Inhibition of ROCK2 alleviates renal fibrosis and the metabolic disorders in the proximal tubular epithelial cells.

Abstract

Non-specific inhibition of Rho-associated kinases (ROCKs) alleviated renal fibrosis in the unilateral ureteral obstruction (UUO) model, while genetic deletion of ROCK1 did not affect renal pathology in mice. Thus, whether ROCK2 plays a role in renal tubulointerstitial fibrosis needs to be clarified. In the present study, a selective inhibitor against ROCK2 or genetic approach was used to investigate the role of ROCK2 in renal tubulointerstitial fibrosis. In the fibrotic kidneys of chronic kidney diseases (CKDs) patients, we observed an enhanced expression of ROCK2 with a positive correlation with interstitial fibrosis. In mice, the ROCK2 protein level was time-dependently increased in the UUO model. By treating CKD animals with KD025 at the dosage of 50 mg/kg/day via intraperitoneal injection, the renal fibrosis shown by Masson's trichrome staining was significantly alleviated along with the reduced expression of fibrotic genes. In vitro, inhibiting ROCK2 by KD025 or ROCK2 knockdown/knockout significantly blunted the pro-fibrotic response in transforming growth factor-β1 (TGF-β1)-stimulated mouse renal proximal tubular epithelial cells (mPTCs). Moreover, impaired cellular metabolism was reported as a crucial pathogenic factor in CKD. By metabolomics analysis, we found that KD025 restored the metabolic disturbance, including the impaired glutathione metabolism in TGF-β1-stimulated tubular epithelial cells. Consistently, KD025 increased antioxidative stress enzymes and nuclear erythroid 2-related factor 2 (Nrf2) in fibrotic models. In addition, KD025 decreased the infiltration of macrophages and inflammatory response in fibrotic kidneys and blunted the activation of macrophages in vitro. In conclusion, inhibition of ROCK2 may serve as a potential novel therapy for renal tubulointerstitial fibrosis in CKD.

• Authors: Ran You, Wei Zhou, Yanwei Li, Yue Zhang, Songming Huang, Zhanjun Jia, Aihua Zhang
• Journal: Clinical science (London, England : 1979) (Clin Sci (Lond)) Vol. 134 Issue 12 Pg. 1357-1376 (06 26 2020) ISSN: 1470-8736 [Electronic] England
• PMID: 32490513 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2020 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Chemical References

• Anti-Inflammatory Agents
• Heterocyclic Compounds, 4 or More Rings
• KD025
• NF-E2-Related Factor 2
• Nfe2l2 protein, mouse
• Protein Kinase Inhibitors
• Smad2 Protein
• Transforming Growth Factor beta1
• ROCK2 protein, human
• Rock2 protein, mouse
• rho-Associated Kinases

Topics

• Adolescent
• Animals
• Anti-Inflammatory Agents (pharmacology)
• Child
• Child, Preschool
• Disease Models, Animal
• Epithelial Cells (drug effects, enzymology)
• Female
• Fibrosis
• Heterocyclic Compounds, 4 or More Rings (pharmacology)
• Humans
• Infant
• Inflammation (pathology)
• Kidney Tubules, Proximal (pathology)
• Macrophages (drug effects, metabolism)
• Male
• Metabolic Diseases (enzymology, pathology)
• Mice
• NF-E2-Related Factor 2 (metabolism)
• Oxidative Stress (drug effects)
• Protein Kinase Inhibitors (pharmacology)
• RAW 264.7 Cells
• Smad2 Protein (metabolism)
• Transforming Growth Factor beta1 (pharmacology)
• Up-Regulation (drug effects)
• Ureteral Obstruction (enzymology, pathology)
• rho-Associated Kinases (antagonists & inhibitors, metabolism)