• Korean Journal of Pharmacognosy
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• v.36 no.4 s.143
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• pp.338-343
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• 2005

Inflammatory mediators such as interleukin-1 (IL-1), tumor necrosis $factor-{\alpha}\;(TNF-{\alpha}),\;interferon-{\gamma}\;(IFN-{\gamma})$ and lipopolysaccharide (LPS) are thought to play major roles in joint diseases such as a rheumatoid arthritis (RA), and there is considerable evidence playing a role for these cytokines in osteoarthritis (OA). Therefore, we have studied the effects on anti-inflammation and analgesic by ethyl acetate fraction from 70% ethanol extract of Artemisia capillaries (EAC). As a positive control, apigenin, which is known as an anti-inflammatory agent as an iNOS inhibitor, was used and showed the dose-dependent inhibitory effect. EAC showed strong inhibitory efficacy against cytokine-induced proteoglycan degradation, $PGE_2$ production, nitric oxide (NO) production, and matrix-matalloproteinases (MMPs) expression in rabbit articular chondrocyte. In the writhing test induced by acetic acid, EAC $(200{\sim}400\;mg/kg)$ exhibited a dose-dependent inhibition of writhing. The results indicate that EAC have anti-inflammatory and analgesic activities, and could be a good herbal medicine candidate for curing of RA and/or OA.

• Journal of Acupuncture Research
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• v.25 no.1
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• pp.25-55
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• 2008

Objectives : The aim of the present study was to observe the effect of Astragali Radix Herbal-Acupuncture Solution(AR-HAS) at $ST_{36}$(jok-samni, $Z\acute{u}s\bar{a}n$ Li) on collagen- II -induced arthritis(CIA) in mice. Methods : DBA/1J mice were immunized with bovine type II collagen(CII) on days 0 and 21 to induce arthritis. The mice were divided into 5 groups : normal group(no CIA), control group(CIA+no treatment), needle prick group(CIA+single prick with an injection needle), saline group(CIA+saline injection) and ARHA group(CIA+ R-HA treatment). The needle prick, saline injection, and AR-HA groups were injected on the right $ST_{36}$(jok-samni) of mice for 9 weeks, 3 times a week, beginning 4 weeks after the booster immunization. Results : 1. The incidence of arthritis, AI(arthritis index), and joint edema decreased in the AR-HA group. 2. Weight gain, hypertrophy of the spleen, adhesion of the tissues, and transformation of the joint were restrained in the AR-HA group. 3. The concentrations of $IL-1{\beta}$, IL-6, $TNF-{\alpha}$, $IFN-{\gamma}$ in CIA mouse serum and $IFN-{\gamma}$, IL-10 in the CIA mouse spleen cell culture decreased significantly at $ST_{36}$ in the AR-HA group. 4. Total cell counts increased significantly, and the ratio of $CD3e^+$ to $CD45R^+$, $CD4^+$ to $CD8^+$, and $CD4^+$ to $CD25^+$ cells decreased significantly at $ST_{36}$ in the CIA mouse spleen cell culture of the AR-HA group. 5. Total cell counts decreased significantly, and $CD4^+/CD25^+$ and $CD45R^+/CD69^+$ decreased significantly at $ST_{36}$ in the CIA mouse lymph nodes of the AR-HA group. 6. $CD3e^+/CD69^+$ and $CD11b^+/Gr-1^+$ cells decreased significantly at $ST_{36}$ in the CIA mouse joints of the AR-HA group. 7. The histological examination showed that cartilage destruction and synoviocyte proliferation decreased in the CIA mouse joints of the AR-HA group, and collagen fiber was expressed similar to that seen in the normal group. Conclusions : Our experiments show that at $ST_{36}$, an anti-inflammatory mechanism of AR-HA controls synovial cell proliferation and protects against cartilage destruction in rheumatoid arthritis.

• The Korean Journal of Physiology and Pharmacology
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• v.8 no.5
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• pp.273-280
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• 2004

Nitric oxide (NO) has been suggested to act as a mediator of cytokine-induced effects of turn over of bone. Activation of the inducible nitric oxide synthase (iNOS) by inflammation has been related with apoptotic cell death in osteoblast. YS 49, a synthetic isoquinoline alkaloid, inhibits NO production in macrophages activated with cytokines. In the present study, we investigated the molecular mechanism of YS 49 to inhibit iNOS expression in ROS 17/2.8 cells, which were activated with combined treatment of inflammatory cytokines $(TNF-{\alpha},\;IFN-{\gamma})$ and lipopolysaccharide (LPS). Results indicated that YS 49 concentration-dependently reduced iNOS mRNA and protein expression, as evidenced by Northern and Western blot analysis, respectively. The underlying mechanism by which YS 49 suppressed iNOS expression was not to affect iNOS mRNA stability but to inhibit activation and translocation of $NF-_kB$ by preventing the degradation of its inhibitory protein $I_kB_{\alpha}$. As expected, YS 49 prevented NO-induced apoptotic cell death by sodium nitroprusside. Taken together, it is concluded that YS 49 inhibits iNOS expression by interfering with degradation of phosphorylated inhibitory $_kB_{\alpha}\;(p-I_kB_{\alpha})$. These actions may be beneficial for the treatment of inflammation of the joint, such as rheumatoid arthritis.

