Role of toll-like receptors in changes in gene expression and NF-κB activation in mouse hepatocytes stimulated with lipopolysaccharide
Role of toll-like receptors in changes in gene expression and NF-κB activation in mouse hepatocytes stimulated with lipopolysaccharide
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DOI：
10.1128/iai.70.7.3433-3442.2002
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发表时间：
2002-07-01
影响因子：
3.1
通讯作者：
Billiar, TR
中科院分区：
文献类型：
--
作者：
Liu, SB;Gallo, DJ;Billiar, TR
肝脏是宿主与微生物相互作用的重要部位。尽管肝细胞已被报道对脂多糖（LPS）有反应，但LPS会改变整体基因表达，LPS刺激培养的肝细胞的机制仍不确定。用LPS培养原代小鼠肝细胞，以评估其对整体基因表达、肝脏转录因子和丝裂原活化蛋白（MAP）激酶活化的影响。DNA微阵列分析表明，LPS调节了80多个基因和表达序列标签的选择性表达。我们之前已经证明肝细胞表达CD14，这是摄取和对LPS的反应所必需的。在其他细胞中，对微生物产物的反应性需要toll样受体（TLR）及其相关附属分子的表达。逆转录酶PCR分析显示，肝细胞通过TLR9、MyD88和MD-2转录本表达TLR1，表明肝细胞表达所有已知的微生物识别分子。MAP激酶胞外信号调节激酶1/2在LPS作用下被磷酸化，tlr4缺失小鼠肝细胞的磷酸化水平较低。与对照肝细胞相比，TLR4突变或无TLR4的肝细胞中NF-kappaB的激活降低，这种缺陷通过腺病毒转导小鼠TLR4而部分恢复。因此，肝细胞通过TLR4反应途径对纳克浓度的LPS作出反应。
The liver is an important site of host-microbe interaction. Although hepatocytes have been reported to be responsive to lipopolysaccharide (LPS), the global gene expression changes by LPS and mechanism(s) by which LPS stimulates cultured hepatocytes remain uncertain. Cultures of primary mouse hepatocytes were incubated with LPS to assess its effects on the global gene expression, hepatic transcription factors, and mitogen-activated protein (MAP) kinase activation. DNA microarray analysis indicated that LPS modulates the selective expression of more than 80 genes and expressed sequence tags. We have shown previously that hepatocytes express CD14, which is required both for uptake and responsiveness to LPS. In other cells, responsiveness to microbial products requires expression of Toll-like receptors (TLR) and their associated accessory molecules. Hepatocytes expressed TLR1 through TLR9 as well as MyD88 and MD-2 transcripts, as shown by reverse transcriptase PCR analysis, indicating that hepatocytes express all known microbe recognition molecules. The MAP kinase extracellular signal-regulated kinase 1/2 was phosphorylated in response to LPS in mouse hepatocytes, and the levels of phosphorylation were lower in hepatocytes from TLR4-null mice. NF-kappaB activation was reduced in TLR4-mutant or -null hepatocytes compared to control hepatocytes, and this defect was partially restored by adenoviral transduction of mouse TLR4. Thus, hepatocytes respond to nanogram concentrations of LPS through a TLR4 response pathway.