• IMMUNE NETWORK
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• 제2권1호
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• pp.25-34
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• 2002

Background: Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by synovial hyperplasia and joint destruction. The synovial fibroblasts express cell adhesion molecules and have a role in adhesive interation with inflammatory cells in synovial tissue. It has been suggested that hypoxic conditioins are thought to exist in arthritic joints, and several studies indicate that reactive oxygen species (ROS) produced in hypoxic condition can initiate events that lead to pro-adhesive changes via increased expression of adhesion molecules. So, this study wsa designed to examine whether antioxidant can inhibit hypoxia-induced expression of ICAM-1 in cultured human synovial fibroblasts. Methods: Synovial fibroblasts were isolated from synovial tissue in patients with RA and cultured at hypoxic condition. Antioxidant, PDTC (pyrrolidine dithiocarbamate) were pre-treated for an hour before the hypoxic culture and synovial fibroblasts were harvested at 0, 6, 12, 24, 48 hours time points. Cell surface ICAM-1 expression in synovial fibroblasts was examined by the flow cytometric analysis. To analyse the expression of ICAM-1 mRNA, reverse-transcriptase polymerase chain reaction (RT-PCR) was performed. The levels of cytokines in culture supernatants were measured by ELISA, and activation of NF-${\kappa}B$ was analysed by electrophoretic mobility shift assay. The adhesive reaction between synovial fibroblasts and lymphocytes was assayed by measurement of fluorescent intensity of BCECF-AM in lymphocytes. Results: Hypoxic stimuli up-regulated the ICAM-1 expression as well as the adhesive interaction of human synvial fibroblasts to lymphocytes in a time-dependent manner, and PDTC inhibited hpyoxia-induced ICAM-1 expression and cell-cell interaction. PDTC also inhibited the hypoxia-induced activation of intracellular transcription factor, NF-${\kappa}B$. PDTC decreased the amount of hypoxia-induced production of IL-$1{\beta}$ and TNF-${\alpha}$. Conclusion: These studies demonstrate that PDTC inhibit the hypoxia-induced expression of the adhesion molecule, ICAM-1 and activation of NF-${\kappa}B$ in cultured human synovial fibroblasts.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2001년도 추계학술대회 및 정기총회
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• pp.14-20
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• 2001

Rheumatoid arthritis (RA) is one of the most typical rheumatic diseases, and is characterized by chronic inflammation, cartilage destruction and joint deformity ［1,2］. During this process, profound hypertrophic changes of the synovium with infiltration of immune cells, increased vascularity, and hyperplasia result in the formation of a synovial pannus that invades cartilage and bone ［3］. In early stages of RA, the synovial membrane begins to invade the cartilage. In established RA, the synovial membrane becomes transformed into inflammatory tissue, the pannus (Fig. 1). The cell types that occupy cartilage-pannus junctions include synovial macrophages, fibroblasts, mast cells, polymorphonuclear lymphocytes (PMNs), and displaced, probably differentiated condrocytes ［4-6］. Recent studies of rheumatoid synovial tissue have demonstrated localized accumulations of mast cells and evidence of their activation/degranulation［7］.

• Journal of Microbiology and Biotechnology
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• 제18권4호
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• pp.686-694
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• 2008

The inhibitory effects of 5,6,3',5'-tetramethoxy 7,4'-hydroxyflavone (labeled as p7F) were elucidated on the productions of proinflammatory cytokines as well as inflammatory mediators in human synovial fibroblasts and macrophage cells. p7F inhibited IL-1${\beta}$ or TNF-${\alpha}$ induced expressions of inflammatory mediators (ICAM-1, COX-2, and iNOS). p7F also inhibited LPS-induced productions of nitric oxide and prostaglandin $E_2$ in RAW 264.7 cells. In order to investigate whether p7F would inhibit IL-1 signaling, p7F was added to the D10S Th2 cell line (which is responsive to only IL-1${\beta}$ and thus proliferates), revealing that p7F inhibited IL-1${\beta}$-induced proliferation of D10S Th2 cells in a dose-response manner. A flow cytometric analysis revealed that p7F reduced the intracellular level of free radical oxygen species in RAW 264.7 cells treated with hydrogen peroxide. p7F inhibited IkB degradation and NF-${\kappa}$B activation in macrophage cells treated with LPS, supporting that p7F could inhibit signaling mediated via toll-like receptor. Taken together, p7F has inhibitory effects on LPS-induced productions of inflammatory mediators on human synovial fibroblasts and macrophage cells and thus has the potential to be an anti-inflammatory agent for inhibiting inflammatory responses.

