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Induction and Analysis of Epithelial to Mesenchymal Transition

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Published on: August 27, 2013

Gremlin-mediated decrease in bone morphogenetic protein signaling promotes aristolochic acid-induced

Yi Li¹, Zihua Wang, Shuai Wang

¹Institute of Nephrology of Chongqing and Department of Nephrology, Xinqiao Hospital, Third Military Medical University, Chongqing, China.

|April 25, 2012

Summary

Aristolochic acid (AA) causes kidney damage by promoting epithelial-to-mesenchymal transition (EMT). Gremlin, a BMP-7 antagonist, exacerbates AA-induced EMT in kidney cells. Inhibiting gremlin may offer new treatments for aristolochic acid nephropathy (AAN).

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Area of Science:

Nephrology
Cell Biology
Molecular Medicine

Background:

Aristolochic acid (AA) ingestion causes aristolochic acid nephropathy (AAN), leading to kidney fibrosis, failure, and urothelial cancer.
Bone morphogenetic protein-7 (BMP-7) has shown potential in mitigating AA-induced epithelial-to-mesenchymal transition (EMT) in kidney cells.
The antagonistic role of gremlin in BMP-7's protective action against AA-induced kidney damage was previously unclear.

Purpose of the Study:

To investigate the specific role of gremlin in AA-induced EMT in human proximal tubule epithelial cells (PTEC).
To elucidate the mechanism by which gremlin antagonizes BMP-7 signaling in the context of AA exposure.

Main Methods:

Human proximal tubule epithelial cells (HK-2) were treated with aristolochic acid (AA).
Assessed cell viability (MTT assay), morphology (microscopy), and EMT markers (E-cadherin, α-SMA) via immunofluorescence.
Analyzed BMP-7 and gremlin expression (mRNA/protein) and BMP-7 activity (phosphorylated Smad1/5/8) using RT-PCR and Western blotting.
Utilized gremlin siRNA to evaluate the effect of gremlin knockdown on AA-induced EMT.

Main Results:

AA exposure induced EMT in HK-2 cells, characterized by altered cell shape, reduced E-cadherin, and increased α-SMA and collagen type I.
AA significantly increased gremlin expression while decreasing BMP-7 expression and BMP-7 signaling activity (p-Smad1/5/8).
Gremlin knockdown using siRNA restored BMP-7 signaling and attenuated AA-induced EMT phenotypes.

Conclusions:

Gremlin plays a crucial role in mediating the detrimental effects of AA on kidney proximal tubule cells.
Increased gremlin expression contributes to the loss of BMP-7's reno-protective function in AA-induced nephropathy.
Targeting gremlin presents a promising therapeutic strategy for developing novel treatments for AAN.