腹膜透析(PD)和长期暴露于PD液(PDF)引起腹膜(PM)纤维化和血管过多,导致功能性PM变性。2-脱氧葡萄糖(2-DG)通过抑制高糖酵解诱导的间皮-间质转化(MMT)而显示出作为PM抗纤维化的潜力。我们研究了2-DG与几个PDF的给药是否会影响PM的间皮和内皮屏障的通透性。2-DG的抗纤维化作用通过具有包埋的间皮(MeT-5A)或内皮(EA)的凝胶收缩测定来证实。hy926)细胞在Dianeal®2.5%(CPDF)中培养,BicaVera®2.3%(BPDF),Balance®2.3%(LPDF),添加/不添加2-DG(0.2mM),和αSMA的qPCR,CDH2基因。此外,通过监测跨膜阻力(RTM)测试2-DG对间皮和内皮细胞单层通透性的影响,FITC-葡聚糖(10,70kDa)扩散和CLDN-1至-5,ZO1,SGLT1和SGLT2基因的mRNA表达水平。MeT-5A细胞在CPDF/2-DG中的收缩性降低,伴随αSMA(0.17±0.03)和CDH2(2.92±0.29)基因表达倍数变化。αSMA的变化,在EA中发现CDH2。hy926细胞,尽管αSMA在LPDF/2-DG孵育下也降低(0.42±0.02)。总的来说,2-DG减轻了PDF诱导的间皮和内皮屏障功能的改变,如RTM所示,CLDN-1至-5、ZO1和SGLT2的葡聚糖转运和表达水平。因此,用2-DG补充PDF不仅减少了MMT,而且改善了PM间皮和内皮屏障的功能通透性特征。
Peritoneal dialysis (PD) and prolonged exposure to PD fluids (PDF) induce peritoneal membrane (PM) fibrosis and hypervascularity, leading to functional PM degeneration. 2-deoxy-glucose (2-DG) has shown potential as PM antifibrotic by inhibiting hyper-glycolysis induced mesothelial-to-mesenchymal transition (MMT). We investigated whether administration of 2-DG with several PDF affects the permeability of mesothelial and endothelial barrier of the PM. The antifibrotic effect of 2-DG was confirmed by the gel contraction assay with embedded mesothelial (MeT-5A) or endothelial (EA.hy926) cells cultured in Dianeal® 2.5 % (CPDF), BicaVera® 2.3 % (BPDF), Balance® 2.3 % (LPDF) with/without 2-DG addition (0.2 mM), and qPCR for αSMA, CDH2 genes. Moreover, 2-DG effect was tested on the permeability of monolayers of mesothelial and endothelial cells by monitoring the transmembrane resistance (RTM), FITC-dextran (10, 70 kDa) diffusion and mRNA expression levels of CLDN-1 to -5, ZO1, SGLT1, and SGLT2 genes. Contractility of MeT-5A cells in CPDF/2-DG was decreased, accompanied by αSMA (0.17 ± 0.03) and CDH2 (2.92 ± 0.29) gene expression fold changes. Changes in αSMA, CDH2 were found in EA.hy926 cells, though αSMA also decreased under LPDF/2-DG incubation (0.42 ± 0.02). Overall, 2-DG mitigated the PDF-induced alterations in mesothelial and endothelial barrier function as shown by RTM, dextran transport and expression levels of the CLDN-1 to -5, ZO1, and SGLT2. Thus, supplementation of PDF with 2-DG not only reduces MMT but also improves functional permeability characteristics of the PM mesothelial and endothelial barrier.