• Biomolecules & Therapeutics
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• 제28권2호
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• pp.172-183
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• 2020

Phosphoinositide 3-kinase (PI3K) is considered as a promising therapeutic target for rheumatoid arthritis (RA) because of its involvement in inflammatory processes. However, limited studies have reported the involvement of PI3KC2γ in RA, and the underlying mechanism remains largely unknown. Therefore, we investigated the role of PI3KC2γ as a novel therapeutic target for RA and the effect of its selective inhibitor, PBT-6. In this study, we observed that PI3KC2γ was markedly increased in the synovial fluid and tissue as well as the PBMCs of patients with RA. PBT-6, a novel PI3KC2γ inhibitor, decreased the cell growth of TNF-mediated synovial fibroblasts and LPS-mediated macrophages. Furthermore, PBT-6 inhibited the PI3KC2γ expression and PI3K/AKT signaling pathway in both synovial fibroblasts and macrophages. In addition, PBT-6 suppressed macrophage migration via CCL2 and osteoclastogenesis. In CIA mice, it significantly inhibited the progression and development of RA by decreasing arthritis scores and paw swelling. Three-dimensional micro-computed tomography confirmed that PBT-6 enhanced the joint structures in CIA mice. Taken together, our findings suggest that PI3KC2γ is a therapeutic target for RA, and PBT-6 could be developed as a novel PI3KC2γ inhibitor to target inflammatory diseases including RA.

• 한국동물학회:학술대회논문집
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• 한국동물학회 2001년도 한국생물과학협회 학술발표대회
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• pp.258.2-258
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• 2001

No Abstract, See Full Text

• IMMUNE NETWORK
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• 제7권1호
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• pp.10-17
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• 2007

Autoimmune arthritis, such as rheumatoid arthritis (RA), is a chronic inflammatory disorder that primarily affects the joints and then results in their progressive destruction. Effector Th cells have been classified as Th1 and Th2 subsets based on their cytokine expression profiles and immune regulatory function. Another subset of T cells termed Th17 was recendy discovered and known to selectively produce IL-17. Also, Th17 was shown to be generated by TGF${\beta}$ and IL-6 and maintained by IL-23. IL-17 is a proinflammatory cytokine that is considered to involve the development of various inflammatory autoimmune diseases such as RA, asthma, lupus, and allograft rejection. IL-17 is present in the sera, synovial fluids and synovial biopsies of most RA patient. IL-17 activates RA synovial fibroblasts to synthesize IL-6, IL-8 and VEGF via PI3K/Akt and NF-${\kappa}B$ dependent pathway. IL-17 increases IL-6 production, collagen destruction and collagen synthesis. In addition, it not only causes bone resorption but also increases osteoclastogenesis and fetal cartilage destruction. Inhibition of the IL-17 production may contribute a novel therapeutic approach along with potent anti-inflammatory effect and with less immunosuppressive effect on host defenses.

• Applied Microscopy
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• 제30권2호
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• pp.213-232
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• 2000

