【摘要】：目的近年來(lái)的研究表明,人參皂苷Rd(ginsenoside Rd,G-Rd)具有神經(jīng)保護(hù)、抗炎等作用,但目前尚不清楚人參皂苷Rd在腸道上皮細(xì)胞中是否會(huì)影響炎癥反應(yīng)。本課題旨在探討人參皂苷Rd對(duì)脂多糖(lipopolysaccharide,LPS)誘導(dǎo)的人結(jié)腸癌細(xì)胞系HT-29細(xì)胞炎癥反應(yīng)的影響及作用機(jī)制。方法利用RT-qPCR(Real time-quantitative polymerase chain reaction)檢測(cè)G-Rd對(duì)LPS刺激的HT-29細(xì)胞炎癥因子IL-8的mRNA表達(dá)水平的影響;利用ELISA技術(shù)檢測(cè)G-Rd對(duì)LPS刺激的HT-29細(xì)胞分泌的炎癥因子IL-8蛋白表達(dá)水平的影響;利用免疫印跡法(Western blotting)、免疫熒光技術(shù)、EMSA檢測(cè)G-Rd對(duì)LPS誘導(dǎo)的HT-29細(xì)胞NF-κB信號(hào)通路的影響。結(jié)果1.RT-qPCR及ELISA實(shí)驗(yàn)表明:G-Rd能從mRNA水平及蛋白質(zhì)水平上抑制由LPS誘導(dǎo)的HT-29細(xì)胞中IL-8的表達(dá),且呈現(xiàn)劑量依賴關(guān)系。2.免疫熒光、免疫印跡及EMSA實(shí)驗(yàn)表明:G-Rd可下調(diào)LPS誘導(dǎo)的NF-κB信號(hào)通路的活化水平。結(jié)論G-Rd能通過(guò)抑制LPS刺激HT-29細(xì)胞分泌炎癥因子IL-8而發(fā)揮抗炎作用,其機(jī)制可能與抑制NF-κB信號(hào)通路活化有關(guān)。
[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.
【學(xué)位授予單位】：寧夏醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.35

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