Carvacrol ameliorates skin allograft rejection through modulating macrophage polarization by activating the Wnt signalling pathway

Phytother Res. 2024 Sep;38(9):4675-4694. doi: 10.1002/ptr.8282. Epub 2024 Aug 9.

Abstract

Post-transplantation immune rejection remains an important factor for transplant patients. However, conventional immunosuppressants are associated with substantial adverse effects. Natural immunosuppressants present a promising alternative to conventional counterparts, boasting exceptional biological activity, minimal toxicity and reduced side effects. We identified carvacrol as a prospective immunosuppressive agent following T cell proliferation experiment and validated carvacrol's immunosuppressive efficacy in the murine allogeneic skin graft model. T cell proliferation assay was used to screen natural small molecule compounds and the immunosuppressive effect of compounds was evaluated in MHC-mismatched murine allogeneic skin graft model. H&E and immunohistochemical staining were applied to evaluate the pathological grade. Furthermore, flow cytometry was uitlized to analyse the immunophenotype changes of immune cells. Western blotting and q-PCR were used to detect the expression of key molecules in macrophages. In vitro, carvacrol demonstrates significant inhibition of the proliferation of CD4+ T and CD8+ T cells. It notably reduces inflammatory factor expression within the allografts, suppresses T cell differentiation toward Th1 phenotype and expansion. Furthermore, carvacrol prominently hinders M1-type macrophages polarization by activating Wnt signaling. Notably, the anti-rejection efficacy of carvacrol was significantly weakened upon the removal of macrophages in mice using chlorophosphate liposomes. Carvacrol could significantly inhibit T cell proliferation, alleviate graft rejection and has outstanding toxicological safety. The molecular mechanism of the anti-rejection effect of carvacrol is closely related to its mediating activation of macrophage Wnt pathway, inhibiting M1 polarization and inducing T cell differentiation.

Keywords: T cells; Wnt signalling pathway; carvacrol; inflammation; macrophage; skin allograft.

MeSH terms

  • Allografts
  • Animals
  • CD8-Positive T-Lymphocytes / drug effects
  • Cell Differentiation / drug effects
  • Cell Proliferation / drug effects
  • Cymenes* / pharmacology
  • Graft Rejection* / drug therapy
  • Graft Rejection* / prevention & control
  • Immunosuppressive Agents* / pharmacology
  • Macrophages* / drug effects
  • Male
  • Mice
  • Mice, Inbred BALB C*
  • Mice, Inbred C57BL*
  • Skin Transplantation* / adverse effects
  • T-Lymphocytes / drug effects
  • Wnt Signaling Pathway* / drug effects

Substances

  • carvacrol
  • Cymenes
  • Immunosuppressive Agents