• Journal of Korean Medicine Rehabilitation
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• v.25 no.1
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• pp.27-44
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• 2015

Objectives This study was carried out to find the effects of Gami-cheongyulsaseub-tang (hereinafter referred to GCST) on the inhibition of zymosan-induced pain in rats and collagen II-induced arthritis (CIA) in DBA/1J mouse. Methods As an acute inflammatory pain model, peripheral inflammation was induced by intraplantar injection of zymosan into the right hind paw in rats and then the hyperalgesia and pain regulating factors in spinal cord were analyzed. As a chronic inflammation model, the mixture of collagen II and complete Freund's adjuvant was treated into mice to establish rheumatoid arthritis and then body weight, thickness of hind paw, pathological change of spleen, immunological rheumatoid factor (IgG1, IgG2a, IgG2b, IgM and anti-collagen II), pro-inflammatory cytokines, and bone injury were analyzed. Results In the acute inflammatory pain model, GCST significantly inhibited the thermal and mechanical hyperalgesia and the pain regulating factors, including Fos, CD11b, PKA and PKC, in the spinal cord with a dose-dependent manner. In the chronic rheumatoid arthritis model, GCST administration decreased arthritic index and paw edema as compared with CIA control group. In particular, GCST reduced significantly the serum levels of total IgG2a, IgG2b, IgM, and specific anti-collagen II, but not total IgG1. GCST also resulted in the attenuation of bone injury and spleen enlargement/adhesion in CIA mice. Moreover, the secretion of pro-inflammatory cytokines TNF-${\alpha}$ and IL-$1{\beta}$ in CIA mice was significantly reduced by GCST in a dose-dependent manner. Conclusions Comparison of the results in this study showed that GCST had anti-nociceptive and immunomodulatory effects. These data imply that GCST can be used as an effective drug for not only rheumatoid arthritic pain but also other auto-immune diseases.

• The Korean Journal of Physiology and Pharmacology
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• v.14 no.4
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• pp.199-204
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• 2010

The potential therapeutic action of shikonin in an experimental model of rheumatoid arthritis (RA) was investigated. As a RA animal model, DBA/1J mice were immunized two times with type II collagen. After the second collagen immunization, mice were orally administered shikonin (2 mg/kg) once a day for 35 days, and the incidence, clinical score, bone mineral density (BMD), bone mineral content (BMC) and joint histopathology were evaluated. BMD in the proximal regions of the tibia largely increased in the shikonin treatment group compared with the control group. We also examined the effect of shikonin on inflammatory cytokines and cartilage protection. Shikonin treatment significantly reduced the incidence and severity of collagen-induced arthritis (CIA), markedly abrogating joint swelling and cartilage destruction. Shikonin also significantly inhibited the production of matrix metalloproteinase (MMP)-1 and up-regulated tissue inhibitors of metalloproteinase (TIMP)-1 in mice with CIA. In conclusion, shikonin exerted therapeutic effects through regulation of MMP/TIMP; these results suggest that shikonin is an outstanding candidate as a cartilage protective medicine for RA.

• The Korean Journal of Physiology and Pharmacology
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• v.19 no.2
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• pp.83-88
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• 2015

CD4+CD25+ regulatory T cells (CD4+CD25+ Tregs) have been shown to play a regulatory or suppressive role in the immune response and are possibly relevant to the pathogenesis of autoimmune diseases. In the present study, we attempted to investigate the frequency of CD4+CD25+ Tregs in peripheral blood (PB) of collagen-induced arthritis (CIA) rats during the development of arthritis, to determine whether their frequency is involved in the immunoregulation of this disease. The results showed that normal rats had similar frequencies of CD4+CD25+ Tregs in PB during the experiment time, expressed as a percentage of CD4+CD25+Foxp3+ T cells among the CD4+ T lymphocyte population. In contrast, the frequency of CD4+CD25+Foxp3+ T cells in CIA rats was found to change during the development of arthritis. In CIA rats, there is a significant negative correlation between the frequency of CD4+CD25+Foxp3+ T cells and paw swelling (r=-0.786, p< 0.01). The relationship between the frequency of CD4+CD25+Foxp3+ T and immune activation was not found in normal rats. During the time course, the frequency of CD4+CD25+Foxp3+ T was lower in CIA rats than in normal ones. The data suggest that the frequency of PB CD4+CD25+ Tregs may be a promising marker for arthritis activity.

