• The Korean Journal of Pain
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• 제13권1호
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• pp.1-7
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• 2000

Background: Intraarticular local anesthetic injection has been therapeutically applied for pain control in various arthritis patients. However, little physiologic effects of local anesthetics on their tissue were known. This study was conducted to determine its effects on the cell proliferation and matrix metalloproteinases (MMP) production of cultured fibroblast like synoviocytes (FLS) derived from synovial tissues of rheumatoid arthritis patients. Methods: Bupivacaine with varying concentrations 0 (control), 0.1, 0.25, 0.5% was applied to experimental cell groups growing as monolayers in culture plates for varying durations 0 (control), 30, 90, 180 seconds in the presence and absence of interleukin-$1\beta$. Results: No statistical significances were noted in thymidine incorporation between 0, 30, 90 and 180 seconds exposure groups with 0.5% bupivacaine after 1 day and 2 days. Thymidine incorporation between 0, 0.1, 0.25, 0.5% exposure groups 1 day and 2 days after 90 seconds exposure did not show any differences. After exposure to bupivacaine, there were statistically significant increases in MMP-1 (p=0.025) and MMP-3 productions (p=0.000) of FLS in the absence of IL-$1\beta$, but no differences among the groups in the presence of IL-$1\beta$. Conclusion: We concluded that in this short-term in vitro study, bupivacaine does not have injurious effect on cultured rheumatoid arthritic joint tissues. The long-term effect cannot be known from this investigation.

• Molecular & Cellular Toxicology
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• 제3권2호
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• pp.85-89
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• 2007

Efficient gene delivery to specific tissues, such as inflammatory and cancerous tissues, is currently a major concern in disease treatment. The human transferrin receptor (hTR) has been detected in the synovium and fibroblast-like synoviocytes (FLS), which raises the possibility that expression of hTR is associated with the pathogenesis of rheumatoid arthritis (RA). To investigate whether the hTR is a useful target for gene transduction into the FLS of RA patients, recombinant adenoviruses with wildtype fiber (AdLac) and transferrin peptide-tagged fiber (Tf-AdLac) were used. The hTR expression level in FLS was notably increased by basic fibroblast growth factor (bFGF). Gene transduction to FLS was significantly higher by the hTR-targeted adenovirus than by the AdLac adenovirus, and treatment of the FLS with bFGF resulted in increased gene transduction by Tf-AdLac. Taken together, these data support Tf-AdLac as a new strategy for gene transduction in the treatment of RA patients.

• 동의생리병리학회지
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• 제19권6호
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• pp.1647-1655
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• 2005

Gamimahaenggamsuk-tang (GMHGST) is used for treatment of inflammatory diseases including rheumatoid arthritis (RA). Here, regulatory activity of GMHGST on RA-mediated inflammatory responses was investigated in cultured human fiDroblast-like synoviocytes (FLS), Levels of mRNAs encoding for inflammatory cytokines such as $IL-1{\beta}$, IL-6 and IL-8 and NOS-II enzyme, which had been induced by $TNF-{\alpha}$ and $IL-1{\beta}$ cotreatment, were decreased to the similar levels as those in cells treated with anti-inflammatory agent MTX. mRNA expressions of matrix metalloproteinase-3 (MMP-3) and tissue inhibitor of metalloproteinases (TIMPs) as well as intercellular adhesion molecule (ICAM) were also downregulated by increasing doses of GMHGST in activated FLS. Moreover, GMHGST appeared to protect cells by decreasing NO levels, and inhibited cell proliferation which had been induced by inflammatory stimulation by $TNF-{\alpha}$ and IL-1. These results suggest that GMHGST is effective as an inhibitory agent for regulating inflammatory responses in activated FLS.

