Transcriptomic study of high-glucose effects on human skin fibroblast cells

Abstract

Abstract. Skin ulcers are a common complication of diabetes mellitus (DM). Fibroblasts are located within the dermis of skin tissue and can be damaged by diabetes. However, the underlying mechanism of how DM affects fibroblasts remains elusive. To understand the effects of DM on fibroblasts, the current study mimicked DM by high-glucose (HG) supplementation in the culture medium of human foreskin primary fibroblast cells, and the analysis of transcriptomic changes was conducted. RNA sequencing-based transcriptome analysis identified that, upon HG stress, 463 genes were upregulated and 351 genes downregulated (>1.5-fold changes; P<0.05). These altered genes were distributed into 20 different pathways. In addition, gene ontology (GO) analysis indicated that 31 GO terms were enriched. Among the pathways identified, nuclear factor κB (NF-κB) pathway genes were highly expressed, and the addition of Bay11-7082, a typical NF-κB signaling inhibitor, blocked the previously observed alterations in plasminogen activator inhibitor 1 (PAI1), an inflammation marker and frizzled class receptor 8 (FZD8), a Wnt signaling gene, expression that resulted from HG stress. Furthermore, an inhibitor of Wnt signaling diminished the role of Bay11-7082 in the regulation of PAI1 expression under HG conditions, suggesting that Wnt signaling may function downstream of the NF-κB pathway to protect fibroblast cells from HG stress. To the best of our knowledge, the current study is the first analysis of transcriptomic responses under HG stress in human fibroblasts. The data provided here may aid the understanding of the molecular mechanisms by which fibroblast cells are damaged in the skin of patients with DM.

Sonic hedgehog‑c‑Jun N‑terminal kinase‑zinc finger protein Gli1 signaling protects against high glucose concentration‑induced reactive oxygen species generation in human fibroblasts

“…Initially, HG concentration was optimized for the relevant experimental conditions prior to testing gene expression. Previous research has identified that HG concentrations >35 mM significantly inhibit cell proliferation (11). In the present study, it was confirmed that 35 mM HG stress significantly reduced fibroblast proliferation, in comparison with LG fibroblasts (Fig.…”

“…To understand the mechanism of diabetes, HG is typically used to mimic the disease. Previous studies have observed that HG mediated stress altered the expression of many genes including hedgehog signaling-related genes in human fibroblast primary cells (11). To analyze the effect of HG on hedgehog signaling, key genes associated the pathway were examined for HG-mediated changes in expression.…”

“…In addition, it has been demonstrated that cell proliferation was defective and subsequent production of extracellular matrix protein was affected in diabetic ulcer fibroblasts (8 3 Pathology and 4 Immunology, c-Jun N-terminal kinase (JNK) activity, resulting in a delay in fibroblast migration (10). Previous transcriptome analysis using HG-stimulated fibroblasts has identified changes in the expression of numerous genes associated with diverse pathways including Wnt, inflammatory response, and hedgehog signaling (11). Hedgehog signaling has been widely studied due to its significance in development and disease regulation (12).…”

Nuclear factor-κB signaling negatively regulates high glucose-induced vascular endothelial cell damage downstream of the extracellular signal-regulated kinase/c-Jun N-terminal kinase pathway

“…In addition, NOD-like receptor signaling was altered upon HG stress in VECs. Decreased NOD-like signaling in human fibroblast cells with HG stress were also observed in a previous study (12). Furthermore, genetic variation of NOD1 and NOD2 has been associated with type II DM (23), suggesting the importance of NOD-like signaling in DM-mediated stress.…”

“…2 according to their rankings, including alcoholism as well as NF-κB and extracellular matrix (ECM)-receptor interaction signaling pathways. NOD-like receptor signaling was identified in VEC cell with HG stress, which was previously identified as a target pathway in HG-stressed fibroblast cells (12). These data indicated that HG regulates a large number of genes involved in different biological processes.…”