Evidence for the presence of Toll-like receptor 4 system in the human endometrium
Evidence for the presence of Toll-like receptor 4 system in the human endometrium
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DOI：
10.1210/jc.2004-0241
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发表时间：
2005-01-01
影响因子：
5.8
通讯作者：
Taketani, Y
中科院分区：
文献类型：
--
作者：
Hirata, T;Osuga, Y;Taketani, Y
我们研究了toll样受体（TLR） 4是否在人子宫内膜中起作用。采用RT-PCR和原位杂交技术检测TLR4 mRNA在子宫内膜上皮细胞（EECs）和基质细胞（ESCs）中的表达。Western blotting分析显示TLR4蛋白在两种细胞群中均有表达。脂多糖（LPS）的作用是通过TLR4介导的，其处理显著增加了培养的ESCs的IL-8分泌，并呈剂量依赖性。LPS的刺激作用被加入TLR4和CD14的中和抗体所抑制。LPS还刺激ESCs中核因子- kappab的核易位，这也被这些抗体抑制。另一方面，LPS没有刺激培养的EECs分泌IL-8。然而，在可溶性CD14存在的情况下，LPS刺激EECs分泌IL-8。流式细胞分析显示，CD14在ESCs细胞表面表达，而在EECs细胞表面不表达。此外，免疫组化分析显示，内皮细胞中有CD14染色，而内皮细胞中没有。预处理干扰素- γ (ifn - γ)可增强lps诱导的内皮细胞IL-8分泌。ifn - γ增加了tlr4mrna的表达。它还增加了CD14、MD2和MyD88 mRNA的数量，这些mRNA是与TLR4一起识别LPS所必需的。综上所述，我们证明了ESCs和EECs都表达TLR4，并通过TLR4响应LPS。我们进一步表明，EECs而非ESCs需要可溶性CD14来激活TLR4。有趣的是，ifn - γ，一种抗感染细胞因子，被发现激活内皮细胞中的TLR4系统。总之，这些结果表明TLR4系统可能代表了人类子宫内膜的局部免疫，但在ESCs和EECs中TLR4的作用模式不同。
We investigated whether Toll-like receptor (TLR) 4 is at work in the human endometrium. The expression of TLR4 mRNA in endometrial epithelial cells (EECs) and stromal cells (ESCs) was detected by RT-PCR and in situ hybridization. Western blotting analysis revealed the TLR4 protein expression in both cell populations. Treatment of lipopolysaccharide (LPS), the actions of which are mediated through TLR4, significantly increased IL-8 secretion from cultured ESCs in a dose-dependent fashion. The stimulatory effect of LPS was inhibited by the addition of neutralizing antibodies for TLR4 and CD14. LPS also stimulated nuclear translocation of nuclear factor-kappaB in ESCs, which was also inhibited by these antibodies. On the other hand, LPS did not stimulate IL-8 secretion in cultured EECs. However, LPS stimulated IL-8 secretion from EECs in the presence of soluble CD14. Flow cytometric analysis revealed that CD14 was expressed on the cell surface of ESCs but not EECs. In addition, immunohistochemical analysis showed that CD14 was stained in ESCs but not EECs. Pretreatment of interferon-gamma (IFN-gamma) enhanced LPS-induced IL-8 secretion from ESCs. IFN-gamma increased the expression of TLR4 mRNA. It also increased the amounts of mRNA for CD14, MD2, and MyD88, which are needed for LPS recognition in concert with TLR4. In summary, we demonstrated that both ESCs and EECs express TLR4 and respond to LPS through TLR4. We further showed that EECs, not ESCs, required soluble CD14 for TLR4 activation. Interestingly, IFN-gamma, an antiinfectious cytokine, was found to activate the TLR4 system in ESCs. Altogether, the results imply that the TLR4 system might represent local immunity in the human endometrium with differential modes of TLR4 actions between ESCs and EECs.