Korean J Intern Med.  2019 Jan;34(1):210-219. 10.3904/kjim.2016.329.

N-acetyl-l-cysteine controls osteoclastogenesis through regulating Th17 differentiation and RANKL in rheumatoid arthritis

• Kim HR
• Kim KW
• Kim BM
• Lee KA
• Lee SH

• KMID: 2452505
• DOI: http://doi.org/10.3904/kjim.2016.329

Abstract

BACKGROUND/AIMS
This study aimed to determine the regulatory role of N-acetyl-l-cysteine (NAC), an antioxidant, in interleukin 17 (IL-17)-induced osteoclast differentiation in rheumatoid arthritis (RA).
METHODS
After RA synovial fibroblasts were stimulated by IL-17, the expression and production of receptor activator of nuclear factor κ-B ligand (RANKL) was determined by real-time polymerase chain reaction and enzyme-linked immunosorbent assay (ELISA). Osteoclastogenesis was also determined after co-cultures of IL-17-stimulated RA synovial fibroblasts, Th17 cells and various concentrations of NAC with monocytes. After human peripheral CD4⁺ T cells were cultured with NAC under Th17 condition, IL-17, interferon γ, IL-4, Foxp3, RANKL, and IL-2 expression and production was determined by flow cytometry or ELISA.
RESULTS
When RA synovial fibroblasts were stimulated by IL-17, IL-17 stimulated the production of RANKL, and NAC reduced the IL-17-induced RANKL production in a dose-dependent manner. NAC decreased IL-17-activated phosphorylation of mammalian target of rapamycin, c-Jun N-terminal kinase, and inhibitor of κB. When human peripheral blood CD14⁺ monocytes were cultured with macrophage colony-stimulating factor and IL-17 or RANKL, osteoclasts were differentiated, and NAC reduced the osteoclastogenesis. After human peripheral CD4⁺ T cells were co-cultured with IL-17-pretreated RA synovial fibroblasts or Th17 cells, NAC reduced their osteoclastogenesis. Under Th17 polarizing condition, NAC decreased Th17 cell differentiation and IL-17 and RANKL production.
CONCLUSIONS
NAC inhibits the IL-17-induced RANKL production in RA synovial fibroblasts and IL-17-induced osteoclast differentiation. NAC also reduced Th17 polarization. NAC could be a supplementary therapeutic option for inflammatory and bony destructive processes in RA.

Keyword

Acetylcysteine; Arthritis, rheumatoid; Osteogenesis; RANK ligand; Interleukin-17

MeSH Terms

Acetylcysteine*
Arthritis, Rheumatoid*
Coculture Techniques
Enzyme-Linked Immunosorbent Assay
Fibroblasts
Flow Cytometry
Humans
Interferons
Interleukin-17
Interleukin-2
Interleukin-4
JNK Mitogen-Activated Protein Kinases
Macrophage Colony-Stimulating Factor
Monocytes
Osteoclasts
Osteogenesis
Phosphorylation
RANK Ligand
Real-Time Polymerase Chain Reaction
Sirolimus
T-Lymphocytes
Th17 Cells
Acetylcysteine
Interferons
Interleukin-17
Interleukin-2
Interleukin-4
JNK Mitogen-Activated Protein Kinases
Macrophage Colony-Stimulating Factor
RANK Ligand
Sirolimus