ROCK2 regulates TGF-β-induced expression of CTGF and profibrotic genes via NF-κB and cytoskeleton dynamics in mesangial cells | JoVE Visualize

ROCK2 regulates TGF-β-induced expression of CTGF and profibrotic genes via NF-κB and cytoskeleton dynamics in mesangial cells

Yosuke Nagai¹, Keiichiro Matoba¹, Daiji Kawanami²

¹Division of Diabetes, Metabolism, and Endocrinology, Department of Internal Medicine, The Jikei University School of Medicine, Tokyo, Japan.

American Journal of Physiology. Renal Physiology|August 1, 2019

View abstract on PubMed

Summary

Rho kinase 2 (ROCK2) drives diabetic kidney disease progression by increasing profibrotic factors. Inhibiting ROCK2 reduces kidney damage and albuminuria in diabetic mice, highlighting ROCK2 as a therapeutic target.

Area of Science:

Nephrology
Molecular Biology
Pharmacology

Background:

Diabetic kidney disease (DKD) pathogenesis involves Rho kinase (ROCK).
ROCK has two isoforms, ROCK1 and ROCK2, with unclear roles in DKD.
Identifying specific ROCK isoforms involved is crucial for targeted therapies.

Purpose of the Study:

To determine the specific role of ROCK isoforms in diabetic glomerulosclerosis.
To investigate the mechanisms by which ROCK influences profibrotic mediator expression.
To evaluate the therapeutic potential of ROCK2 inhibition in a mouse model of type 2 diabetes.

Main Methods:

Isolated glomeruli and mesangial cells from diabetic (db/db) and control (db/m) mice.
Gene expression analysis (TGF-β, profibrotic mediators).
ROCK inhibition and siRNA knockdown (ROCK1, ROCK2).
Western blotting for phosphorylated JNK, Erk, and NF-κB.
In vivo studies with ROCK2 inhibitor administration.

Main Results:

Diabetic glomeruli showed increased TGF-β and profibrotic mediators.
ROCK inhibition suppressed profibrotic mediators via JNK, Erk, and NF-κB.
ROCK2 siRNA, but not ROCK1, inhibited fibrotic gene expression.
Elevated ROCK2 activity in diabetic mouse kidneys.
ROCK2 inhibitor treatment reduced albuminuria and glomerular fibrosis.

Conclusions:

ROCK2 is a key mediator of fibrotic processes in diabetic kidney disease.
ROCK2 activity drives the expression of profibrotic factors in glomeruli.
Targeting ROCK2 offers a promising therapeutic strategy for DKD.