• Journal of Acupuncture Research
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• v.17 no.1
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• pp.221-250
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• 2000

In order to investigate experimentally that the effect of aqua-acupuncture solution of Paeonia lactiflora on arthritis of mice induced by collagenII, the author performed several experimental items : that is increase, paw thickness, DTH, weight of spleen, hematological change, expression of $CD4^+$, $CD8^+$, $CD19^+$, gene expression of IL-1, IL-6, IL-12, IFN-${\gamma}$, TNF, proliferation of synovial cells and cytotoxicity. The results were obtained as follows; 1. Inhibitory effects of aqua-acupuncture solution of Paeonia lactiflora on arthritis induced by collgenII. 1) In incidence, paw edema, AI and DTH were inhibited as compared with control group. 2) The splenic weight was increased and the number of leukocytes was decreased as compared with control group. 3) The number of $CD4^+$, $CD8^+$ activated cells and surface-receptor expression were increased as compared with control group. 4) In hematological change, total protein, creatinine and LDH were decreased significantiy as compared with control group. 2. FACS analysis on normal BABL/c of spenic cells treated with aqua-acupuncture solution of Paeonia lactiflora. 1) Aqua-acupuncture solution of Paeonia lactiflora activated adhesive splenic cells of mice morphologically in vitro. 2) Aqua-acupuncture solution of Paeonia dactiflora enhanced the gene expression of IL-12 and also enhanced that of interferon-${\gamma}$ remarkably. 3) Aqua-acupuncture solution of Paeonia lactiflora reduced the number of $CD4^+$, $CD8^+$, $CD19^+$ activated cells and their surface-receptor expression as compared with control group. 3. Effects of aqua-acupuncture solution of Paeonia lactiflora on human synovial cells. 1) In cytotoxicity against synovial cells, aqua-acupuncture solution of Paeonia lactiflora didn't show cytotoxicity at concentration of $10-100{\mu}g/m{\ell}$ but showed significantly at concentration of $200-400{\mu}g/m{\ell}$ as compared with control group. 2) Aqua-acupuncture solution of Paeonia lactiflora reduced the gene expression of IL-6, IL-$1{\beta}$ and TNF-${\alpha}$. 3) Aqua-acupuncture solution of Paeonia lactiflora inhibited proliferation of synovial cells at concentration of 100 and $200{\mu}g/m{\ell}$.

• Journal of Acupuncture Research
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• v.17 no.1
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• pp.287-318
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• 2000

The purpose of this study is to observe the effect of Aqua-acupuncture utilizing Lycopi Herba Solution(LH-AS) on arthritis. For that purpose, we formed three experimental group, synovial cells of human body, normal BALB/C mice and DBA/1J mice with collagen II-induced arthritis, and measured the treatment effect of LH-AS on each group. The conclusions are as follows. 1. After the LH-AS treatment on synovial cells, there were no significant change in 1, 10, 50 and $100{\mu}g/m{\ell}$ whereas there was significant change in 200 and $400{\mu}g/m{\ell}$ in cytotoxicity. 2. IL-6, IL-$1{\beta}$, TNF-${\alpha}$ gene expression of synovial cells and the secretion amount of IL-6 and IL-$1{\beta}$ are significantly inhibited in treatment group with LH-AS. 3. The proliferation of synovial cells was significantly inhibited in treatment group with rIL-6, LH-AS 200 and rIL-6, $100{\mu}g/m{\ell}$. 4. After the DBA/1J mice with collagen II-induced arthritis were treated by LH-AS, the incidence of arthritis, hind paw edema, the index of arthritis and DTH were significantly inhibited. 5. After the DBA/1J mice with collagen II-induced arthritis were treated by LH-AS, splenetic weight was significantly increased and the number of leukocyte was significantly decreased. But there was no significant change in the number of platelet. 6. After the DBA/1J mice with collagen II-induced arthritis were treated by LH-AS, the number of $CD4^+$, $CD8^+$ activated cells and the surface-receptor expression were significantly increased whereas the number of $CD19^+$ activated cells and the surface-receptor expression were decreased. 7. After the DBA/1J mice with collagen II-induced arthritis were treated by LH-AS, total protein, LDH were significantly decreased, but there was no significant change in creatinine. 8. After the normal splenetic cells of BALB/C mice were treated by LH-AS and cultured, it was observed that the adherent cells were morphologically activated and IL-12 and IFN-${\gamma}$ gene expression were increased. 9. After the normal splenetic cells of BALB/C mice were treated by LH-AS and cultured, the number of $CD4^+$, $CD8^+$, $CD19^+$ activated cells and surface-receptor expression were inhibited when being compared with the control group. Taking all these observations into account, LH-AS injection is considered to be effective in treating arthritis and put to practical use in future arthritis clinic.

