Journal of Physiology & Pathology in Korean Medicine
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v.22
no.4
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pp.871-877
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2008
Heme oxygenase-1 (HO-1), an inducible heme-degrading enzyme, is expressed by macrophages and endothelial cells in response to various stresses and mediators of inflammation. HO-1 has been recently implicated in regulation of angiogenesis via expression of VEGF. The purpose of this study was to determine the effects of HO-1 modulation on the collagen-induced arthritis (CIA) model and on angiogenesis via up- regulation of VEGF expression in human synovial fibroblast. DBA/1J mice were treated with an inhibitor of HO-1, tin protoporphyrin IX (SnPP), or with an inducer of HO-1, cobalt protoporphyrin IX (CoPP), from day 1 to day 35 after CIA induction. The clinical evolution of disease was monitored visually. At the end of the experiment, histopathologic changes were examined on the joints. VEGF expression in paws were measured by immunohistochemical stain. mRNA expression of HO-1 and VEGF stimulated with various concentration of $TNF-{\alpha}$, CoPP accessed on human synovial fibroblast by RT-PCR. Effects of pretreatment with SnPP on mRNA expression of HO-1 and VEGF in the presence of CoPP and $TNF-{\alpha}$ in synovial fibroblast was accessed by Real-time RT-PCR. Administration of cobalt protoporphyrin IX significantly induced the inflammatory response, with increased arthritis index and expression of VEGF in the paws of the arthritis models. Treatment with SnPP significantly reduced the severity of CIA through inhibition of joint inflammation and cartilage destruction. The expression of VEGF were also significantly reduced by SnPP treatment in the paw. CoPPIX as inducer of HO-1, increased HO-1 and VEGF expression dose dependently in synovial fibroblast. In contrast, inhibition of HO-1 activity by SnPPIX abrogated CoPPIX-induced HO-1 and VEGF production in synovial fibroblast. Stimulation with $TNF-{\alpha}$ increased HO-1 and VEGF expression itself and showed additive effect on HO-1 and VEGF expression when it treated with CoPP. When SnPP was treated with CoPP and $TNF-{\alpha}$, it abrogated the CoPP induced HO-1 and VEGF expression and also abrogated $TNF-{\alpha}$ induced HO-1 and VEGF expression in synovial fibroblast. The effects of HO-1 induction in rheumatoid arthritis results in aggravation of arthritis via up-regulation of VEGF. I concluded that inhibition of the expression or activity of HO-1 could be a therapeutic target of rheumatoid arthritis.
Objectives : The aim of the present study is to examine the effect and mechanism of Erycibae Caulis and Corydalis Tuber Pharmacopuncture (ECP) on a mouse model with collagen induced rheumatoid arthritis (CIA). Methods : We evaluated the Aspartate aminotransferase (AST), Alanine aminotransferase (ALT), Creatinine, and the Blood urea nitrogen (BUN) of serum to examine the safety of this study. In vivo, we compared the results of the non-treated group, the normal saline pharmacopuncture treated control group, the indomethacin treated group and the ECP group. We evaluated rheumatoid arthritis manifestation and the Rheumatoid Arthritis Index (AI). Also, immune cells in blood affected by ECP were evaluated by calculating the level of white blood cells (WBC), neutrophil, lympocytes and monocytes. Next, the level of Immunoglobulin M (IgM), Immunoglobulin G (IgG), Interleukin (IL)-$1{\beta}$, IL-6, IL-17, Tumor Necrosis Factor (TNF)-${\alpha}$ and Granulocyte-macrophage Stimulating Factor (GM-CSF)in serum were measured. We examined the imaging of cartilage degeneration using micro CT-arthrography of the hind paw. Additionally, we examined the effects of reducing bone volume (BV) ratio and bone surface/bone volume (BS/BV) ratio with 3D Micro-CT. Finally, we did a histopathologic examination analysis. Results : The absence of liver and kidney toxicity was evident. In vivo, edema of the joints of the ECP group decreased greatly in macroscopic observation. AI measurement of the ECP group also decreased significantly compared to the control group. The level of WBC, neutrophil, lympocytes, and monocytes in the blood decreased but there was no statistical significance of this data. IgM of the ECP group decreased significantly compared to the control group. IL-$1{\beta}$, IL-6, TNF-${\alpha}$, and GM-CSF production of the ECP group decreased significantly compared to the control group. As a result of examining joint condition with 3D micro CT, deformation and destruction of the joint was shown to have decreased. Bone density of ECP group increased at a statistically significant level compared to the control group. Degree of joint inflammation of ECP group decreased significantly compared to the control group. After H&E and M-T staining, infiltration of immune cells, subsidence of the cartilage, damage to the synovial cells and joint erosion decreased. Conclusion : This study showed that ECP hindered the process of rheumatoid arthritis and protected joints and cartilage.
