The implication of Numb-induced Notch signaling in endothelial-mesenchymal transition of diabetic nephropathy

J Diabetes Complications. 2018 Oct;32(10):889-899. doi: 10.1016/j.jdiacomp.2018.06.011. Epub 2018 Jul 10.

Abstract

Aim: This study was purposed to figure out the contribution of Numb-induced Notch signaling to the development of diabetic nephropathy (DN).

Methods: Two hundred and twenty six DN patients were included, and human glomerular endothelial cells (RGEC) were cultured. MSCV-Numb-IRES-GFP, MSCV-Notch1-IRES-GFP and MSCV-Hes1-IRES-GFP were transfected to construct the recombinant retroviral vectors.

Result: The over-expressed Numb and Notch1, as well as the under-expressed Hes-1 were correlated with the undesirable prognosis of DN patients (P < 0.05). Within the cell lines transfection with si-Numb would cut down E-cadherin and CD31 expressions (P < 0.05), yet elevated α-SMA and vimentin expressions (P < 0.05). The apoptotic rate of si-Numb cell lines underperformed ones categorized into the hyperglucose group (P < 0.05), whereas the lowly-expressed Notch1 and Hes1 were observably associated with inhibited proliferation of myofibroblasts (P < 0.05). Addition of ADPT caused under-expressed α-SMA and vimentin, along with the over-expressed E-cadherin and CD31 (P < 0.05). Silencing of Notch1 and Hes1 reversed the epithelial-mesenchymal transition (EMT) process that was triggered by high glucose (P < 0.05).

Conclusion: Numb negatively regulated Notch signaling pathway in EMT of DN, implying that they had great potentials to serve as therapeutic targets or diagnostic biomarkers for DN.

Keywords: Diabetic nephropathy; Endothelial-mesenchymal transition; Hes1; Notch signaling; Numb.

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Cells, Cultured
  • Diabetic Nephropathies / diagnosis
  • Diabetic Nephropathies / genetics*
  • Diabetic Nephropathies / metabolism
  • Diabetic Nephropathies / mortality
  • Epithelial-Mesenchymal Transition / genetics*
  • Female
  • Gene Expression
  • Glucose / pharmacology
  • HEK293 Cells
  • Humans
  • Male
  • Membrane Proteins / genetics
  • Membrane Proteins / physiology*
  • Middle Aged
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / physiology*
  • Receptor, Notch1 / genetics
  • Receptor, Notch1 / metabolism
  • Receptors, Notch / genetics*
  • Receptors, Notch / metabolism
  • Signal Transduction / drug effects
  • Signal Transduction / genetics
  • Signal Transduction / physiology
  • Transcription Factor HES-1 / genetics
  • Transcription Factor HES-1 / metabolism

Substances

  • Membrane Proteins
  • Nerve Tissue Proteins
  • NUMB protein, human
  • Receptor, Notch1
  • Receptors, Notch
  • Transcription Factor HES-1
  • HES1 protein, human
  • Glucose