Imbalance of Wnt/Dkk negative feedback promotes persistent activation of pancreatic stellate cells in chronic pancreatitis.
Imbalance of Wnt/Dkk negative feedback promotes persistent activation of pancreatic stellate cells in chronic pancreatitis.
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Wnt/Dkk负反馈失衡促进慢性胰腺炎胰腺星状细胞持续激活
DOI：
10.1371/journal.pone.0095145
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Hu G
中科院分区：
文献类型：
--
作者：
Hu Y;Wan R;Yu G;Shen J;Ni J;Yin G;Xing M;Chen C;Fan Y;Xiao W;Xu G;Wang X;Hu G
胰腺星状细胞（PSC）的持续激活在慢性胰腺炎（CP）相关纤维化中的作用正日益被认识。近年来的研究表明，Wnt信号通路参与了多器官纤维化的发生发展，但特异性Wnt在胰腺纤维化中的作用尚不清楚。我们研究了Wnt信号在CP PSC活化过程中的作用以及β-catenin抑制和Dickkopf相关蛋白1（Dkk 1）恢复对PSC表型的影响。通过反复注射雨蛙肽诱导小鼠CP，分离并体外激活小鼠PSC。通过定量RT-PCR和蛋白质印迹法分析Wnts、β-catenin、分泌性卷曲相关蛋白（sFRPs）和Dkks的表达。通过免疫荧光和Western blot检测细胞核β-catenin的表达来检查经典的Wnt信号通路。采用流式细胞术、免疫荧光、免疫细胞化学和CCK-8分析法检测重组小鼠Dkk-1（rmDkk-1）对细胞增殖和凋亡的影响。采用定量RT-PCR和Western blotting方法检测不同浓度的rmDkk-1或针对β-catenin的siRNA处理后PSC中β-catenin、胶原1 α1、TGFβRII、PDGFRβ和α-SMA的表达。Wnt 2是唯一在PSC激活后表达显著上调的Wnt，CP小鼠胰腺中Wnt 2和β-catenin蛋白水平显著升高，而Dkk-1表达明显降低。rmDkk-1通过下调PDGFRβ和TGFβRII的表达，抑制β-catenin的核转位，抑制PSC的增殖和细胞外基质的生成。Dkk-1表达上调增加培养的PSC中的凋亡。这些结果表明Wnt信号可能介导PSC激活的促纤维化作用，并且Wnt 2/Dkk-1可能是CP的潜在治疗靶点。
The role of persistent activation of pancreatic stellate cells (PSCs) in the fibrosis associated with chronic pancreatitis (CP) is increasingly being recognized. Recent studies have shown that Wnt signaling is involved in the development of fibrosis in multiple organs, however, the role of specific Wnts in pancreatic fibrosis remains unknown. We investigated the role of Wnt signaling during PSC activation in CP and the effect of β-catenin inhibition and Dickkopf-related protein 1 (Dkk1) restoration on the phenotype of PSCs. CP was induced in mice by repetitive caerulein injection and mouse PSCs were isolated and activated in vitro. The expression of Wnts, β-catenin, secreted frizzled-related proteins (sFRPs) and Dkks was analyzed by quantitative RT-PCR and western blotting. The canonical Wnt signaling pathway was examined by immunofluorescence and western blot detection of nuclear β-catenin expression. The effect of recombinant mouse Dkk-1 (rmDkk-1) on cell proliferation and apoptosis was assessed by flow cytometry, immunofluorescence, immunocytochemistry and Cell Counting Kit-8 (CCK-8) analysis. The expression of β-catenin, collagen1α1, TGFβRII, PDGFRβ and α-SMA in PSCs treated with different concentrations of rmDkk-1 or siRNA against β-catenin was determined by quantitative RT-PCR and western blotting. Wnt2 was the only Wnt whose expression was significantly upregulated in response to PSC activation, and Wnt2 and β-catenin protein levels were significantly increased in the pancreas of CP mice, whereas Dkk-1 expression was evidently decreased. Nuclear β-catenin levels were markedly increased in activated PSCs, and rmDkk-1 suppressed the nuclear translocation of β-catenin and the proliferation and extracellular matrix production of PSCs through the downregulation of PDGFRβ and TGFβRII. Upregulation of Dkk-1 expression increased apoptosis in cultured PSCs. These results indicate that Wnt signaling may mediate the profibrotic effect of PSC activation, and Wnt2/Dkk-1 could be potential therapeutic targets for CP.
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影响因子：
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作者：
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通讯作者：
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