β-Glucan modulates the lipopolysaccharide-induced innate immune response in rat mammary epithelial cells.
β-Glucan modulates the lipopolysaccharide-induced innate immune response in rat mammary epithelial cells.
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DOI：
10.1016/j.intimp.2012.12.007
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发表时间：
2013-02
影响因子：
5.6
通讯作者：
Wei Zhu;Haitian Ma;J. Miao;Guo-Qing Huang;M. Tong;S. Zou
中科院分区：
文献类型：
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作者：
Wei Zhu;Haitian Ma;J. Miao;Guo-Qing Huang;M. Tong;S. Zou
乳腺炎是由乳腺致病菌引起的一种重要疾病，在世界范围内影响着妇女和奶牛。dectin-1的β-葡聚糖结合可以诱导其自身的细胞内信号传导，并且可以介导多种细胞反应。本研究旨在探讨β-葡聚糖对脂多糖（LPS）诱导的大鼠原代乳腺上皮细胞炎症反应及相关天然免疫信号的影响。在无血清培养液中加入0、1、5、25μmol/L β-葡聚糖的DMSO溶液处理细胞12 h，然后加入10μg/mL LPS处理40 min。用BAY 11-7082预处理细胞以抑制NF-κB，然后依次暴露于5μmol/L β-葡聚糖、10μg/mL LPS、5μmol/L β-葡聚糖和10μg/mL LPS。正常对照培养物含有等体积的DMSO，同时收集。10μg/mL LPS孵育40 min后，TLR 4、MyD 88和NF-κB的表达均增加（P<0.05），TNF-α和IL-1β的分泌也增加（P <0.05），而IκB和β-酪蛋白的表达均减少（P<0.05）。不同浓度的β-葡聚糖处理12 h可激活Dectin 1/Syk，随后抑制TLR 4、MyD 88和NF-κB的表达以及TNF-α和IL-1β的分泌。而10μg/mL LPS诱导的IκB和β-酪蛋白表达则恢复。BAY 11-7082（10μmol/L）预处理2 h可部分抑制LPS诱导的NF-κB的表达，但β-葡聚糖的存在可抑制这种失活。BAY 11-7082不能同时抑制LPS诱导的大鼠乳腺上皮细胞TLR 4、MyD 88和β-葡聚糖对Dectin 1/Syk的激活。这些发现表明β-葡聚糖激活Dectin 1/Syk减弱了LPS对TLR 4/MyD 88/NF-κB的诱导，并抑制了LPS诱导的乳腺上皮细胞炎症因子，从而提供了β-葡聚糖在预防LPS诱导的乳腺上皮细胞功能障碍中的可能保护作用。
Mastitis, caused by mammary pathogenic bacteria which are frequent implications of Escherichia coli, is an important disease affecting women and dairy animals worldwide. The β-glucan binding of dectin-1 can induce its own intracellular signaling and can mediate a variety of cellular responses. This work was to investigate the effect of β-glucan on the lipopolysaccharide (LPS)-induced inflammatory response and related innate immune signaling in primary rat mammary epithelial cells. Cells were treated with serum-free medium added with a DMSO solution containing β-glucans at concentrations of 0, 1, 5, 25μmol/L for 12h, and then exposed to 10μg/mL LPS for 40min. Moreover, cells were pretreated with BAY 11-7082 to inhibit NF-κB and then successively exposed to 5μmol/L β-glucan, 10μg/mL LPS, 5μmol/L β-glucan and 10μg/mL LPS, according to the specific experimental design. Normal control cultures contained an equal volume of DMSO, which was collected at the same time. After incubating rat mammary epithelial cells for40 min with 10μg/mL LPS, TLR4, MyD88 and NF-κB expression all increased (P<0.05), as did the secretion of TNF-α and IL-1β (P<0.05), but IκB and β-casein expression both decreased (P<0.05). Treatment with different concentrations of β-glucan for 12h activated Dectin1/Syk, which subsequently suppressed TLR4, MyD88 and NF-κB expression and TNF-α and IL-1β secretion. However, it restored the IκB and β-casein expression that had been induced by the 40min incubation with 10μg/mL LPS. Pretreatment with BAY 11-7082 at 10µmol/L for 2h partially prevented NF-κB induction by LPS, but the presence of β-glucan prevented this inactivation. BAY 11-7082 could not simultaneously inhibit LPS induction of TLR4, MyD88 and β-glucan activation of Dectin1/Syk in rat mammary epithelial cells. These findings demonstrated that β-glucan activation of Dectin1/Syk attenuated LPS induction of TLR4/MyD88/NF-κB and inhibited the LPS-induced inflammation factors in mammary epithelial cells, thereby providing a possibly protective effect of β-glucan in the prevention of LPS-induced dysfunction in mammary epithelial cells.