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脂联素 2 缺乏小鼠在正常和炎症条件下的蛋白质组学分析。

Proteomic profiling in Lipocalin 2 deficient mice under normal and inflammatory conditions.

机构信息

Institute of Pathology, RWTH University Hospital Aachen, Germany.

出版信息

J Proteomics. 2013 Jan 14;78:188-96. doi: 10.1016/j.jprot.2012.11.021. Epub 2012 Dec 3.

Abstract

Lipocalin 2 (LCN2) belongs to the superfamily of lipocalins which represent a group of small secreted proteins classified as extracellular transport proteins expressed in many tissues. LCN2 is strongly increased in experimental models of acute and chronic liver injuries. To investigate the function of LCN2 in normal liver homeostasis and under conditions of inflammatory liver injury, we comparatively analyzed hepatic extracts taken from Lcn2-deficient and wild type mice under basal conditions and after stimulation with lipopolysaccharides. Liver was chemically and mechanically lysed and extracts were subjected to 2-D-DIGE after minimal labeling (G200 and G300 dyes) using an appropriate internal standard (G100). Afterwards MALDI TOF MS and MS/MS were used to identify differentially expressed proteins. Proteins that were identified to be differentially expressed include for example the chloride intracellular channel protein 4 (CLIC4), aminoacylase 1 and transketolase. The altered expression of respective genes was confirmed by Western blot analysis and further validated by quantitative real time PCR. Altogether, the complex expression alterations in mice lacking LCN2 under normal conditions and after exposure to inflammatory stimuli reveal that LCN2 has essential function in liver homeostasis and in the onset of inflammatory responses in which LCN2 expression dramatically increases.

摘要

脂质运载蛋白 2(LCN2)属于脂质运载蛋白超家族,该超家族代表了一组小分泌蛋白,被归类为在许多组织中表达的细胞外转运蛋白。LCN2 在急性和慢性肝损伤的实验模型中强烈增加。为了研究 LCN2 在正常肝脏稳态和炎症性肝损伤条件下的功能,我们比较分析了基础条件下和用脂多糖刺激后 Lcn2 缺陷型和野生型小鼠的肝提取物。用化学和机械方法裂解肝脏,并用适当的内标(G100)进行最小标记(G200 和 G300 染料)后,进行 2-D-DIGE。然后使用 MALDI-TOF MS 和 MS/MS 鉴定差异表达的蛋白质。鉴定为差异表达的蛋白质包括氯离子细胞内通道蛋白 4(CLIC4)、氨基酰化酶 1 和转酮醇酶。通过 Western blot 分析证实了相应基因的改变表达,并通过定量实时 PCR 进一步验证。总之,缺乏 LCN2 的小鼠在正常条件下和暴露于炎症刺激下的复杂表达改变表明,LCN2 在肝脏稳态和炎症反应的发生中具有重要功能,其中 LCN2 的表达显著增加。

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