Novel inhibitors of the cellular renin-angiotensin system components, poricoic acids, target Smad3 phosphorylation and Wnt/β-catenin pathway against renal fibrosis

Br J Pharmacol. 2018 Jul;175(13):2689-2708. doi: 10.1111/bph.14333. Epub 2018 May 22.

Abstract

Background and purpose: Tubulo-interstitial fibrosis is the final pathway in the progression of chronic kidney disease (CKD) to kidney failure. The renin-angiotensin system (RAS) plays a major role in CKD progression. Hence, we determined the efficacy of novel RAS inhibitors isolated from Poria cocos against renal fibrosis.

Experimental approach: Effects of three novel tetracyclic triterpenoid compounds, poricoic acid ZC (PZC), poricoic acid ZD (PZD) and poricoic acid ZE (PZE), were investigated on TGFβ1- and angiotensin II (AngII)-treated HK-2 cells and unilateral ureteral obstruction (UUO) in mice. Immunofluorescence staining, quantitative real-time PCR, siRNA, co-immunoprecipitation and Western blot analyses were used to evaluate expression of key molecules in RAS, Wnt/β-catenin and TGFβ/Smad pathways.

Key results: Addition of the above compounds to culture media and their administration to UUO mice: (i) significantly attenuated epithelial-to-mesenchymal transition and extracellular matrix production in TGFβ1- and AngII-treated HK-2 cells and UUO mice by inhibiting Wnt/β-catenin pathway activation and Smad3 phosphorylation; (ii) selectively inhibited Smad3 phosphorylation by blocking the interaction of TGFBR1 with Smad3; and (iii) specifically inhibited Smad3 activation. PZC and PZD showed a strong inhibitory effect on all RAS components, and PZE showed a strong inhibitory effect on renin. Furthermore, the secolanostane tetracyclic triterpenoids, PZC and PZD, showed a stronger inhibitory effect than the lanostane tetracyclic triterpenoid PZE. Therefore, compounds with secolanostance skeleton showed stronger bioactivity than those with lanostance skeleton.

Conclusion and implications: The secolanostane tetracyclic triterpenoids effectively blocked RAS by simultaneously targeting multiple RAS components and lanostane tetracyclic triterpenoids inhibited renin and protected against tubulo-interstitial fibrosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Survival / drug effects
  • Cells, Cultured
  • Fibrosis / drug therapy*
  • Fibrosis / pathology
  • Humans
  • Kidney / drug effects*
  • Kidney / pathology
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Molecular Conformation
  • Phosphorylation / drug effects
  • Renin-Angiotensin System / drug effects*
  • Smad3 Protein / antagonists & inhibitors*
  • Smad3 Protein / metabolism
  • Triterpenes / chemistry
  • Triterpenes / pharmacology*
  • Wnt Signaling Pathway / drug effects*
  • beta Catenin / antagonists & inhibitors*
  • beta Catenin / metabolism

Substances

  • SMAD3 protein, human
  • Smad3 Protein
  • Smad3 protein, mouse
  • Triterpenes
  • beta Catenin
  • poricoic acid A