野生蒙古口蘑多糖通过STAT3和HIF-1α通路对LPS诱导的巨噬细胞发挥抗炎调节作用
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Anti-Inflammatory Effect of Wild Tricholoma mongolicum Polysaccharides on LPS-Induced RAW264.7 Cells via STAT3 and HIF-1α Pathways
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    摘要:

    研究蒙古口蘑多糖提取物对脂多糖(LPS)诱导小鼠单核巨噬细胞(RAW264.7细胞)炎症保护作用的机理。用不同预量质量浓度的蒙古口蘑多糖处理RAW264.7细胞,通过CCK-8法检测细胞活性;Griess法测定NO的产生量;酶联免疫吸附测定肿瘤坏死因子-a(TNF-α),白介素1?茁(IL-1?茁);实时定量检测缺氧诱导因子1α(HIF-1α)、诱导型一氧化氮和酶(iNOS)、核因子κB(NF-κB)、信号转导和转录激活因子3(STAT3)和环氧化酶2(COX-2)的mRNA表达量;免疫印迹检测HIF-1α、NF-κB、STAT3和COX-2蛋白质的表达量。结果表明,蒙古口蘑多糖提取物作用RAW264.7细胞的最大剂量为100 μg/mL;与LPS模型组相比,蒙古口蘑多糖提取物质量浓度0.01、0.1、1、10 μg/mL均能有效抑制NO(抑制率分别是38.21%、64.17%、58.16%和79.42%)、TNF-α(抑制率分别为27.91%、36.31%、36.14%、45.09%)和IL-1?茁(抑制率分别为47.75%、58.47%、60.21%、64.55%)的产生;在质量浓度为10 μg/mL的蒙古口蘑多糖提取物作用下,除NF-κB 外,HIF-1α、STAT3、COX-2和iNOS的mRNA的表达量均降低;免疫印迹的结果也显示,除NF-κB 外,HIF-1α、NF-κB、STAT3及COX-2的蛋白质的表达量均有不同程度的降低。蒙古口蘑多糖提取物对脂多糖(LPS)诱导的RAW264.7细胞具有保护作用,其抗炎的机制主要通过JAK-STAT3及HIF-1α信号通路,而不主要通过NF-κB信号通路发挥作用。

    Abstract:

    This article aimed to study the anti-inflammatory mechanism of Tricholoma mongolicum polysaccharide extract in LPS-treated RAW264.7 cells. The RAW264.7 cells were first treated with different concentrations of Tricholoma mongolicum polysaccharides, and the viability was determined by CCK-8. The release of NO was measured by Griess, and TNF-alpha and IL-1 beta were examined by ELISA. The expression levels of HIF-1α,NF-κB,STAT3,COX-2 and iNOS mRNAs and proteins were tested by qPCR and western blot. The results showed that the maximum safety dose of Tricholoma mongolicum polysaccharide extract was less than 100 μg/mL. Compared with the LPS model group, Tricholoma mongolicum polysaccharide extract at the concentrations of 0.01, 0.1, 1, 10 μg/mL could inhibit the production of NO,TNF-α and IL-1β. The inhibition rates of NO were 38.21%, 64.17%, 58.16%, 79.42%, respectively; the inhibition rates of TNF-α were 27.91%, 36.31%, 36.14%, 45.09%, respectively; the inhibition rates of IL-1?茁 were 47.75%, 58.47%, 60.21%, 64.55%, respectively. Tricholoma mongolicum polysaccharide extract at the concentration of 10 ?滋g/mL could down-regulate the mRNAs and proteins expression levels of HIF-1α, iNOS, STAT3 and COX-2. Therefore, anti-inflammatory effect of wild Tricholoma mongolicum polysaccharides on LPS-induced RAW264.7 cells may be exerted via STAT3 and HIF-1α pathways.

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于传宗,张凤兰,慕宗杰,李子钦,孙峰成,郝丽珍.野生蒙古口蘑多糖通过STAT3和HIF-1α通路对LPS诱导的巨噬细胞发挥抗炎调节作用[J].食品与生物技术学报,2020,39(6):93-98.

YU Chuanzong, ZHANG Fenglan, MU Zongjie, LI Ziqin, SUN Fengcheng, HAO Lizhen. Anti-Inflammatory Effect of Wild Tricholoma mongolicum Polysaccharides on LPS-Induced RAW264.7 Cells via STAT3 and HIF-1α Pathways[J]. Journal of Food Science and Biotechnology,2020,39(6):93-98.

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  • 在线发布日期: 2020-10-20
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