슬관절은 발생초 중간대에서 관절원판, 관절내인대 및 활액막세포들이 분화, 발육한다. 최근 관절경을 이용한 수술이 활발히 진행되고 있는 이때 관절내 구조들의 재생과정을 이해하기 위하여 이들의 발육과정을 전자현미경으로 관찰함은 매우 의의 있는 일이라 생각된다. 본 연구는 태생 초기부터 말기까지의 인태아를 대상으로 슬관절의 관절강 발생에서 중간대의 중간층세포의 운명과 활액막, 관절원판 및 관절내인대의 분화, 발육과정을 전자현미경으로 관찰하여 다음과 같은 결과를 얻었다. 좌고 30mm(태령 9주)때 십자인대와 관절원판의 원기들이 출현하였고 초기 관절강은 40mm(태령 10주)때 중간대의 중간층이 연골형성층에 결합하여 출현하였다. 반월연골은 60mm(태령 12주)때 간엽세포에서 섬유모세포로 분화하였고 100mm(태령 15주)때 섬유모세포는 내측에서는 난원형을, 중간과 외측에서는 장타원형을 각각 취하였다. 260mm(태령 30주)때 교원섬유의 주행은 내측에서 평행히 달리고 중간에서는 평행 또는 방사상 방향을, 외측에서는 상호교차 방향을 하고 있었다. 십자인대는 30mm때 섬유모세포로 분화하였고 세포질에 조면내형질망과 사립체를 함유하고 있었다. 교원원섬유는 섬유모세포의 돌기내에서 출현하였다. 100mm때는 교원 원세포는 세포의 간격을 가득 채우고 있었다. 260mm때 섬유모세포는 난원형 또는 방추상 핵을 가지고 있었고 교원섬유를 싸고 있지 않았으며 교원원섬유는 평행 또는 파상 방향으로 달리고 있었다. 활액막은 관절강의 초기 출현과 동시에 발생하였고 관절강이 확대되는 60mm때는 A와 B세포로 구분되었다. 100 mm때 활액막세포는 B세포가 대다수였고 B세포는 세포질에 다량의 조면내형질망과 잘 발달된 골지복합체를 함유하고 있었고 A세포는 사상위족, 음소포, 용해소체 및 공포를 가지고 있었다. 260mm에도 B세포가 대다수 관찰되었다. 같은 결과를 얻었다. 1. Estradiol benzoate를 4일간 매일 일정량을 투여한 흰쥐의 난관섬모세포내 $\alpha-tubulin$의 면역반응은 estradiol 투여후 1일, 3일 및 5일군에서 강한 반응을 나타내었다. 2. Estradiol beozoate를 4일간 투여한 후 cia-platin을 투여한 흰쥐 난관섬모세포내 $\alpha-tubulin$ 반응은 cis-platin 투여 1일군과 3일군에서 약한 반응을 나타내었으나 제 5일군에서는 강한 반응으로 회복되었다. 3. Cis-platin투여한 후 제 1일 및 3일군의 흰쥐 난관 섬모세포내 $\alpha-tubulin$반응은 첨부세포질에서는 감소되었고 기저체, 섬모등에서는 $\alpha-tubulin$반응이 대조군과 비교하면 변동이 없었다. 이상의 결과로 미루어 난관섬모세포내 $\alpha-tubulin$은 cis-platin투여에 의해 감소되는 것으로 결론 지을 수 있었다.調節) 작용(作用)에 중요한 역할을 하는 것이라 사료된다. 해수(海水)에 적응(適應)된 대부분의 틸라피아는 신장(腎臟)의 보우만 주머니를 가득 채운 신사구체(腎絲球體)를 가지고 있으며, 신사구체(腎絲球體)의 수축은 담수(淡水)에 적응된 개체보다 10%o, 20%o, 30%o에 적응된 개체에서 훨씬 더 많이 발생되었고, 울혈현상(鬱血現象)은 10%o보다 20%o, 30%o에 적응(適應)된 개체의 신장조직(腎臟組織)에서 더많이 발생되었다. 틸라피아의 신사구체(腎絲球體)는 담수(淡水)에서 10%o의 해수(海水)로 이주된지 14일(日) 이후에 신장(腎臟)에서 수축된 것으로 나타났다. 30%o의 해수(海水)에 적응(適應)된 틸라피아의 평균 신사구체(腎絲球體)의 면적은 담수(淡水)에 적응된 개체의 면적보다

• Korean Journal of Acupuncture
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• 제22권2호
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• pp.89-105
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• 2005

Objective & Methods : This study is performed to observe the effect of Herbal-acupuncture with Notopterygii Radix Herbal-Acupuncture Solution(NR-HAS) at Joksamni(ST36) on Collagen II-induced arthritis (CIA) in DBA/1J mice. Result : 1. The highest survival rate of mice lung fibroblasts were measured in the 1% NR-HAS, and the expression of $TNF-{\alpha}$ in synovial cells were significantly decreased in the 1% and 10% NR-HAS. 2. The incidence of arthritis and the spleen weight were significantly decreased by Notopterygii Radix Herbal-acupuncture(NR-HA) at ST36. 3. The levels of IL-6, $INF-{\gamma},\;TNF-{\alpha}$, IgG, IgM, anti-collagen II in serum of CIA mice were significantly decreased by NR-HA at ST36. 4. In histology, the cartilage destruction and synovial cell proliferation were decreased by NR-HA at ST36, and the collagen fiber expressions in the NR-HA I II groups were similar with that of the normal group. 5. In lymph node, the expression ratios of $CD3e^+\;to\;CD19^+$ cell and $CD4^+\;to\;CD8^+$ cell in the NR-HA I II groups were similarly maintained as those in the normal group. 6. In lymph node, $CD69^+/CD3e^+$ cells and $CD11a^+/CD19^+$ cells were decreased by NR-HA at ST36. 7. In the articular joint, $CD11b^+/Gr-1^+$ cells were decreased by NR-HA at ST36. 8. NR-HA at ST36 did not make a considerable difference in DBA/1J mice without CIA 9. Throughout the overall experimental result, NR-HA I group showed more predominant effect than the NR-HA II group. Conclusion : These results suggest that NR-HA at ST36 has an effect to control synovial cell proliferation and cartilage destruction in rheumatoid arthritis, as well as prophylaxis is important to treat rheumatoid arthritis in clinic.

