BBI抑制LPS诱导的肠道上皮细胞炎性因子的表达
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霍文哲

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R574

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国家自然科学基金(No. 81271334、No.81571962);国家传染病防治重大科技专项(No. 2012ZX10004501001004)


BBI inhibits LPSinduced expression of inflammatory cytokines in intestinal epithelial cells
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    摘要:

    目的探讨大豆来源的胰蛋白酶抑制剂(BBI)对LPS诱导的肠道上皮细胞炎性因子表达的抑制作用及其机制。方法用MTT法检测LPS和BBI对肠道上皮细胞的毒性作用。先用BBI预处理肠道上皮细胞,再用LPS刺激,采用实时荧光定量PCR检测细胞内炎性因子(TNFα、IL1β、IL8和MCP1)的表达,通过NFκBluc荧光素酶质粒和蛋白质印迹法(Western Blot)检测NFκB的活性。 结果LPS最大浓度(10 000 ng/mL)和BBI最大浓度(1 000 μg/mL)对细胞均无毒性作用。LPS处理可显著地上调肠道上皮细胞炎性因子(TNFα、IL1β、IL8和MCP1)的表达,其上调作用和LPS的浓度成正相关;LPS处理肠道上皮细胞3 h后炎性因子表达最高,随时间延长有所下降;LPS对肠道上皮细胞炎性因子的上调作用具有剂量和时间效应;BBI预处理肠道上皮细胞6 h时,BBI对LPS诱导肠道上皮细胞炎性因子表达的抑制作用最明显,且具有剂量效应。结论通过对肠道上皮细胞内NFκB的抑制,BBI能够显著地抑制LPS诱导炎症因子的表达。

    Abstract:

    ObjectiveTo investigate the inhibitory effect and mechanism of soybeanderived BowmanBirk inhibitor (BBI) on LPSinduced expression of inflammatory cytokines in intestinal epithelial cells.MethodsCytotoxicity effect of LPS and BBI on intestinal epithelial cells was analyzed by MTT assay. Intestinal epithelial cells were pretreated with BBI, followed by LPS stimulation, expression of inflammatory cytokines(TNFα, IL1β, IL8, and MCP1) was detected by quantitative realtime polymerase chain reaction; the activation of NFκB was measured by pNFκBluc system and Western Blot.ResultsThe maximum concentration of LPS(10 000 ng/mL) and  BBI(1 000 μg/mL)had no cytotoxicity effect on intestinal epithelial cells. LPS could potently upregulate the expression of inflammatory cytokines(TNFα, IL1β, IL8, and MCP1), the upregulation was positively correlated to the concentration of LPS; LPSinduced expression of inflammatory cytokines in intestinal epithelial cells could achieve the highest level, then decreased over time. The upregulation of LPS on inflammatory cytokines in intestinal epithelial cells had dosetime effect; when intestinal epithelial cells were pretreated by BBI for 6 hours, the inhibitory effect of BBI on LPSinduced expression of inflammatory cytokines in intestinal epithelial cells was most obvious, and had dosetime effect.ConclusionBBI can potently inhibit LPSinduced expression of  inflammatory cytokines through inhibiting NFκB in intestinal epithelial cells.

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古俊,郭乐,刘金彪,等. BBI抑制LPS诱导的肠道上皮细胞炎性因子的表达[J]. 中国感染控制杂志,2017,16(10):893-898. DOI:10.3969/j. issn.1671-9638.2017.10.001.
GU Jun, GUO Le, LIU Jinbiao, et al. BBI inhibits LPSinduced expression of inflammatory cytokines in intestinal epithelial cells[J]. Chin J Infect Control, 2017,16(10):893-898. DOI:10.3969/j. issn.1671-9638.2017.10.001.

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  • 收稿日期:2017-07-13
  • 最后修改日期:2017-08-02
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  • 在线发布日期: 2017-09-30
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