• Molecules and Cells
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• 제39권8호
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• pp.611-618
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• 2016

Fibroblast-like synoviocytes (FLS) with aberrant expression of microRNA (miRNA) are critical pathogenic regulators in rheumatoid arthritis (RA). Previous studies have found that overexpression or silencing of miRNA can contribute to the development of miRNA-based therapeutics in arthritis models. In this study, we explored the effects of miR-27a on cell migration and invasion in cultured FLS from RA patients. We found that miR-27a was markedly downregulated in the serum, synovial tissue, and FLS of RA patients. Meanwhile, the expression of follistatin-like protein 1 (FSTL1) was upregulated, which suggests that FSTL1 plays a key role in RA development. The results of a Transwell assay showed that miR-27a inhibited FLS migration and invasion. However, miR-27a inhibition promoted the migration and invasion of FLS. In addition, the down-regulated expression of matrix metalloproteinases (MMP2, MMP9, and MMP13) and Rho family proteins (Rac1, Cdc42, and RhoA) was detected after treatment with miR-27a in RA-FLS by quantitative reverse transcription-PCR and western blot analysis. Then, a luciferase reporter assay validated that miR-27a targeted the 3-untranslated region (3'-UTR) of FSTL1. Moreover, miR-27a caused a significant decrease of FSTL1. In addition, the expression of TLR4 and $NF{\kappa}B$ was inhibited by miR-27a but increased by FSTL1 overexpression. In conclusion, we found that miR-27a inhibited cell migration and invasion of RA-FLS by targeting FSTL1 and restraining the $TLR4/NF{\kappa}B$ pathway.

• 동국한의학연구소논문집
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• 제7권2호
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• pp.107-120
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• 1999

본 연구는 관절염 유발시 일어나는 관절낭의 형태학적 변화를 조사하기위해 lipopolysaccharide(LPS) 주사로 인위적 관절낭 염증을 유발시킨 후 시간경과에 따른 윤활관절막과 섬유관절막의 향태 변화를 관찰하였다. BALB/C 암컷 생쥐 오른쪽 무릎관절낭에 LPS $300{\mu}g/kg$를 주사한 후 3, 7 그리고 14일에 무릎관절을 얻었다. 무릎관절은 4주동안 EDTA용액에 탈회한 후 통상적 방법으로 paraffin에 포매하였다. 또한 윤활관절막의 미세구조변화는 embed812로 포매한 후 관찰하였다. LPS 주사후 관절연골인접부위의 윤활관절막에서 시작된 세포과형성(hyperplasia)은 시간 경과후 전체 윤활관절막으로 확대되었다. 윤활관절막내의 미세구조의 변화로는 윤활포식세포(type I)가 관절강내로의 많은 돌기(filopodia)를 내었고, 잘 발달된 과립형질내세망을 가지는 type 2 윤활분비세포의 숫적 증가가 보였다. 한편 LPS 주사후 섬유관절막에서 나타나는 형태학적 변화는 collagen fiber 생성에 의한 섬유화가 증가되며, 이러한 섬유화를 주도하는 섬유모세포의 이주증가가 관찰되었다. 또한 혈관 주위에서는 백혈구의 이주 증가가 나타났으며, 탈과립형(degranulated type) 비만세포가 많이 관찰되었다. 이상의 결과로 LPS 주사로 관절낭에서 염증이 유발되어 윤활관절막과 섬유관절막에서 헝태학적 변화가 나타났다. 이러한 일련의 형태학적 변화는 발병초기 류마티스성 관절염에서 나타나는 병리학적 소견과 동일한 결과로서, 앞으로 진행될 치료제 개발과 유발기전에 관한 해석을 위한 in vivo 실험의 적절한 모델로 기여할 것으로 기대된다.

• IMMUNE NETWORK
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• 제7권1호
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• pp.39-47
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• 2007

Background: Stromal cell-derived factor (SDF)-1 is a potent chemoattractant for activated T cells into the inflamed Rheumatoid arthritis (RA) synovium. To determine the effect of macrophage migration inhibitory factor (MIF) on the production of SDF-1 in the inflamed RA synovium. Methods: The expression of SDF-1 and MIF in RA and Osteoarthritis (OA) synovium was examined by immunohistochemical staining. The SDF-1 was quantified by RT-PCR and ELISA after RA fibroblast like synoviocyte (FLS) were treated with MIF in the presence and absence of inhibitors of intracellular signal molecules. The synovial fluid (SF) and serum levels of MIF and SDF-1 in RA, OA and healthy control were measured by ELISA. Results: Expression of SDF-1 and MIF in synovium was higher in RA patients than in OA patients. The production of SDF-1 was enhanced in RA FLS by MIF stimulation. Such effect of MIF was blocked by the inhibitors of NF-${\kappa}B$. Concentrations of SDF-1 in the serum and SF were higher in RA patients than in OA patients and healthy control. SDF-1 and MIF was overexpressed in RA FLS, and MIF could up-regulate the production of SDF-1 in RA FLS via NF-${\kappa}B$-mediated pathways. Conclusion: These results suggest that an inhibition of interaction between MIF from T cells and SDF-1 of FLS may provide a new therapeutic approach in the treatment of RA.

