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Updated: Mar 22, 2026
Molecular Analysis of Endothelial-mesenchymal Transition Induced by Transforming Growth Factor-β Signaling
Published on: August 3, 2018
Matrix metalloproteinase 9 induces endothelial-mesenchymal transition via Notch activation in human kidney glomerular
Ye Zhao1,2, Xi Qiao1,3, Lihua Wang3
1Centre for Transplant and Renal Research, Westmead Institute for Medical Research, the University of Sydney, 176 Hawkesbury Road, Sydney, NSW, 2145, Australia.
Background:
Endothelial-mesenchymal transition (EndoMT) is a major source of myofibroblast formation in kidney fibrosis. Our previous study showed a profibrotic role for matrix metalloproteinase 9 (MMP-9) in kidney fibrosis via induction of epithelial-mesenchymal transition (EMT). Inhibition of MMP-9 activity reduced kidney fibrosis in murine unilateral ureteral obstruction. This study investigated whether MMP-9 also plays a role in EndoMT in human glomerular endothelial cells.
Results:
TGF-β1 (10 or 20 ng/ml) induced EndoMT in HKGECs as shown by morphological changes. In addition, VE-cadherin and CD31 were significantly downregulated, whereas α-SMA, vimentin, and N-cadherin were upregulated. RT-PCR revealed that Snail, a known inducer of EMT, was upregulated. The MMP inhibitor GM6001 abrogated TGF-β1-induced EndoMT. Zymography indicated that MMP-9 was also upregulated in TGF-β1-treated HKGECs. Recombinant MMP-9 (2 μg/ml) induced EndoMT in HKGECs via Notch signaling, as evidenced by increased formation of the Notch intracellular domain (NICD) and decreased Notch 1. Inhibition of MMP-9 activity by its inhibitor showed a dose-dependent response in preventing TGF-β1-induced α-SMA and NICD in HKGECs, whereas inhibition of Notch signaling by γ-secretase inhibitor (GSI) blocked rMMP-9-induced EndoMT.
Conclusions:
Taken together, our results demonstrate that MMP-9 plays an important role in TGF-β1-induced EndoMT via upregulation of Notch signaling in HKGECs.
Insights
Matrix metalloproteinase 9 (MMP-9) drives kidney fibrosis by promoting endothelial-mesenchymal transition (EndoMT) through Notch signaling. Inhibiting MMP-9 activity effectively prevents EndoMT in human glomerular endothelial cells.
Area of Science:
- Nephrology
- Cell Biology
- Molecular Biology
Background:
- Endothelial-mesenchymal transition (EndoMT) is a key process in kidney fibrosis.
- Matrix metalloproteinase 9 (MMP-9) has a profibrotic role, previously linked to epithelial-mesenchymal transition (EMT).
- This study investigates MMP-9's role in EndoMT within human glomerular endothelial cells.
Purpose of the Study:
- To determine if MMP-9 contributes to TGF-β1-induced EndoMT in human glomerular endothelial cells.
- To elucidate the signaling pathways involved in MMP-9-mediated EndoMT.
Main Methods:
- Human glomerular endothelial cells (HKGECs) were treated with TGF-β1.
- Morphological changes and protein expression (VE-cadherin, CD31, α-SMA, vimentin, N-cadherin) were assessed.
- RT-PCR, zymography, and specific inhibitors (GM6001, γ-secretase inhibitor) were used to evaluate MMP-9 and Notch signaling involvement.
Main Results:
- TGF-β1 induced EndoMT in HKGECs, characterized by altered cell morphology and marker expression.
- MMP-9 and Snail were upregulated following TGF-β1 treatment.
- Recombinant MMP-9 induced EndoMT via Notch signaling (increased NICD), and MMP-9 inhibition dose-dependently blocked TGF-β1-induced EndoMT and NICD.
Conclusions:
- MMP-9 plays a significant role in TGF-β1-induced EndoMT in human glomerular endothelial cells.
- The mechanism involves the upregulation of Notch signaling.
- Targeting MMP-9 presents a potential therapeutic strategy for kidney fibrosis.
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Epithelial-to-Mesenchymal Transition
The epithelial-to-mesenchymal transition or EMT is a developmental process commonly observed in wound healing, embryogenesis, and cancer metastasis. EMT is induced by transforming growth factor-beta (TGF-β) or receptor tyrosine kinase (RTK) ligands, which further...