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当前位置: 首页 > 文章 > TGF-β1信号通路在副猪嗜血杆菌侵入3D4/21细胞中的作用 畜牧兽医学报 2018 (8) 1720-1726
Position: Home > Articles > TGF-β1 Signaling Stimulates Invasion of Haemophilus parasuis into 3D4/21 Cells Chinese Journal of Animal and Veterinary Sciences 2018 (8) 1720-1726

TGF-β1信号通路在副猪嗜血杆菌侵入3D4/21细胞中的作用

作  者：
沈懿娟;周妮妮;张健淞;王美芬;李玉峰;姜平
关键词：
副猪嗜血杆菌;纤连蛋白(Fn);α5整合素;TGF-β1;siRNA
摘  要：
本研究旨在探讨猪肺泡巨噬细胞传代细胞系3D4/21中转化生长因子β1(TGF-β1)表达和纤连蛋白(Fn)/α5整合素与副猪嗜血杆菌侵入的关系。通过吸附/侵入试验选择副猪嗜血杆菌最佳吸附/侵入条件;运用荧光定量PCR和ELISA方法检测副猪嗜血杆菌感染细胞后对TGF-β1表达的影响;用TGF-β1预处理3D4/21细胞后,用CFU检测TGF-β1表达对副猪嗜血杆菌侵入的影响;用流式细胞术检测副猪嗜血杆菌感染细胞后Fn/α5整合素表达的变化;用特异的小干扰RNA(siRNA)片段干扰Fn/α5整合素表达水平后,研究其与副猪嗜血杆菌对3D4/21吸附/侵入的影响。结果显示,副猪嗜血杆菌感染细胞后会导致TGF-β1表达增加;用重组TGF-β1预处理3D4/21细胞可以显著提高副猪嗜血杆菌的侵入水平;流式细胞术检测显示,重组TGF-β1和副猪嗜血杆菌分别处理3D4/21细胞可以增加Fn和α5整合素的表达;siRNA干扰3D4/21细胞中Fn和α5整合素的表达,细菌侵入数显著降低;然而,只有当α5整合素被干扰时,副猪嗜血杆菌的吸附数量下降。这些结果表明,TGF-β1和Fn/α5整合素的表达促进了副猪嗜血杆菌对3D4/21细胞的侵入,并在此过程中发挥重要作用。
译  名：
TGF-β1 Signaling Stimulates Invasion of Haemophilus parasuis into 3D4/21 Cells
作  者：
SHEN Yi-juan;ZHOU Ni-ni;ZHANG Jian-song;WANG Mei-fen;LI Yu-feng;JIANG Ping;Key Laboratory of Animal Bacteriology of Ministry of Agriculture,College of Veterinary Medicine,Nanjing Agricultural University;
单  位：
SHEN Yi-juan%ZHOU Ni-ni%ZHANG Jian-song%WANG Mei-fen%LI Yu-feng%JIANG Ping%Key Laboratory of Animal Bacteriology of Ministry of Agriculture,College of Veterinary Medicine,Nanjing Agricultural University
关键词：
H.parasuis;;fibronectin;;α5integrin;;TGF-β1;;siRNA
摘  要：
This study aimed to investigate the relationship between the expression of transforming growth factorβ1(TGF-β1),leading to the expression of fibronectin(Fn)/α5 integrin,and the invasion of Haemophilus parasuis(H.parasuis)in porcine alveolar macrophage cell line 3 D4/21.The optimal adherence/invasion conditions of H.parasuis to 3 D4/21 cells were determined by a series of adherence/invasion experiments.TGF-β1 expression level in the cells infected by H.parasuis was detected using Q-PCR and ELISA.Treatment of cells with recombinant TGF-β1 prior to H.parasuisinfection,CFU and flow cytometry were used to confirm the relationship between bacteria variation and the expression of TGF-β1.siRNA interference was used to down regulate the expression of Fn/α5 to confirm the relationship between the expression of Fn/α5 integrin and bacteria invasion.The H.parasuis infection resulted in the elevation of TGF-β1 expression in cells.Treatment of 3 D4/21 cells with TGF-β1 prior to infection significantly increased the invasion of H.parasuis.Flow cytometry revealed that the expression of Fn/α5 integrin was increased in 3 D4/21 cells treated with recombinant TGF-β1 and H.parasuis.Small interferingRNAs(siRNA)were used to down regulate the expression of Fn andα5 integrin in 3 D4/21 cells,which significantly decreased the invasion of H.parasuis.However,H.parasuis attachment was decreased only on cells treated with theα5 integrin siRNA,there was no corresponding decrease in attachment to cells treated with Fn siRNA.These results demonstrate that expression of TGF-β1 andα5 integrin promote the invasion of H.parasuis to 3 D4/21 cells and play an important role during this process.

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