• IMMUNE NETWORK
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• 제6권1호
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• pp.33-42
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• 2006

Background: Calcineurin plays a crucial role in T cell activation, cell growth, apoptosis, and angiogenesis, and its over-expression has been implicated in the pathogenesis of cardiomyopathy and stroke. However, the expression and function of calcineurin in the pathologic lesion of chronic inflammatory diseases, like rheumatoid synovium, remain to be defined. This study was aimed to determine the role of calcineurin in inflammatory arthritis and investigate the expression and function of calcineurin in the rheumatoid synovium and synoviocytes, the actual site of chronic inflammation. Methods: Immuno-histochemical staining using specific antibody to calcineurin was perfomed in the synovium of rheumatoid arthritis (RA). Fibroblast-like synoviocytes (FLS) from RA and osteoarthritis (OA) patients were isolated from RA and OA patients, and cultured with IL-1${\beta}$ and TNF-${\alpha}$ in the presence or absence of cyclosporin A, a calcineurin inhibitor. The calcineurin expression was assessed by phosphatase assay and Western blotting analysis. IL-6, -10, -17, matrix metalloproteinase (MMP)-1, -2, -3, and -9 released into the culture supernatants were measured by ELISA. After transfection with GFP-Cabin 1 gene into synoviocytes, the levels of IL-6 and MMPs were measured by ELISA. Results: Calcineurin was highly expressed in the lining layer of synovium and cultured synoviocytes of RA patients. The elevated calcineurin activity in the rheumatoid synoviocytes was triggered by proin flammatory cytokines such as IL-1${\beta}$ and TNF-${\alpha}$. In contrast, IL-10, an anti-inflammatory cytokine, failed to increase the calcineurin activity. The targeted inhibition of calcineurin by the over-expression of Cabin 1, a natural calcineurin antagonist, inhibited the production of IL-6 and MMP-2 by rheumatoid synoviocytes in a similar manner to the calcineurin inhibitor, cyclosporin A. Conclusion: These data suggest that abnormal activation of calcineurin in the synoviocytes may contribute to the pathogenesis of chronic arthritis, and thus provide a potential target for controlling inflammatory arthritis.

• IMMUNE NETWORK
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• 제8권1호
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• pp.29-37
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• 2008

Interleukin-23 (IL-23) is a novel pro-inflammatory cytokine which has been implicated to play a pathogenic role in rheumatoid arthritis (RA). This study was undertaken to investigate the IL-23 inductive activity of the proinflammatory cytokine IL-17, $IL-1{\beta}$ and tumor necrosis factor (TNF-${\alpha}$) in RA synovial fluid mononuclear cells (SFMC). Expression of IL-23p19, IL-17, $IL-1{\beta}$ and TNF-${\alpha}$ in joint was examined by immunohistochemistry (IHC) of patients with RA and osteoarthritis (OA). The effects of IL-17 and $IL-1{\beta}$ on expression of IL-23p19 in human SFMC from RA patients were determined by reverse transcriptase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). IL-23p19 was expressed in the RA fibroblast like synoviocyte (FLS), but not from OA FLS. Similar to the protein expression, IL-23p19 mRNA could be detected by RT-PCR in RA SFMC. IL-17 and $IL-1{\beta}$ could induce RA SFMC to produce the IL-23p19. The effects of IL-17 were much stronger than $IL-1{\beta}$ or TNF-${\alpha}$. These responses were observed in a doseresponsive manner. In addition, IL-17 or $IL-1{\beta}$ neutralizing antibody down-regulated the expression of IL-23p19 induced by LPS in RA-SFMC. Our results demonstrate that IL-23p19 is overexpressed in RA synovium and IL-17 and $IL-1{\beta}$ appears to upregulate the expression of IL-23p19 in RA-SFMC.