was used to detect the effect of G-Rd on the mRNA expression of IL-8 in HT-29 cells stimulated by LPS, and ELISA technique was used to detect the effect of G-Rd on the expression of IL-8 protein, which was secreted by LPS stimulated HT-29 cells. The effect of G-Rd on NF- 魏 B signaling pathway in HT-29 cells induced by LPS was detected by immunoblotting (Western blotting), immunofluorescence technique. Results 1.RT-qPCR and ELISA experiments showed that G Rd could inhibit the expression of IL-8 in HT-29 cells induced by LPS at the level of mRNA and protein in a dose-dependent manner. Immunofluorescence, immunoblotting and EMSA assay showed that the activation level of NF- 魏 B signaling pathway induced by LPS could be down-regulated by 1: G Rd. Conclusion G-Rd can play an anti-inflammatory effect by inhibiting the secretion of inflammatory factor IL-8 by HT-29 cells stimulated by LPS. The mechanism may be related to the inhibition of activation of NF- 魏 B signaling pathway.
【学位授予单位】：宁夏医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.35

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