Cutting edge:: Direct interaction of TLR4 with NAD(P)H oxidase 4 isozyme is essential for lipopolysaccharide-induced production of reactive oxygen species and activation of NF-κB
Cutting edge:: Direct interaction of TLR4 with NAD(P)H oxidase 4 isozyme is essential for lipopolysaccharide-induced production of reactive oxygen species and activation of NF-κB
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DOI：
10.4049/jimmunol.173.6.3589
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发表时间：
2004-09-15
影响因子：
4.4
通讯作者：
Bae, YS
中科院分区：
文献类型：
--
作者：
Park, HS;Jung, HY;Bae, YS
LPS 是革兰氏阴性菌外膜的主要成分，可被 TLR4 识别。 TLR4 与 LPS 的结合会触发各种细胞信号传导途径，包括 NF-kappaB 激活和活性氧 (ROS) 产生。在本研究中，我们提出了以下数据：在 HEK293 T 细胞中，LPS 诱导的 ROS 生成和 NF-kappaB 激活是由 TLR4 与 (NAD(P)H 氧化酶 4 (Nox) 4) 直接相互作用介导的，(NAD(P)H 氧化酶 4 (Nox) 4) 是一种与吞噬细胞 gp91(phox) (Nox2) 相关的蛋白质。酵母两种杂交和 GST Pull-down 测定表明 Nox4 的 COOH 末端区域与 TLR4 的细胞质尾部相互作用。在表达 TLR4 以及 MD2 和 CD14 的 HEK293T 细胞中转染对 Nox4 同工酶特异的小干扰 RNA 来敲低 Nox4，导致 LPS 诱导的 ROS 生成和 NF-κB 激活受到抑制。总之，这些结果表明 TLR4 与 Nox4 的直接相互作用参与 LPS 介导的 ROS 生成和 NF-κB 激活。
LPS, the primary constituent of the outer membrane of Gram-negative bacteria, is recognized by TLR4. Binding of TLR4 to LPS triggers various cell signaling pathways including NF-kappaB activation and reactive oxygen species (ROS) production. In this study, we present the data that LPS-induced ROS generation and NF-kappaB activation are mediated by a direct interaction of TLR4 with (NAD(P)H oxidase 4 (Nox) 4), a protein related to gp91(phox) (Nox2) of phagocytic cells, in HEK293 T cells. Yeast two hybrid and GST pull-down assays indicated that the COOH-terminal region of Nox4 interacted with the cytoplasmic tail of TLR4. Knockdown of Nox4 by transfection of small interference RNA specific to the Nox4 isozyme in HEK293T cells expressing TLR4 along with MD2 and CD14 resulted in inhibition of LPS-induced ROS generation and NF-kappaB activation. Taken together, these results indicate that direct interaction of TLR4 with Nox4 is involved in LPS-mediated ROS generation and NF-kappaB activation.