Role of transient receptor potential vanilloid 2 in LPS-induced cytokine production in macrophages
Role of transient receptor potential vanilloid 2 in LPS-induced cytokine production in macrophages
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DOI：
10.1016/j.bbrc.2010.06.082
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发表时间：
2010-07-23
影响因子：
3.1
通讯作者：
Furukawa, Tetsushi
中科院分区：
文献类型：
--
作者：
Yamashiro, Kenji;Sasano, Tetsuo;Furukawa, Tetsushi
有相当多的证据表明，细胞内Ca2+作为第二信使参与tlr4依赖性信号传导。然而，细胞内游离Ca2+浓度（[Ca2+](i)）如何响应LPS而增加，以及它们如何影响细胞因子的产生，目前尚不清楚。在这里，我们研究了瞬时受体电位（TRP）的作用，这是不可兴奋细胞中主要的Ca2+渗透途径，在lps诱导的巨噬细胞中产生细胞因子。药理学实验表明，TRPV家族成员参与了lps诱导的RAW264巨噬细胞TNF α和IL-6的产生，而TRPC和TRPM家族成员均未参与。RT-PCR和免疫印迹分析显示，TRPV2是TRPV家族中唯一在巨噬细胞中表达的成员。TRPV2的ShRNA抑制了lps诱导的TNF α和IL-6的产生以及I κ B α的降解。使用BAPTA/AM和EGTA和Ca2+成像的实验表明，lps诱导的[Ca2+]增加(i)涉及trpv2介导的细胞内和细胞外Ca2+动员。BAPTA/AM可以抑制lps诱导的TNF - α和IL-6的产生，而EGTA只能部分抑制lps诱导的IL-6的产生，而不能抑制TNF - α的产生。这些数据表明，TRPV2参与了lps诱导的Ca2+从细胞内Ca2+储存和细胞外Ca2+的动员。除了通过ip3受体动员Ca2+外，trpv2介导的细胞内Ca2+动员参与NF kappa b依赖性TNFa和IL-6的表达，而细胞外Ca2+进入参与NF kappa b非依赖性IL-6的产生。(C) 2010爱思唯尔公司版权所有。
There is considerable evidence indicating that intracellular Ca2+ participates as a second messenger in TLR4-dependent signaling. However, how intracellular free Ca2+ concentrations ([Ca2+](i)) is increased in response to LPS and how they affect cytokine production are poorly understood. Here we examined the role of transient receptor potential (TRP), a major Ca2+ permeation pathway in non-excitable cells, in the LPS-induced cytokine production in macrophages. Pharmacologic experiments suggested that TRPV family members, but neither TRPC nor TRPM family members, are involved in the LPS-induced TNF alpha and IL-6 production in RAW264 macrophages. RT-PCR and immunoblot analyses showed that TRPV2 is the sole member of TRPV family expressed in macrophages. ShRNA against TRPV2 inhibited the LPS-induced TNF alpha and IL-6 production as well as I kappa B alpha degradation. Experiments using BAPTA/AM and EGTA, and Ca2+ imaging suggested that the LPS-induced increase in [Ca2+](i) involves both the TRPV2-mediated intracellular and extracellular Ca2+ mobilizations. BAPTA/AM abolished LPS-induced TNF alpha and IL-6 production, while EGTA only partially suppressed LPS-induced IL-6 production, but not TNF alpha production. These data indicate that TRPV2 is involved in the LPS-induced Ca2+ mobilization from intracellular Ca2+ store and extracellular Ca2+. In addition to Ca2+ mobilization through the IP3-receptor, TRPV2-mediated intracellular Ca2+ mobilization is involved in NF kappa B-dependent TNFa and IL-6 expression, while extracellular Ca2+ entry is involved in NF kappa B-independent IL-6 production. (C) 2010 Elsevier Inc. All rights reserved.