Renoprotective effect of rosmarinic acid by inhibition of indoxyl sulfate-induced renal interstitial fibrosis via the NLRP3 inflammasome signaling

Int Immunopharmacol. 2024 Jun 30:135:112314. doi: 10.1016/j.intimp.2024.112314. Epub 2024 May 23.

Abstract

We previously reported that rosmarinic acid (RA) ameliorated renal fibrosis in a unilateral ureteral obstruction (UUO) murine model of chronic kidney disease. This study aimed to determine whether RA attenuates indoxyl sulfate (IS)-induced renal fibrosis by regulating the activation of the NLRP3 inflammasome/IL-1β/Smad circuit. We discovered the NLRP3 inflammasome was activated in the IS treatment group and downregulated in the RA-treated group in a dose-dependent manner. Additionally, the downstream effectors of the NLRP3 inflammasome, cleaved-caspase-1 and cleaved-IL-1β showed similar trends in different groups. Moreover, RA administration significantly decreased the ROS levels of reactive oxygen species in IS-treated cells. Our data showed that RA treatment significantly inhibited Smad-2/3 phosphorylation. Notably, the effects of RA on NLRP3 inflammasome/IL-1β/Smad and fibrosis signaling were reversed by the siRNA-mediated knockdown of NLRP3 or caspase-1 in NRK-52E cells. In vivo, we demonstrated that expression levels of NLRP3, c-caspase-1, c-IL-1β, collagen I, fibronectin and α-SMA, and TGF- β 1 were downregulated after treatment of UUO mice with RA or RA + MCC950. Our findings suggested RA and MCC950 synergistically inhibited UUO-induced NLRP3 signaling activation, revealing their renoprotective properties and the potential for combinatory treatment of renal fibrosis and chronic kidney inflammation.

Keywords: Indoxyl sulfate; MCC950; NLRP3 inflammasome; Rosmarinic acid; Unilateral ureteral obstruction.

MeSH terms

  • Animals
  • Caspase 1 / metabolism
  • Cell Line
  • Cinnamates* / pharmacology
  • Cinnamates* / therapeutic use
  • Depsides* / pharmacology
  • Depsides* / therapeutic use
  • Disease Models, Animal
  • Fibrosis*
  • Indican*
  • Inflammasomes* / metabolism
  • Interleukin-1beta / metabolism
  • Kidney Diseases / chemically induced
  • Kidney Diseases / drug therapy
  • Kidney Diseases / pathology
  • Kidney* / drug effects
  • Kidney* / metabolism
  • Kidney* / pathology
  • Male
  • Mice
  • Mice, Inbred C57BL*
  • NLR Family, Pyrin Domain-Containing 3 Protein* / metabolism
  • Reactive Oxygen Species / metabolism
  • Renal Insufficiency, Chronic / chemically induced
  • Renal Insufficiency, Chronic / drug therapy
  • Renal Insufficiency, Chronic / metabolism
  • Renal Insufficiency, Chronic / pathology
  • Rosmarinic Acid*
  • Signal Transduction* / drug effects
  • Smad2 Protein / metabolism
  • Smad3 Protein / metabolism
  • Ureteral Obstruction / drug therapy
  • Ureteral Obstruction / pathology

Substances

  • Rosmarinic Acid
  • Depsides
  • Cinnamates
  • NLR Family, Pyrin Domain-Containing 3 Protein
  • Inflammasomes
  • Indican
  • Nlrp3 protein, mouse
  • Interleukin-1beta
  • Reactive Oxygen Species
  • Smad2 Protein
  • Smad3 Protein
  • Caspase 1