Original

The Journal of the Korean Rheumatism Association 2003; 10(4): 402-412

Published online December 30, 2003

© Korean College of Rheumatology

류마티스관절 활막세포에서의 Receptor Activator of NF-κB Ligand (RANKL)의 발현 및 파골세포 생성

주영실·정명아·신동혁·임미경·심승철·정덕환*

을지대학교 의과대학 내과학교실, 경희대학교 의과대학 정형외과학교실*

Expression of Receptor Activator of NF-κB Ligand (RANKL) and Formation of Osteoclast in Cultured Synovial Fibroblasts

Yeong-Shil Joo, Myeong-A Cheong, Dong-Hyuk Sheen, Mi-Kyoung Lim, Seung Cheol Shim, Duke-Whan Chung*

Department of Internal Medicine, Eulji University School of Medicine and Department of Orthopedic Surgery, Kyung-Hee University College of Medicine*, Seoul, Korea

Correspondence to : Yeong-Shil Joo

Abstract

Objective: A number of soluble factors,which play important role in the pathophysiology of rheumatoid synovitis,are also known to be involved in osteoclast differentiation and activation through RANKL (Receptor activator of NF-κB ligand). To investigate the importance of RANKL in the pathogenesis of bone erosion in rheumatoid arthritis (RA) patients, we analyzed the expression of RANKL and Osteoprotegerin (OPG),and examined the formation of osteoclasts in rheumatoid synovial fibroblasts under the influence of various osteotropic factors.

Methods: Primary culture synoviocytes or fibroblast-like synoviocytes isolated from synovial tissues of 8 RA patients were cultured and treated with IL-1β (2 ng/ml), TNF-α (2 ng/ml), INF-γ(1000μ/ml), IL-15 (10 ng/ml), IL-12 (10 ng/ml), dexamethasone (10-9 M), PMA (10 ng/ml) or 1,25 (OH)2D3 (10-9 M) for 18 hours. Expression RANKL or OPG mRNA was measured by semiquantitative RT-PCR within linear amplification condition. TRAP (⁢) MNC (tartrate resistant acid phosphatase-positive multinucleated cell) formation was induced from primary culture synoviocytes or in coculture system of synovial fibroblasts with PBMCs in the presence of M-CSF and 1,25 (OH)2D3.

Results: 1. The intensity of base-line expression was different from patient to patient. Primary culture synoviocytes and synovial fibroblasts express RANKL and OPG mRNA with decreasing intensity when they are passaged. 2. Expresssion of RANKL mRNA was significantly increased by 1,25 (OH)2D3 and IL-1β (158.8⁑21% and 197.2⁑17% of controls, p<0.05 and p<0.005, respectively), while decreased significantly by dexamethasone (25.6⁑4.6% of controls, p<0.005). Expression of RANKL mRNA was significantly increased by IL-1β and decreased by dexamethasone, in a dose- and time-dependant manner. 3. TRAP (⁢) MNCs are formed from primary culture synoviocytes or in coculture system of synovial fibroblasts and PBMC in the presence of M-CSF and 1,25 (OH)2D3. Dexamethasone clearly inhibited TRAP (⁢) MNCs formation from synovial cells.

Conclusion: The regulatory mechanism for the expression of RANKL or OPG in rheumatoid synoviocytes might be different from that in bone marrow cells. Modulating the expression of these molecules could have potential therapeutic implication targeting bone destruction in RA.

Keywords RANKL, Rheumatoid arthritis, Fibroblast-like synoviocytes, Dexamethasone

Article

Original

The Journal of the Korean Rheumatism Association 2003; 10(4): 402-412

Published online December 30, 2003

Copyright © Korean College of Rheumatology.

류마티스관절 활막세포에서의 Receptor Activator of NF-κB Ligand (RANKL)의 발현 및 파골세포 생성

주영실·정명아·신동혁·임미경·심승철·정덕환*

을지대학교 의과대학 내과학교실, 경희대학교 의과대학 정형외과학교실*

Expression of Receptor Activator of NF-κB Ligand (RANKL) and Formation of Osteoclast in Cultured Synovial Fibroblasts

Yeong-Shil Joo, Myeong-A Cheong, Dong-Hyuk Sheen, Mi-Kyoung Lim, Seung Cheol Shim, Duke-Whan Chung*

Department of Internal Medicine, Eulji University School of Medicine and Department of Orthopedic Surgery, Kyung-Hee University College of Medicine*, Seoul, Korea

Correspondence to:Yeong-Shil Joo

Abstract

Objective: A number of soluble factors,which play important role in the pathophysiology of rheumatoid synovitis,are also known to be involved in osteoclast differentiation and activation through RANKL (Receptor activator of NF-κB ligand). To investigate the importance of RANKL in the pathogenesis of bone erosion in rheumatoid arthritis (RA) patients, we analyzed the expression of RANKL and Osteoprotegerin (OPG),and examined the formation of osteoclasts in rheumatoid synovial fibroblasts under the influence of various osteotropic factors.

Methods: Primary culture synoviocytes or fibroblast-like synoviocytes isolated from synovial tissues of 8 RA patients were cultured and treated with IL-1β (2 ng/ml), TNF-α (2 ng/ml), INF-γ(1000μ/ml), IL-15 (10 ng/ml), IL-12 (10 ng/ml), dexamethasone (10-9 M), PMA (10 ng/ml) or 1,25 (OH)2D3 (10-9 M) for 18 hours. Expression RANKL or OPG mRNA was measured by semiquantitative RT-PCR within linear amplification condition. TRAP (⁢) MNC (tartrate resistant acid phosphatase-positive multinucleated cell) formation was induced from primary culture synoviocytes or in coculture system of synovial fibroblasts with PBMCs in the presence of M-CSF and 1,25 (OH)2D3.

Results: 1. The intensity of base-line expression was different from patient to patient. Primary culture synoviocytes and synovial fibroblasts express RANKL and OPG mRNA with decreasing intensity when they are passaged. 2. Expresssion of RANKL mRNA was significantly increased by 1,25 (OH)2D3 and IL-1β (158.8⁑21% and 197.2⁑17% of controls, p<0.05 and p<0.005, respectively), while decreased significantly by dexamethasone (25.6⁑4.6% of controls, p<0.005). Expression of RANKL mRNA was significantly increased by IL-1β and decreased by dexamethasone, in a dose- and time-dependant manner. 3. TRAP (⁢) MNCs are formed from primary culture synoviocytes or in coculture system of synovial fibroblasts and PBMC in the presence of M-CSF and 1,25 (OH)2D3. Dexamethasone clearly inhibited TRAP (⁢) MNCs formation from synovial cells.

Conclusion: The regulatory mechanism for the expression of RANKL or OPG in rheumatoid synoviocytes might be different from that in bone marrow cells. Modulating the expression of these molecules could have potential therapeutic implication targeting bone destruction in RA.

Keywords: RANKL, Rheumatoid arthritis, Fibroblast-like synoviocytes, Dexamethasone

JRD
Jan 01, 2025 Vol.32 No.1, pp. 1~7
COVER PICTURE
Cumulative growth of rheumatology members and specialists (1980~2024). Cumulative distribution of the number of the (A) Korean College of Rheumatology members and (B) rheumatology specialists. (J Rheum Dis 2025;32:63-65)

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