• Journal of Acupuncture Research
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• v.22 no.1
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• pp.99-108
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• 2005

Objective : The aim of this study is to investigate the analgesic effect and its mechanism of bee venom acupuncture on collagen-induced arthritis(CIA) rats. Methods : Bee venom (1 mg/kg) was subcutaneously punctured into Choksamni (ST36) of CIA Analgesic effect was evaluated by using the tail flick latency (TFL). Opioid and ${\alpha}2$-adrenergic neurotransmitter system were examined by naloxone as an opioid receptor antagonist and yohimbine as ${\alpha}2$-adrenoceptor antagonist prior to bee venom cupuncture. Results : The results were as follows; 1. The TFL for the CIA rat was decreased as time went by. 2. The TFL in CIA rat was increased in bee venom acupuncture group compared with control group (no treatment). 3. Analgesic effect of bee venom acupuncture was not abolished by naloxone pre-treatment in the CIA rat. 4. Analgesic effect of bee venom aqua-acupuncture was abolished by yohimbine pre-treatment in the CIA rat. 5. Two weeks bee venom acupuncture had the continous analgesic effect for 4 weeks. Conclusions : Bee venom acupuncture has an analgesic effect on the CIA rat and has an antinociception mediated by ${\alpha}2$-adrenergic system.

• Journal of Acupuncture Research
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• v.24 no.6
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• pp.15-27
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• 2007

Objectives : The purpose of this study is to investigate the effectiveness of Ulmus davidiana Planch herbal acupuncture(UA) to inhibit nuclear $factor(NF)-{\kappa}B$ activation on type II collagen-induced arthritis (CIA) in mice. Methods : Using an in vitro test, the synoviocytes picked out from the experimental CIA mice were subcultured. The synoviocyte cells were treated with phorbol-12-myristate-13-acetate(PMA) for 1 hour prior to the addition of indicated concentrations($0.4\;-\;1.0mg/m{\ell}$) of UA, and the cells were further incubated for 24 hours. The in vivo test, $NF-{\kappa}B$ p65, inducible nitric oxide synthase(iNOS), cyclooxygenase-2(COX-2), vascular cell adhesion molecule(VCAM)-1 production and apoptosis was observed by immunohistochemical staining. Results : The PMA-induced $I{\kappa}B$ kinase(IKK), iNOS and COX-2 mRNA expression were dose-dependently decreased in UA treated synoviocytes. Using the in vivo test, the number of eosinophils in mice treated with UA noticeably decreased in the the CIA group(P<0.05 using student t test). In mice treated with UA, there was less cartilage erosion. less bone destruction, mild synovial hyperplasia, mild fibrosis, and mild angiogenesis with less MIP-2 production. By immunohistochemical staining, suppression of $NF-{\kappa}B$ p65, iNOS production, inhibition of COX-2 production, inhibition of VCAM-1 production and inducing apoptosis were observed. Conclusions : These results suggest that UA might be applicable to the therapy of RA to suppress $NF-{\kappa}B$ activation.

