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Effects of Tumor Necrosis Factor-α on Podocyte Expression of Monocyte Chemoattractant Protein-1 and in Diabetic
Choon Hee Chung1, Jingyi Fan2, Eun Young Lee3
1Division of Nephrology/Hypertension, Northwestern University, Chicago, Ill., USA ; Department of Internal Medicine, Yonsei University Wonju College of Medicine, Wonju, Korea.
Background/Aims:
Tumor necrosis factor (TNF)-α is believed to play a role in diabetic kidney disease. This study explores the specific effects of TNF-α with regard to nephropathy-relevant parameters in the podocyte.
Methods:
Cultured mouse podocytes were treated with recombinant TNF-α and assayed for production of monocyte chemoattractant protein-1 (MCP-1) by enzyme-linked immunosorbent assay (ELISA). TNF-α signaling of MCP-1 was elucidated by antibodies against TNF receptor (TNFR) 1 or TNFR2 or inhibitors of nuclear factor-kappaB (NF-κB), phosphatidylinositol 3-kinase (PI3K) or Akt. In vivo studies were done on male db/m and type 2 diabetic db/db mice. Levels of TNF-α and MCP-1 were measured by RT-qPCR and ELISA in the urine, kidney and plasma of the two cohorts and correlated with albuminuria.
Results:
Podocytes treated with TNF-α showed a robust increase (∼900%) in the secretion of MCP-1, induced in a dose- and time-dependent manner. Signaling of MCP-1 expression occurred through TNFR2, which was inducible by TNF-α ligand, but did not depend on TNFR1. TNF-α then proceeded via the NF-κB and the PI3K/Akt systems, based on the effectiveness of the inhibitors of those pathways. For in vivo relevance to diabetic kidney disease, TNF-α and MCP-1 levels were found to be elevated in the urine of db/db mice but not in the plasma.
Conclusion:
TNF-α potently stimulates podocytes to produce MCP-1, utilizing the TNFR2 receptor and the NF-κB and PI3K/Akt pathways. Both TNF-α and MCP-1 levels were increased in the urine of diabetic db/db mice, correlating with the severity of diabetic albuminuria.
Insights
Tumor necrosis factor-alpha (TNF-α) significantly increases monocyte chemoattractant protein-1 (MCP-1) in podocytes via TNFR2, NF-κB, and PI3K/Akt pathways. Elevated urinary TNF-α and MCP-1 correlate with diabetic kidney disease severity.
Area of Science:
- Nephrology
- Immunology
- Molecular Biology
Background:
- Diabetic kidney disease (DKD) is a major complication of diabetes.
- Tumor necrosis factor-alpha (TNF-α) is implicated in DKD pathogenesis.
- Podocyte dysfunction is central to DKD progression.
Purpose of the Study:
- To investigate the role of TNF-α in podocyte injury.
- To elucidate the signaling pathways involved in TNF-α-induced podocyte responses.
- To correlate TNF-α and MCP-1 levels with DKD in vivo.
Main Methods:
- Cultured mouse podocytes were treated with TNF-α.
- Monocyte chemoattractant protein-1 (MCP-1) production was measured by ELISA.
- Signaling pathways (TNFR1, TNFR2, NF-κB, PI3K/Akt) were analyzed using antibodies and inhibitors.
- In vivo studies used db/m and db/db mice, measuring urinary and plasma TNF-α and MCP-1.
Main Results:
- TNF-α treatment robustly increased MCP-1 secretion from podocytes in a dose- and time-dependent manner.
- TNF-α signaling occurred via TNFR2, NF-κB, and PI3K/Akt pathways, not TNFR1.
- Elevated urinary levels of TNF-α and MCP-1 were observed in diabetic db/db mice compared to controls.
Conclusions:
- TNF-α potently stimulates podocytes to produce MCP-1 through TNFR2, NF-κB, and PI3K/Akt.
- Increased urinary TNF-α and MCP-1 in db/db mice suggest their involvement in DKD.
- These findings highlight TNF-α as a potential therapeutic target in diabetic kidney disease.
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