• Journal of Acupuncture Research
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• 제23권3호
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• pp.129-142
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• 2006

Objectives : The purpose of this study is to observe the effects of Eucomiae Cortex herbal-acupuncture solution(EC-HAS) at Joksamni(ST36) on arthritis of mice induced by Collagen II. Methods : The author performed several experimental items. First, it is the cell survival rate of mice lung fibroblasts. Second, it is the incidence rate of arthritis and arthritis index of CIA. Third, it is the levels of IL-6, $TNF-{\alpha}$, $IFN-{\gamma}$, $IL-{\beta}$, IgG, IgM and anti-collagen II in serum and the level of IFN-y,$IFN-{\gamma}$/IL -4 ratio in CIA mouse spleen cell culture. Fourth, it is histological analysis of the mice joint. Fifth, it is expression ratio of $CD3e^+$ to $CD19^+$+ cell, $CD4^+$ to $CD8^+$ cell, $CD69^+/CD3e^+$/cells, $CD11a^+/CD19^+$/cells, $CD11b^+/Gr-1^+$ cells and $CD4^+/CD25^+$ cells. Results & Conclusion : 1. In the EC-HA, the incidence of arthritis and arthritis index were significantly decreased. 2. In EC-HA, the levels of IL-6, $IFN-{\gamma}$, $TNF-{\alpha}$, $IL-1{\beta}$, IgG, IgM and anti-collagen II in serum of CIA mice and the level of $IFN-{\gamma}$, IL-4, $IFN-{\gamma}$/lL-4 ratio in CIA mouse spleen cell culture were significantly decreased. 3. In the histological study, the cartilage destruction and synovial cell proliferation were decreased in the EC-HA, and the collagen fiber expressions in the EC-HA were similar with that of the Normal group. 4. In the EC-HA, the expression ratio of $CD3e^+$ to $CD19^+$ cell and $CD4^+$ to $CD8^+$ cell were similarly maintained as Normal group in lymph nodes, and $CD69^+/CD3e^+$ cells and $CD11a^+/CD19^+$ cells were decreased in lymph nodes, and $CD11b^+/Gr-1^+$ cells and $CD4^+/CD25^+$ cells were decreased in synovium. These results suggest that EC-HA at ST36 has an effect to control synovial cell proliferation and cartilage destruction in rheumatoid arthritis, and to be put to practical use in the future rheumatoid arthritis clinic.

• Journal of Acupuncture Research
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• 제23권6호
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• pp.29-44
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• 2006

Objectives : The purpose of this study is to observe the effects of Mori Ramulus herbal-acupuncture solution(MR-HAS) on arthritis of mice induced by Collagen II at Joksamni(ST36). Methods : The author performed several experimental items. First, it is the cell survival rate of mice lung fibroblasts. Second, it is the incidence rate of arthritis and arthritis index of CIA. Third, it is the levels of IL-6, $TNF-{\alpha}$, $IFN-{\gamma}$, $IL-1{\beta}$, IgG, IgM and anti-collagen II in serum and the level of $IFN-{\gamma}$, $IFN-{\gamma}/IL-4$ ratio in CIA mouse spleen cell culture. Fourth, it is histological analysis of the mice joint. Fifth, it is expression ratio of CD3e+ to CD19+ cell, CD4+ to CD8+ cell, CD69+/CD3e+ cells, CD11a+/CD19+ cells and CD11b+/Gr-l+ cells and CD4+/CD25+ cells. Results : 1. In the MR-HA, the incidence of arthritis and the arthritis index were significantly decreased. 2. In MR-HA, the levels of IL-6, $IFN-{\gamma}$, $TNF-{\alpha}$, $IL-1{\beta}$, IgG, IgM and anti-collagen II in serum of CIA mice and the level of $IFN-{\gamma}$, IL-4, $IFN-{\gamma}$, IL-4 ratio in CIA mouse spleen cell culture were significantly decreased. 3. In histology, the cartilage destruction and synovial cell proliferation were decreased in the MR-HA, and the collagen fiber expressions in the MR-HA were similar with that of the Normal group. 4. In the MR-HA, the expression ratio of CD3e+ to CD19+ cell and CD4+ to CD8+ cell were similarly maintained as Normal group in lymph nodes, and CD69+/CD3e+ cells and CD11a+/CD19+ cells were decreased in lymph nodes, and CD11b+/Gr-1+ cells and CD4+/CD25+ cells were decreased in synovium. Conclusion : These results suggest that MR-HA at ST36 has an effect to control synovial cell proliferation and cartilage destruction in rheumatoid arthritis, as well as prophylaxis is important to treat rheumatoid arthritis in clinic.