• Journal of Acupuncture Research
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• v.19 no.5
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• pp.10-27
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• 2002

연구목적 : 면역억제와 활성 작용을 지닌 것으로 알려진 녹용약침(CPH)을 type II collagen 유발 관절염 (CIA) 백서와 인산이온 유발 연골세포의 세포사에 있어 보호활성 효과를 연구하였다. 연구방법 : 7주된 암컷 Sprague-Dawley 쥐를 collagen으로 관절염을 유발시킨 후 CPH의 효과를 관절염 점수, 체중감소 등의 평가기준으로 검정하였다. CPH는 일주일에 5번씩 각각 10, 20, 30 및 $100{\mu}g/kg/day$의 용량으로 양측 신수혈에 주입하였다. 연구결과 : 1. 300 mg/kg/day CPH처리로 관절염점수의 감소를 기초로 한 collagen 유발 관절염의 발생을 완전히 억제하였으며 관절염 점수상에서 CPH의 효과작인 용량은 64 mg/kg이었다. 2. CHP는 쥐의 간에 있는 DHO-DHase 활성을 $Ki=843{\pm}43{\mu}g/ml$의 비교적 높은 비활성으로 억제하였다. 3. 관절염관련 세포의 증식억제활성을 검정한 결과 CPH의 항 증식효과는 세포주기 S기에서 정지시키는 활성을 나타내었다. 4. 쥐의 늑연골로부터 완전히 최종 분화된 비대연골세포를 분리 배양하여 3~5mM/L Pi를 첨가함으로서 세포사멸을 측정하였다. $10{\mu}g/ml$ CPH 처리에 의한 보호(억제) 효과가 Pi-유발 연골세포의 세포사에 대한 Na-Pi cotransport의 경쟁적 저해제로 알려진 phosphonoformic acid(PFA)의 억제활성과 상응하는 수준으로 CPH의 활성을 확인하게 되었다.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.5
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• pp.1278-1284
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• 2007

Solanum Iyratum Thunb (Solanaceae) has multiple applications in korean traditional medicine because of its cytotoxic, immunological and anti-inflammatory activities. However, no study on the anti-arthritic activity of Solanum Iyratum Thunb has been reported in vivo. Rheumatoid arthritis (RA) is a systemic autoimmune disease with chronic inflammation characterized by hyperplasia of synovial cells in affected joints, which ultimately leads to the destruction of cartilage and bone. Cytokine production were assessed during CIA(collagen-induced arthritis) model mice in lymph node (LN), in knee joint and spleen, using ELISA. DBA/1j mice were immunized with bovine type II collagen. After a second collagen immunization, mice were treated with Solanum Iyratum Thunb (SLT) orally at 400, 200 mg/kg once a day for 4 weeks. The severity of arthritis within the knee joints was evaluated by histological assessment of cartilage destruction and pannus formation. SLT significantly suppressed the progression of CIA and inhibited the production of TNF-alpha and IFN-gamma in serum and spleen cell culture supernatant. The erosion of cartilage was dramatically reduced in mouse knees after treatment with SLT. In conclusion, our results demonstrates that SLT significantly suppressed the progression of CIA. This action was characterized by suppression of arthritis index, cartilage erosion and synovial cell infiltration and the decreased production of $TNF-{\alpha}$, $IFN-{\gamma}$, CD4+, CD19+, CD3+/CD69+ cells (in lymph node), CD11b+/Gr-1 + (in knee joint).

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.6
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• pp.1416-1422
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• 2008

In order to examine anti-inflammatory effect of Anemarrhenae Rhizoma (AR) alcohol extract on rheumatoid arthritis, the present study investigated the viability and TNF-${\alpha}$ production in Raw264.7 cells treated with AR and collagen induced arthritis in DBA/1J mice which were orally administered with AR prior to immunization. The results are as follows: AR extract at 20 and 50${\mu}g$/ml inhibited the viability of Raw264.7 by 35% and 79%, respectively. AR showed a significant decrease in TNF-${\alpha}$ levels from Raw264.7 cells treated with LPS. AR administration significantly decreased arthritic index in DBA/1J mice immunized with bovine collagen type II. AR administration significantly decreased spleen weights obtained from mice in 6 weeks after immunization. AR administration significantly decreased serum anti-type II collagen antibody levels compared with control group. AR administration decreased serum IL-6 levels compared with control group but it did not reach statistical significance.

