S100A9 promotes renal calcium oxalate stone formation via activating the TLR4-p38/MAPK-LCN2 signaling pathway

Int J Biol Macromol. 2024 Nov;281(Pt 1):136178. doi: 10.1016/j.ijbiomac.2024.136178. Epub 2024 Sep 30.

Abstract

Objectives: To explore the role of S100A9 protein in renal calcium oxalate (CaOx) stone formation.

Methods: CaOx nephrocalcinosis mice were established via intraperitoneal injection of glyoxylate. They were treated with S100A9 deficiency, Paquinimod, or p38 MAPK-IN-1. Vonkossa staining was conducted to observe the deposition of CaOx crystals. Renal expression of inflammation, macrophage polarization, and injury markers was detected using immunohistochemistry and qPCR. Effects of S100A9 on renal tubular epithelial cells (HK-2) were explored by transcriptome sequencing. The mechanism of how S100A9 regulated lipocalin 2 (LCN2) was studied through Western Blot. Flow cytometry was performed to detect the influence of LCN2 on macrophages polarization.

Results: S100A9 deficiency inhibited the renal deposition of CaOx crystals in nephrocalcinosis mice. S100A9 upregulated the expression of LCN2 in HK-2 cells via activating the TLR4-p38/MAPK pathway. LCN2 promoted the migration and M1 polarization of macrophages. S100A9 deficiency downregulated the renal expression of LCN2, IL1-β, Kim-1, F4/80, and CD80 in nephrocalcinosis mice. Paquinimod and p38 MAPK-IN-1 both inhibited the renal deposition of CaOx crystals and downregulated the expression of LCN2, IL1-β, Kim-1, F4/80, iNOS, and CD68 in nephrocalcinosis mice.

Conclusions: S100A9 promotes renal inflammatory injury by activating the TLR4-p38/MAPK-LCN2 pathway and then contributes to CaOx stone formation.

Keywords: Inflammation; Renal calcium oxalate stones; S100A9.

MeSH terms

  • Animals
  • Calcium Oxalate* / metabolism
  • Calgranulin B* / genetics
  • Calgranulin B* / metabolism
  • Cell Line
  • Disease Models, Animal
  • Humans
  • Kidney / metabolism
  • Kidney / pathology
  • Kidney Calculi / etiology
  • Kidney Calculi / metabolism
  • Kidney Calculi / pathology
  • Lipocalin-2* / genetics
  • Lipocalin-2* / metabolism
  • MAP Kinase Signaling System / drug effects
  • Macrophages / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Signal Transduction
  • Toll-Like Receptor 4* / genetics
  • Toll-Like Receptor 4* / metabolism
  • p38 Mitogen-Activated Protein Kinases* / metabolism

Substances

  • Toll-Like Receptor 4
  • Lipocalin-2
  • Calgranulin B
  • Calcium Oxalate
  • p38 Mitogen-Activated Protein Kinases
  • Lcn2 protein, mouse
  • S100A9 protein, mouse
  • Tlr4 protein, mouse