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甘露糖结合凝集素对肠上皮细胞屏障功能的影响

, PP. 1091-1095

Keywords: 甘露糖结合凝集素,紧密连接蛋白,LPS

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Abstract:

目的观察甘露糖结合凝集素(mannosebindinglectin,MBL)对人结肠癌细胞株Caco-2屏障功能的影响并探讨其分子机制。方法体内实验:取14只C57小鼠,按随机数字表法分为假手术组和LPS组(经腹腔注射LPS,构建肠炎症损伤模型),通过免疫组化和Real-timePCR观察MBL的变化情况,HE染色观察肠黏膜的损伤情况。体外实验:Caco-2细胞分为对照组、空转染组和MBLshRNA组(应用MBL干扰质粒转染Caco-2细胞),采用Real-timePCR、Westernblot检测紧密连接蛋白的表达情况,并检测其跨上皮电阻(TER)。结果体内实验TER显示LPS组肠黏膜通透性(80.32±5.43)比假手术组(140.45±4.78)明显增加(P<0.01),HE染色显示LPS处理后肠黏膜完整性比假手术组降低,免疫组化和Real-timePCR检测结果显示LPS处理组MBL表达升高(P<0.05)。在体外实验中,我们成功构建MBLshRNA质粒模型,MBLshRNA组细胞中MBL的蛋白和mRNA表达[(0.34±0.09),(0.42±0.12)]较对照组及空转染组明显降低(P<0.05)。MBLshRNA组的紧密连接蛋白Occludin和Claudin-1的蛋白[(0.43±0.10)、(0.53±0.11)]和mRNA水平[(0.43±0.08)、(0.45±0.10)]明显低于其余组(P<0.05);MBLshRNA组TER值(80.33±7.64)降低较其余组明显(P<0.05)。结论下调MBL基因可减少肠上皮细胞紧密连接蛋白Occludin和Claudin-1mRNA及蛋白水平的表达,减弱肠上皮细胞的屏障保护功能。

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