• Journal of Acupuncture Research
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• v.18 no.1
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• pp.113-128
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• 2001

Objective : To investigate the effect of BUM aqua-acupuncture in treating the RA, the immunosis to logical analysis of LPS induced arthritis in mice to study this. For 14th day after the injection of LPS & BUM injection, the distribution of fibroblast, collagen, CD54(ICAM-1), CD106(VCAM-1), IL-$1{\beta}$, IL-2 receptor, CDl lb(macrophage) were examined on synovial capsule of mice knee joint. For 14th day after the injection of LPS & BUM injection, the distribucion of CD4(TH cell), CD8(TC cell), CD40(B cell) were examined on common iliac lymph node in mice. Methods : The experimental model of arthritis was induced by injection of 300${\mu}g$/kg LPS in BALB/c mice weighing 30g. The 100${\mu}l$ BUM aqua-acupuncture which compounded calculus bovis, fel ursi and moschus was injected into GB34 of mice every other day for 12 days. For 3rd, 7th, 14th day after the injection of LPS, the neutrophil, lymphocyte and monocytc counts in WBC were measured using hemacytometer. Results : The obstain results are summarized as follows ; 1. In sample group, the neutrophils counts were increased and the lympnocytes counts were decreased compared with control group. 2. The distribution of fibrosis & fibroblast on synovial membrane were decreased compared with control group. 3. The distribution of collagen fiber on synovial membrane were decreased compared' with control group. 4. The distribution of CD54(ICAM-1) & CD106(VCAM-1) on synovial membrane were decreased compared with control group. 5. The distribution of IL-$1{\beta}$ & IL-2 receptor on synovial membrane were decreased compared with control group. 6. The distribution of CDb(macrophage) on synovial membrane were decreased compared with control group. 7. The distribution of CD4(TH cell), CD8(TC cell) and CD40(B cell) in common iliac lymph nodes were decreased compared with control group. Conclusions : BUM aqua-acupuncture stimulation decreased inflammatory responses LPS induced arthritis in mice.

• Journal of Microbiology and Biotechnology
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• v.17 no.2
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• pp.348-358
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• 2007

Metallothionein, a cysteine-rich stress response protein that is naturally induced by a variety of immunologic stressors, has been shown to suppress autoimmune disorders through mechanisms not yet fully defined. In the present study, we examined the underlying mechanisms by which metallothionein might mediate such regulation of autoimmunity. $Na\ddot{i}ve\;CD4^+$ T cells from metallothionein-deficient mice differentiated to produce significantly less IL-10, $TGF-{\gamma}$, and repressor of GATA, but more $IFN-{\gamma}$ and T-bet, when compared with those from wild-type mice. The levels of IL-4 and GATA-3 production were not different between the two groups of mice. Conversely, treatment with exogenous metallothionein during the priming phase drove $na\ddot{i}ve$ wild-type $CD4^+\;T$ cells to differentiate into cells producing more IL-10 and $TGF-{\beta}$, but less $IFN-{\gamma}$ than untreated cells. Metallothionein-primed cells were hyporesponsive to restimulation, and suppressive to T cell proliferation in an IL-10-dependent manner. Lymphocytes from metallothionein-deficient mice displayed significantly elevated levels of AP-1 and JNK activities in response to stimulation compared with those from wild-type controls. Importantly, transgenic mice overexpressing metallothionein exhibited significantly reduced susceptibility to collagen-induced arthritis and enhanced IL-10 level in the serum, relative to their nontransgenic littermates. Taken together, these data suggest that metallothionein is able to promote the generation of IL-10-and $TGF-{\beta}$-producing type 1 regulatory T-like cells by downregulating JNK-dependent AP-1 activity. Thus, metallothionein may play an important role in the regulation of Th1-dependent autoimmune arthritis, and may represent both a potential target for therapeutic manipulation and a critical element in the diagnostic assessment of disease potential.