• IMMUNE NETWORK
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• v.3 no.1
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• pp.1-7
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• 2003

Type II collagen (CII), major component of hyaline cartilage, has been considered as an auto-antigen in rheumatoid arthritis (RA). However, the clinical and biological significances with regard to the CII autoimmunity need to be clarified in human RA. The presence of antibodies to CII has been identified in sera, synovial fluid, and cartilage of patients with RA. In our study, the increased titer of IgG anti-CII in sera was well correlated with C-reactive protein, suggesting that this antibody may reflect the inflammatory status of RA. The titer of anti-CII antibodies (anti-CII Abs) tended to be higher in early stages of diseases. In our extending study, among 997 patients with RA, 269 (27.0%) were positive for circulatory IgG antibody to CII, those levels were fluctuated over time. It is hard to assess the significant amount of T cell responses to CII and CII (255~274) in RA. By using a sensitive method of antigen specific mixed lymphocyte culture, we can detect the presence of CII-reactive T cells in peripheral blood mononuclear cells of RA patients. Sixty seven (46.9%) of 143 patients showed positive CII reactive T cell responses to CII or CII (255~274). The frequencies of CII reactive T cells were more prominent in inflamed synovial fluid (SF) than in peripheral blood. These T cells could be clonally expanded after consecutive stimulation of CII with feeding of autologous irradiated antigen presenting cells (APC). Moreover, the production of Th1-related cytokine, such as IFN-${\gamma}$, was strongly up-regulated by CII reactive T cells. These data suggest that T cells responding to CII, which are probably presenting the IFN-${\gamma}$ producing cells, may play an important role in the perpetuation of inflammatory process in RA. To evaluate the effector function of CII reactive T cells, we investigated the effect of CII reactive T cells and fibroblasts-like synoviocytes (FLS) interaction on the production of pro-inflammatory cytokines. When the CII reactive T cells were co-cultured with FLS, the production of IL-15 and TNF-${\alpha}$ from FLS were significantly increased (2 to 3 fold increase) and this increase was clearly presented in accord to the expansion of CII reactive T cells. In addition, the production of IFN-${\gamma}$ and IL-17, T cell derived cytokines, were also increased by the co-incubation of CII reactive T cells with FLS. We also examined the impact of CII reactive T cells on chemokines production. When FLS were co-cultured with CII stimulated T cells, the production of IL-8, MCP-1, and MIP-1${\alpha}$ were significantly enhanced. The increased production of these chemokines was strongly correlated with increase the frequency of CII reactive T cells. Conclusively, immune response to CII was frequently found in RA. Activated T cells in response to CII contributed to increase the production of proinflammatory cytokines and chemokines, which were critical for inflammatory responses in RA. The interaction of CII-reactive T cells with FLS further augmented this phenomenon. Taken together, our recent studies have suggested that autoimmunity to CII could play a crucial role not only in the initiation but amplification/perpetuation of inflammatory process in human RA.

• IMMUNE NETWORK
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• v.5 no.2
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• pp.78-88
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• 2005

Background: Collagen-induced arthritis (CIA) in mice is animal model of autoimmune disease known as rheumatic arthritis in human. We investigated CII-specific CD4+ T cell receptor usage in CIA mice. Methods: In CIA model, draining lymph node (dLN) CD4+ T cells and splenocytes at $3^{rd},\;5^{th},\;8^{th}$ week, we investigated CII-specific T cell proliferation, production of IL-17, IFN-${\gamma}$, TNF-${\alpha}$, IL-4 and IL-10. And we also performed anti-CII IgG Ab measurements in serum level, TCRV ${\beta}$ usage and T cell clonality with RT-PCR-SSCP analysis. Also, we performed proliferative response against CII when CII-specific T cell subset is deleted. Results: CIA mice showed more increase in the serum level of anti-CII IgG than normal mice after induction of arthritis. And the level of anti-CII IgG2a in CIA mice was increased after $3^{rd}$ week after primary immunization, while anti-CII IgG1 was decreased. Draining LN CD4+ T cells have proliferated against CII stimulation at $3^{rd}$ week after $1^{st}$immunization. CD4+T cells derived from dLN of CIA mice produced proinflammatory cytokine IFN-${\gamma}$, IL-17 etc. Draining LN CD4 T cells of CIA presented higher proportion of CD4+V ${\beta}3$+subset compared to those of normal mice at $3^{rd}$ week after $1^{st}$ immunization, and they were increased in proportion by CII stimulation. Draining LN CD4+ T cells without TCRV ${\beta}3+/V{\beta}8.1/8.2+/V{\beta}$10b+cells were not responsive against CII stimulation. But, CII-reactive response of TCRV ${\beta}3-/V{\beta}8.1/8.2-/V{\beta}$10b- T cells was recovered when $V{\beta}3+$ T cells were added in culture. Conclusion: Our results indicate that CD4+$V{\beta}3+$ T cells are selectively expanded in dLN of CIA mice, and their recovery upon CII re-stimulation in vitro, as well as the production Th1-type cytokines, may play pivotal role in CIA pathogenesis.