Recent animal studies, as well as quantitative sodium MRI observations on humans demonstrated that remarkable amounts of sodium can be stored in the skin. It is also known that excess sodium in the tissues leads to inflammation in various organs, but its role in dermal pathophysiology has not been elucidated. Therefore, our aim was to study the effect of dietary salt loading on inflammatory process and related extracellular matrix (ECM) remodeling in the skin. To investigate the effect of high salt consumption on inflammation and ECM production in the skin mice were kept on normal (NSD) or high salt (HSD) diet and then dermatitis was induced with imiquimod (IMQ) treatment. The effect of high salt concentration on dermal fibroblasts (DF) and peripheral blood mononuclear cells (PBMC) was also investigated in vitro. The HSD resulted in increased sodium content in the skin of mice. Inflammatory cytokine Il17 expression was elevated in the skin of HSD mice. Expression of anti-inflammatory Il10 and Il13 decreased in the skin of HSD or HSD IMQ mice. The fibroblast marker Acta2 and ECM component Fn and Col1a1 decreased in HSD IMQ mice. Expression of ECM remodeling related Pdgfb and activation phosphorylated (p)-SMAD2/3 was lower in HSD IMQ mice. In PBMCs, production of IL10, IL13 and PDGFB was reduced due to high salt loading. In cultured DFs high salt concentration resulted in decreased cell motility and ECM production, as well. Our results demonstrate that high dietary salt intake is associated with increased dermal pro-inflammatory status. Interestingly, although inflammation induces the synthesis of ECM in most organs, the expression of ECM decreased in the inflamed skin of mice on high salt diet. Our data suggest that salt intake may alter the process of skin remodeling.

Fibroblasts play a central role in diseases associated with excessive deposition of extracellular matrix (ECM), including idiopathic pulmonary fibrosis. Investigation of different properties of fibroblasts, such as migration, proliferation, and collagen-rich ECM production is unavoidable both in basic research and in the development of antifibrotic drugs. In the present study we developed a cost-effective, 96-well plate-based method to examine the migration of fibroblasts, as an alternative approach to the gold standard scratch assay, which has numerous limitations. This article presents a detailed description of our transient agarose spot (TAS) assay, with instructions for its routine application. Advantages of combined use of different functional assays for fibroblast activation in drug development are also discussed by examining the effect of nintedanib—an FDA approved drug against IPF—on lung fibroblasts.

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BACKGROUND AND AIMS Patients with chronic kidney disease (CKD) suffer from increased oxidative stress, which is further aggravated in patients on peritoneal dialysis (PD). Parkinson disease protein 7 (PARK7) has antioxidant and antiapoptotic activity; its role in PD is unknown. METHOD Transcriptome and proteome data sets from microdissected omental arterioles obtained from age-matched non-CKD children, children with CKD5 and children on PD with fluids containing low or high concentrations of glucose degradation products (GDP; n = 6/group) underwent PARK7 related gene set analysis (FDR < 0.05). Respective parietal peritoneal tissues (n = 60) underwent digital histomorphometry analyses. PARK7 western blotting was performed in effluents of eight children on high-GDP PD. Human umbilical endothelial cells (HUVEC) viability (MTT assay) and transepithelial electrical resistance (TER, Transwells) and 4-, 10- and 70-kDa dextran transport were measured (n = 6–12/group). As an extreme phenotype approach of PD toxicity, C57/BL6J mice were treated with chlorhexidine digluconate (CG) and PARK7 activator for 7 days and parietal peritoneal damage was quantitated (n = 6–8/group). RESULTS Arteriolar transcriptome analyses in children on low-GDP PD demonstrated the enrichment of PARK7 related GO terms of oxidant detoxification as compared to CKD5 and in children on high-GDP PD that of reactive oxygen species-, mitochondria- and apoptosis-related processes. On the proteome level the DNA repair/organization, catabolic and mitochondria associated processes were enriched in children on low-GDP PD, and mitochondrial processes in children on high-GDP PD. PARK7 was detected in the parietal peritoneal tissues in mesothelial, endothelial and inflammatory cells, in myocytes and fibroblasts and was present in the PD effluents. Total peritoneal and submesothelial PARK7 abundance was similar in controls, patients with CKD5 and in patients on low-GDP PD, but 2-fold increased in patients on high GDP PD compared to controls and CKD5. Mesothelial PARK7 was 2-fold increased in children on low-GDP PD versus CKD5, endothelial PARK7 abundance was similar in all four groups. In low-GDP PD patients endothelial PARK7 abundance correlated with vessel lumen/vessel diameter ratio (r = 0.53, P = 0.06), i.e. inversely with lumen obliteration. Submesothelial PARK7 correlated with microvessel density (r = 0.55, P = 0.05), with submesothelial hypoxia inducible factor-1 and angiopoietin-1 and -2 (ρ = 0.63, P = 0.023; r = 0.91, P < 0.0001; r = 0.60, P = 0.03) but not with VEGF. Submesothelial and endothelial PARK7 correlated with respective caspase 3 abundances (r = 0.74/0.68, P = 0.009/0.015). In HUVEC methylglyoxal (MG) dose- and time-dependently reduced viability, coincubation with PARK7 activator partially preserved endothelial cell viability. In Transwells, MG treatment decreased TER and increased dextran transport, but none of them was improved by PARK7 activation. In mice treated with CG submesothelial thickness was 2-fold increased, microvessel density was unchanged; PARK7 protein abundance was 5-fold reduced. Co-treatment of CG with PARK7 activator prevented the submesothelial thickening. CONCLUSION PD modifies arteriolar PARK7 related biological processes of oxidant detoxification, mitochondria- and apoptosis-related processes. PARK7 is ubiquitously expressed in the parietal peritoneum and regulated by the GDP content of PD fluids. In patients on low-GDP PD, PARK7 abundance correlated with the degree of arteriolar lumen narrowing, and VEGF-independent angiogenesis. Activation of PARK7 preserves endothelial cell viability in vitro and prevents CG induced peritoneal membrane damage in mice and thus represents a potential novel therapeutic approach.