Purpose: This study examined the effects of swimming exercise and Achyranthes Radix extracts on the inflammatory and behavioral responses in type II collagen-induced arthritic rats for 28 days. Methods: Sprague-Dawley rats were allocated randomly to one of the following four groups: only type II collageninduced (group Ⅰ), application of swimming exercise after type II collagen-induced (group II), application of Achyranthes Radix ointment after type II collagen-induced (group III), application of swimming exercise and Achyranthes Radix ointment after type II collagen-induced (group IV). Arthritis was established in SD rats by an intradermal injection of Chick type II collagen plus incomplete Freund's adjuvant at the base of the tail of the animals. The swimming exercise program consisted of a 25 min swimming session/day with a load corresponding to 5.5% of the weight bearing, three days/week for four weeks. The Achyranthes Radix ointment (0.1g) was applied twice a day for five days. The changes in behavior, H & E stain, and cyclooxygenase-2 (COX-2) level in the knee joint were assessed. Results: The gross and histological examination, after RA induction showed reddening, edema and erythema. The H & E stain revealed the destruction of articular cartilage, bony erosion and the infiltration of inflammatory cells after RA induction. The mechanical allodynia test results were significantly higher in group I than in groups II, III and IV (p<0.01). The immunohistochemistrical response of COX-2 in the knee joint showed that groups II, III, IV had a lower response effect than group I. Conclusion: Swimming exercise training and Achyranthes Radix ointment decreased the inflammatory responses and enhanced the behavioral responses in the arthritic rats.
Objectives : The purpose of this study is to observe the inhibitory effects of $Chrthami$ semen oil pharmacopuncture(CSOP) on CIA (collagen-induced arthritis) mice. Materials and Methods : Two types of experiments were conducted: $in$$vitro$ assay, inhibition of MIF mRNA and TNF-${\alpha}$ mRNA expressions in synovial membranes was observed, and $in$$vivo$ assay, $1{\mu}{\ell}/kg$ CSOP was injected every day to the left $Weizhong$ ($BL_{40}$) from day 3 to 21 after induction of CIA, and changes in paw edema, apical surface morphology, neovascularization in synovial membranes, fibrosis, pro-inflammatory cytokines production, Th-1 differentiation, and anti-inflammatory effect were investigated. Results : 1. In synoviocytes of the CIA mice treated with CSOP, MIF mRNA and TNF-${\alpha}$ mRNA expressions were down-regulated in a dose-dependent manner. 2. Paw edema of the CIA mice treated with CSOP was diminished. 3. Tissue injury in the synovial membranes, capillary distribution and fibrosis were reduced in CSOP-treated mice. 4. MIF, TNF-${\alpha}$, IL-6, MMP-9 expressions were repressed in CSOP-treated mice during the experiment to observe the inhibitory effect on cytokines production in early stage RA. 5. IL-12 and CD28 were reduced in CSOP-treated mice during the observation of inhibitory effect on Th 1 differentiation. 6. PPAR-${\gamma}$ was increased during the experiment to observe the anti-inflammatory effect of CSOP. Conclusions : The results may suggest that administration treatment using $Chrthami$ semen oil pharmacopuncture decreases the inflammatory response on an Animal Model with CIA.
Park, Min-Jung;Min, So-Youn;Park, Kyoung-Su;Cho, Mi-La;CHo, Young-Gyu;Min, Jun-Ki;Yoon, Chong-Hyeon;Park, Sung-Hwa;Kim, Ho-Youn
IMMUNE NETWORK
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v.5
no.4
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pp.221-231
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2005
Background: Immune regulatory dendritic cells (DCs) play an important role in maintaining self-tolerance. Recent evidences demonstrate that DCs expressing indoleamine 2,3-dioxygenase (IDO), which is involved in tryptophan catabolism, play an important role in immunoregulation and tolerance and induce T cell apoptosis. This study was devised to examine the role of IDO in the oral tolerance induction in collagen-induced arthritis (CIA) mouse model. Methods: Beginning 2 weeks before immunization, CII was fed six times to DBA/1 mice and the effect on arthritis was assessed. In tolerized mice, $CD11c^+$ DCs were isolated and stimulated with CII, IFN-${\gamma}$, and LPS with or without IDO inhibitor, 1-methyl-DL-tryptophan (1-MT) and IDO expression by $CD11c^+$ DCs was analyzed using FACS and RT-PCR. The expression of IDO, MHC II, CD80, and CD86 by $CD11c^+$ DCs were examined using confocal microscopy. Regulatory effect of $CD11c^+$ DCs on Ag-specific T cell proliferative response to CII was examined by mixed lymphocyte reaction (MLR) with or without 1-MT. Results: The proportion of IDO-expressing $CD11c^+$ DCs was slightly higher in tolerized mice than in CIA mice and significantly increased after stimulation with CII, IFN-${\gamma}$, and LPS in an IDO-dependent manner. On confocal microscopic examination, the expression of IDO was higher and those of MHC II and CD86 were lower in CD11c + DCs from tolerized mice compared to those from CIA mice. On MLR, $CD11c^+$ DCs from tolerized mice inhibited T cell proliferative response to CII in an IDO-dependent manner. Conclusion: Enhanced IDO expression by $CD11c^+$ DCs from tolerized mice may contribute to the regulation of proliferative response of CII-reactive T cells and could be involved in the induction of oral tolerance to CII.
