本文關(guān)鍵詞： 胰蛋白酶抑制劑 BBI 腸道上皮細(xì)胞 炎性反應(yīng) NF-κB　出處：《中國(guó)感染控制雜志》2017年10期 　論文類型：期刊論文

【摘要】：目的探討大豆來源的胰蛋白酶抑制劑(BBI)對(duì)LPS誘導(dǎo)的腸道上皮細(xì)胞炎性因子表達(dá)的抑制作用及其機(jī)制。方法用MTT法檢測(cè)LPS和BBI對(duì)腸道上皮細(xì)胞的毒性作用。先用BBI預(yù)處理腸道上皮細(xì)胞,再用LPS刺激,采用實(shí)時(shí)熒光定量PCR檢測(cè)細(xì)胞內(nèi)炎性因子(TNF-α、IL-1β、IL-8和MCP-1)的表達(dá),通過NF-κB-luc熒光素酶質(zhì)粒和蛋白質(zhì)印跡法(Western Blot)檢測(cè)NF-κB的活性。結(jié)果 LPS最大濃度(10 000 ng/mL)和BBI最大濃度(1 000μg/mL)對(duì)細(xì)胞均無毒性作用。LPS處理可顯著地上調(diào)腸道上皮細(xì)胞炎性因子(TNF-α、IL-1β、IL-8和MCP-1)的表達(dá),其上調(diào)作用和LPS的濃度成正相關(guān);LPS處理腸道上皮細(xì)胞3 h后炎性因子表達(dá)最高,隨時(shí)間延長(zhǎng)有所下降;LPS對(duì)腸道上皮細(xì)胞炎性因子的上調(diào)作用具有劑量和時(shí)間效應(yīng);BBI預(yù)處理腸道上皮細(xì)胞6 h時(shí),BBI對(duì)LPS誘導(dǎo)腸道上皮細(xì)胞炎性因子表達(dá)的抑制作用最明顯,且具有劑量效應(yīng)。結(jié)論通過對(duì)腸道上皮細(xì)胞內(nèi)NF-κB的抑制,BBI能夠顯著地抑制LPS誘導(dǎo)炎癥因子的表達(dá)。
[Abstract]:Objective to investigate the inhibitory effect of soybean trypsin inhibitor (BBI) on the expression of inflammatory factors in intestinal epithelial cells induced by LPS and its mechanism. Methods the toxic effects of LPS and BBI on intestinal epithelial cells were detected by MTT assay. BBI was used first. Pretreatment of intestinal epithelial cells, The expression of intracellular inflammatory factor TNF- 偽, IL-1 尾, IL-8 and MCP-1 was detected by real-time fluorescence quantitative PCR (PCR) stimulated by LPS. NF- 魏 B-luc luciferase plasmid and Western blot were used to detect the activity of NF- 魏 B. Results the maximum concentration of LPS was 10 000 ng / mL) and the maximum concentration of BBI was 1 000 渭 g / mL). LPs treatment could significantly up-regulate the inflammation of intestinal epithelial cells. The expression of TNF- 偽, IL-1 尾, IL-8 and MCP-1), The up-regulation and the concentration of LPS were positively correlated with the expression of inflammatory factors in intestinal epithelial cells treated with LPS for 3 h. The upregulation of inflammatory cytokines in intestinal epithelial cells induced by LPS was the most obvious after pretreatment of intestinal epithelial cells for 6 h, and the dose and time effects of LPs on the up-regulation of inflammatory cytokines in intestinal epithelial cells were also observed. Conclusion the inhibition of NF- 魏 B in intestinal epithelial cells can significantly inhibit the expression of inflammatory factors induced by LPS.
【作者單位】： 武漢大學(xué)基礎(chǔ)醫(yī)學(xué)院;
【基金】：國(guó)家自然科學(xué)基金(No.81271334、No.81571962) 國(guó)家傳染病防治重大科技專項(xiàng)(No.2012ZX10004501-001-004)
【分類號(hào)】：R512.91

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