Surfactant protein-D attenuates the lipopolysaccharide-induced inflammation in human intestinal cells overexpressing toll-like receptor 4
Surfactant protein-D attenuates the lipopolysaccharide-induced inflammation in human intestinal cells overexpressing toll-like receptor 4
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DOI：
10.1007/s00383-015-3812-y
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发表时间：
2016-01-01
影响因子：
1.8
通讯作者：
Okuyama, Hiroomi
中科院分区：
文献类型：
--
作者：
Saka, Ryuta;Wakimoto, Tetsu;Okuyama, Hiroomi
坏死性小肠结肠炎 (NEC) 是早产儿的一种破坏性炎症性疾病，可能依赖于未成熟肠道中 Toll 样受体 4 (TLR4) 的过度表达。表面活性蛋白 (SP)-D 是集合素家族的一员，在先天免疫中发挥着重要作用，特别是在气道中。尽管 SP-D 也存在于肠道中，但对其功能知之甚少。本研究探讨SP-D是否可以减弱TLR4过表达胚胎肠细胞的炎症反应。所有实验程序均使用最初源自人胚胎肠的人肠细胞系INT407进行。据报道，NEC 患者中血小板激活因子 (PAF) 升高，该因子被用来诱导人胚胎肠细胞系 INT407 中 TLR4 过度表达。使用定量实时 PCR 测量 TLR4 表达。通过白细胞介素 8 (IL-8) 释放（以 pg/ml 为单位）的酶联免疫吸附测定（ELISA）评估 SP-D 预处理（20 A mu g/ml）后对 PAF（5 A muM）、TLR4 激动剂脂多糖（LPS，100 ng/ml）、PAF + LPS 和 PAF + LPS 的炎症反应。TLR4 mRNA 的表达（平均值 +/- A SD）为PAF 上调 (369 % +/- A 28 %, p < 0.001)。 PAF + LPS 刺激导致 IL-8 释放量 (1959.3 +/- A 52.3) 高于对照 (141.2 +/- A 12.4)、LPS (167.3 +/- A 65.8) 或 PAF (1527.2 +/- A 129.4) 治疗 (p < 0.05)。 SP-D 预处理减弱了 PAF + LPS 的释放 (1590.1 +/- A 319.3) (1161.6 +/- A 131.6; p < 0.05)。SP-D 减弱了 TLR4 过表达肠道细胞中 LPS 诱导的 IL-8 产生，表明 SP-D 可能对早产儿 NEC 的发展具有保护作用。
Necrotizing enterocolitis (NEC) is a devastating inflammatory disease of preterm infants that may depend on overexpression of toll-like receptor-4 (TLR4) in the immature intestine. Surfactant protein (SP)-D is a member of the collectin family and plays an important role in innate immunity, particularly in the airways. Although SP-D also exists in the intestines, little is known about its function. This study investigated whether SP-D can attenuate the inflammatory response of TLR4-overexpressing embryonal intestinal cells.All experimental procedures were performed using the human intestinal cell line INT407 originally derived from human embryonal intestines. Platelet-activating factor (PAF), reported to be elevated in NEC patients, was used to induce TLR4 overexpression in the human embryonal intestinal cell line INT407. TLR4 expression was measured using quantitative real-time PCR. Inflammatory responses to PAF (5 A mu M), the TLR4 agonist lipopolysaccharide (LPS, 100 ng/ml), PAF + LPS, and PAF + LPS following SP-D pretreatment (20 A mu g/ml) were assessed by enzyme-linked immunosorbent assay (ELISA) of interleukin-8 (IL-8) release (in pg/ml).Expression of TLR4 mRNA (mean +/- A SD) was upregulated by PAF (369 % +/- A 28 %, p < 0.001). Stimulation with PAF + LPS resulted in higher IL-8 release (1959.3 +/- A 52.3) than control (141.2 +/- A 12.4), LPS (167.3 +/- A 65.8), or PAF (1527.2 +/- A 129.4) treatment (p < 0.05). Release in response to PAF + LPS (1590.1 +/- A 319.3) was attenuated by SP-D pretreatment (1161.6 +/- A 131.6; p < 0.05).SP-D attenuates LPS-induced IL-8 production in TLR4-overexpressing intestinal cells, suggesting that SP-D may have a protective effect in the development of NEC in preterm infants.