Effects of lactoferrin on the expression of TLR4 signaling pathway-associated membrane molecule in LPS-activated U937 macrophages

Shen Mingyang; Wei Hong; Du Weixia; Ai Qing

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Effects of lactoferrin on the expression of TLR4 signaling pathway-associated membrane molecule in LPS-activated U937 macrophages

更新时间：2023-12-18

- Effects of lactoferrin on the expression of TLR4 signaling pathway-associated membrane molecule in LPS-activated U937 macrophages
- Journal of Chongqing Medical University Vol. 40, Issue 4, Pages: 607-611(2015)
- 作者机构：
  
  重庆医科大学附属儿童医院新生儿诊治中心、儿童发育疾病研究教育部重点实验室、儿科学重庆市重点实验室、重庆市儿童发育重大疾病诊治与预防国际科技合作基地,重庆,400014
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- Published：2015，
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Shen Mingyang, Wei Hong, Du Weixia, et al. Effects of lactoferrin on the expression of TLR4 signaling pathway-associated membrane molecule in LPS-activated U937 macrophages. [J]. Journal of Chongqing Medical University 40(4):607-611(2015)
DOI：

Shen Mingyang, Wei Hong, Du Weixia, et al. Effects of lactoferrin on the expression of TLR4 signaling pathway-associated membrane molecule in LPS-activated U937 macrophages. [J]. Journal of Chongqing Medical University 40(4):607-611(2015) DOI：

摘要

目的：研究重组人乳铁蛋白（recombinant human lactoferrin，rhLF）对脂多糖（lipopolysaccharide，LPS）激活的 U937细胞Toll样受体4（toll-like receptors 4，TLR4）信号通路相关膜分子 TLR4、CD14、MD2表达的影响。方法：经佛波酯（phorbol 12-myristate 13-acetate，PMA）诱导成熟的 U937细胞随机分为6组：对照组、LPS组（给予100 ng/ml LPS）、LPS+50 μg/ml rhLF组、LPS+100 μg/ml rhLF组、LPS+200 μg/ml rhLF组、LPS+500 μg/ml rhLF组，采用 RT-PCR和Ｗestern blot检测各组细胞 TLR4、CD14、MD2 mRNA和蛋白的表达，ELISA检测各组细胞上清液中肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）的含量。结果：LPS组U937细胞 TLR4、CD14、MD2 mRNA和蛋白的表达均明显高于对照组（P＜0.05），LPS+50 μg/ml rhLF组三者mRNA和蛋白的表达与 LPS组差异不明显（P ＞0.05），LPS+100 μg/ml rhLF组、LPS+200 μg/ml rhLF组、LPS+500 μg/ml rhLF组均明显降低了 LPS诱导的 TLR4、CD14、MD2 mRNA和蛋白的表达（P＜0.05）；LPS组 TNF-α的含量明显高于对照组（P＜0.05），LPS+50 μg/ml rhLF组、LPS+100 μg/ml rhLF组、LPS+200 μg/ml rhLF组、LPS+500 μg/ml rhLF组均明显降低了 LPS诱导的 TNF-α的产生（P＜0.05）。结论：rhLF通过抑制 TLR4介导的 LPS跨膜信号转导和炎症因子 TNF-α的分泌，对 LPS所致的脓毒血症具有潜在的治疗作用。

Abstract

Objective：To investigate the effects of recombinant human lactoferrin（rhLF） on the expression of toll-like receptors 4 （TLR4） signaling pathway-associated membrane molecule（TLR4，CD14 and MD2） in lipopolysaccharide（LPS）-activated U937 macrophages. Methods：U937 was induced to maturation by phorbol 12-myristate 13-acetate（PMA） and then divided into six groups：control group，LPS group（100 ng/ml LPS），LPS+50 μg/ml rhLF group，LPS+100 μg/ml rhLFgroup，LPS+200 μg/ml rhLF group and LPS+500 μg/ml rhLF group. The mRNA and protein expressions of TLR4，CD14 and MD2 were determined by RT-PCR and Western blot. The concentration of tumor necrosis factor-α（TNF-α） was detected by ELISA. Results：The mRNA and protein expres－sion of TLR4，CD14 and MD2 increased significantly in LPS group than in control group（P<0.05）. The mRNA and protein expression of TLR4，CD14 and MD2 had no significant change in LPS+50 μg/ml rhLF group（P >0.05），but reduced significantly in LPS+100 μg/ml rhLF group，LPS+200 μg/ml rhLF group and LPS+500 μg/ml rhLF group than in LPS group（P<0.05）. The concentration of TNF-α increased significantly in the LPS group than in the control group（P<0.05）. The levels of TNF-α reduced significantly in LPS+50 μg/ml rhLF group，LPS+100 μg/ml rhLF group，LPS+200 μg/ml rhLF group and LPS+500 μg/ml rhLF group than in LPS group（P<0.05）. Conclusion：rhLF might have a potential protective effect on LPS induced sepsis through suppressing TLR4 transmembrane signal transduction pathway and reducing the secretion of TNF-α.

关键词

乳铁蛋白脂多糖U937细胞Toll样受体4肿瘤坏死因子-α

Keywords

lactoferrinlipopolysaccharideU937 cellstoll-like receptors 4tumor necrosis factor-α

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