Objectives The object of this study was to observe the favorable anti-arthritic effects of Samki-eum ($s\bar{a}nq\grave{i}-y\check{i}n$) on Freund's complete adjuvant (FCA)-induced arthritic Wistar rats. Methods Rheumatoid arthritis was induced by intradermal injection of FCA, and 300, 150 or 750 mg/kg of Samki-eum ($s\bar{a}nq\grave{i}-y\check{i}n$) were orally administered once a day for 14 days from 14 days after FCA treatments, and 15 mg/kg of dexamethasone was intraperitoneally administered as reference drug in this experiment. All rats were sacrificed at 14 days after continuous oral treatment of Samki-eum ($s\bar{a}nq\grave{i}-y\check{i}n$) or intraperitoneal administration of dexamethasone, and changes on the body weight, knee circumferences, gross arthritis score, inflammatory tissue prostaglandin (PG) $E_2$ levels were monitored with cartilage collagen components and glucosaminoglycans compositions - chondroitin sulphate, heparan sulphate and hyaluronic acid in the present study. Results As results of FCA treatment, classic rheumatoid arthritis featuring dramatical decreases on the body weights, cartilage collagen contents and bone glucosaminoglycans-chondroitin sulphate, heparan sulphate and hyaluronic acid contents, with increases on the knee circumferences, gross arthritis scores and inflammatory tissue $PGE_2$ levels. However, these changes from FCA-induced rheumatoid arthritis were clearly reduced by treatment of dexamethasone and both two different dosages of Samki-eum ($s\bar{a}nq\grave{i}-y\check{i}n$) 300 and 150 mg/kg in the present study. Although FCA-induced arthritis were more favorably inhibited by treatment of dexamethasone 15 mg/kg as compared with Samki-eum ($s\bar{a}nq\grave{i}-y\check{i}n$) 300 mg/kg, marked decreases of body weights were detected in dexamethasone 15 mg/kg treated rats, and Samki-eum ($s\bar{a}nq\grave{i}-y\check{i}n$) 300 mg/kg showed similar preserve effects on the cartilage glucosaminoglycan compositions in this study. Conclusions The results obtained in this study suggest that over 300 and 150 mg/kg of Samki-eum ($s\bar{a}nq\grave{i}-y\check{i}n$) showed favorable anti-arthritic effects on the FCA-induced arthritis mediated by suppression of $PGE_2$, a inflammatory mediator. However, detail mechanism studies should be conduced in future with the screening of the biological active compounds in this herb. Although overall anti-inflammatory effects Samki-eum ($s\bar{a}nq\grave{i}-y\check{i}n$) 300 mg/kg were lowered than those of dexamethasone 15 mg/kg treated rats, Samki-eum ($s\bar{a}nq\grave{i}-y\check{i}n$) 300 mg/kg treated rats showed similar preserve effects on the cartilage glucosaminoglycan compositions in this experiment.
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 $200{\mu}g/m{\ell}$.
Fangchinoline (FAN) is a bisbenzylisoquinoline alkaloid that is widely known for its anti-tumor properties. The goal of this study is to examine the effects of FAN on arthritis and the possible pathways it acts on. Human fibroblast-like synovial cells (FLS), carrageenan/kaolin arthritis rat model (C/K), and collagen-induced arthritis (CIA) mice model were used to establish the efficiency of FAN in arthritis. Human FLS cells were treated with FAN (1, 2.5, 5, 10 µM) 1 h before IL-1β (10 ng/mL) stimulation. Cell viability, reactive oxygen species measurement, and western blot analysis of inflammatory mediators and the MAPK and NF-κB pathways were performed. In the animal models, after induction of arthritis, the rodents were given 10 and 30 mg/kg of FAN orally 1 h before conducting behavioral experiments such as weight distribution ratio, knee thickness measurement, squeaking score, body weight measurement, paw volume measurement, and arthritis index measurement. Rodent knee joints were also analyzed histologically through H&E staining and safranin staining. FAN decreased the production of inflammatory cytokines and ROS in human FLS cells as well as the phosphorylation of the MAPK pathway and NF-κB pathway in human FLS cells. The behavioral parameters in the C/K rat model and CIA mouse model and inflammatory signs in the histological analysis were found to be ameliorated in FAN-treated groups. Cartilage degradation in CIA mice knee joints were shown to have been suppressed by FAN. These findings suggest that fangchinoline has the potential to be a therapeutic source for the treatment of rheumatoid arthritis.
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