Arthritis is a common disease in aged people, and is clinically divided into rheumatoid arthritis (RA) and osteoarthritis (OA). Although common symptoms such as pain are present, the underlying pathological mechanisms are slightly different. Therefore, the objectives of the present study were to compare joint damage induced by RA and OA by analyzing the major morphological and molecular differences, and to propose a suitable therapeutic intervention based on the pathophysiological conditions of bones and joints. For the RA animal model, 8-week-old DBA1/J mice were immunized with bovine type II collagen emulsified in complete Freund's adjuvant (CFA). Normal C57BL/6 mice (over 2 years of age) were used for OA. The clinical arthritis score was calculated using a subjective scoring system, and paw thicknesses were measured using calipers. The serum TNF ${\alpha}$ level was analyzed using an ELISA kit. Micro-CT was used to identify pathological characteristics and morphological changes. In collagen-induced RA mice, there were increased ankle joint volumes and clinical scores (p<0.01). The concentration of TNF ${\alpha}$ was significantly increased from 3 to 7 weeks after immunization. Micro-CT images showed trabecular bone destruction, pannus formation, and subchondral region destruction in RA mice. OA among aged mice showed narrowed joint spaces and breakdown of articular cartilage. This study suggests that a careful therapeutic intervention between RA and OA is required, and it should be based on morphological alteration of bone and joint.
In this study, we evaluated whether the oral administration of chondroitin from the cartilage of Raja kenojei is effective on the progression of rheumatoid arthritis (RA), using collagen-induced arthritic (CIA) mice. Arthritis development was delayed dose-dependently in the chondroitin-treated groups. The pre- and late-treated groups receiving 1,000 mg/kg of chondroitin had clinical scores that were reduced significantly by 56.9 (p<0.05) and 43.3% (p<0.05), respectively, compared to the vehicle-treated groups. Hematoxylin eosin staining and X-ray radiography showed that the chondroitins reduced the infiltration of inflammatory cells and prevented joint destruction of the knee and paw. Reverse transcription-polyerase chain reaction analysis revealed that chondroitin administration inhibited the expressions of tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), $interlukin-1{\beta}$ ($IL-1{\beta}$), and $interferon-{\gamma}$ ($IFN-{\gamma}$) in joints more than the administration of vehicle. Chondroitin treatment also decreased the production of rheumatoid factors (RF), IgG and IgM, in the serum of CIA mice. These results indicate that chrondroitin administration has a protective effect involving the inhibition of pro-inflammatory cytokine production in CIA mice.
Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by joint inflammation and progressive cartilage and bone erosion. Korean red ginseng (KRG) has been shown to have an anti-inflammatory effect by inhibiting the secretion of inflammatory cytokines like $TNF-{\alpha}$, IL-1, -6, and -8, and $IPN-{\gamma}$. In this study, whether KRG extract has an inhibitory effect on the collagen-inducible development of arthritis in DBA/1J mice was investigated. To induce arthritis, type II collagen emulsified in Complete Freund's Adjuvant was intradermally injected into the base of the tails of mice. Three weeks after the initial injection, a booster injection of type II collagen emulsified in Incomplete Freund's Adjuvant was administered. The oral administration of KRG extract for 8${\sim}$10 weeks at the dose of 300 mg/kg (three days a week) inhibited the development of arthritis in the experimental group, compared to the control group which was given saline. While the administration of KRG extract three times a week demonstrated both preventive and inhibitory effects, the administration of KRG extract once a week had little inhibitory effect. In other studies, the regimen of KRG administration has been shown to decrease the plasma level of inflammatory cytokines like IL-8 and TNF-${\alpha}$, but the plasma levels of these cytokines were not decreased in the present study. The results of the present study suggest that KRG has preventive and inhibitory effects on collagen-induced arthritis.
Objective & Methods : The purpose of this study is to observe the effects of Angelicae Pubescentis Radix herbal-acupuncture solution(APR-HAS) at Joksamni(ST36) on collagen IT induced arthritis in DBA/1J mice. The author performed several experimental items to analyze several cytokines and immune cells related with RA. Results : 1. In the APR-HA group, the incidence of arthritis and arthritis index were significantly decreased. 2. In APR-HA group, the levels of IL-6, $INF-{\gamma}$, $TNF-{\alpha}$, IgG, IgM, $IL-{\beta}$ and Anti-collagen II in serum of the CIA mouse were significantly decreased. 3. In APR-HA group, the level of $IFN-{\gamma}$, IL-4 in the CIA mouse spleen cell culture were significantly decreased. 4. In histology, the cartilage destruction and synovial cell proliferation were decreased in the APR-HA group, and the collagen fiber expressions in the APR-HA group were similar with that of the Normal group. 5. In the APR-HA group, CD3e+/CD19+ and CD4+/CD8+ were similarly maintained as Normal group in the CIA mouse lymph nodes, 6. In the APR-HA group, CD3e+/CD69+ was significantly decreased in the CIA mouse joint. 7. In the APR-HA group, CD11a+/CD19+ and CD11b+/Gr-l+ were significantly decreased in the CIA mouse lymph nodes 8. In the APR-HA group, CD4+/CD25+ was decreased in the CIA mouse spleen cell. 9. In the APR-HA group, CD4+/CD25+ was similarly maintained as Normal group in the CIA mouse lymph nodes.
Gwak Jung Mun;Oh Min Suck;Park Jong Ho;Kim Dong Hee
Journal of Physiology & Pathology in Korean Medicine
/
v.16
no.4
/
pp.810-817
/
2002
This study was carried out to know the effects of sopunghwalhyeoltang-gamibang(hereinafter referred to SPHHT) on the inhibition of arthritis induced by collagen on the mouse. Various experimental were performed in vivo (in DBA/1 J mice which are experimental model of arthritis induced by collagen) to analyse the immunomodulatory effects of SPHHT. The results were obtained as follows: 1. The cytotoxicity against mLFC was not measured in all concentration. 2. The arthritis index, incidence, hind paw edema, delayed-type hypersensitivity reaction, spleen weight were reduced in SPHHT treated group. 3. The expression of CD3 ε +/CD4+, CD3 ε +/CD8+ and CD19+ in peripheral blood mononuclear were reduced in SPHHT treated group. 4. The production of pro-inflammatory cytokines such as IL-6 and TNF-α were significantly reduced in SPHHT treated group. 5. The formation of new bones and synivium were stimulated in SPHHT treated group. Comparison of the results for this study showed that Sopung-hwalhyeoltang-gamibang had immunomodulatory effects. So we expect that Sopunghwalhyeoltang-gamibang should be used as a effective drugs for not only rheumatoid arthritis but also another auto-immune disease. Therefore we have to survey continuously in looking for the effective substance and mechanism in the future.
The conceptus which are resulted by mating between two genetically non-identical partners can be considered to be an allograft to the mother science which is not rejected by the mother's immunological attack. The present studies have been, therefore, attempted in order to elucidate the mechanism by which protection of the fete-placental allograft, between the C3H/HeJ female mouse and DBA/2 male mouse occurred. For this purpose, firstly systemic immunity was investigated by measuring T and B lymphocytes subsets. Natural killer cell activity in maternal splenic tissue and by observing the effects of pregnancy serums, progesterone and hCG on immune systems. Secondly, local immunity also investigated by measuring T lymphocytes subsets, natural killer cell activity in lymph nodes draining the uterus. The subsets of Thy-1.2$^+$ cells and L 3T4$^+$ cells decreased slightly while the subsets of Ly2$^+$ cell increased significantly compared with those of the control group beyond the mid-gestational stage. The subsets of B cell gradually in-creased from the mid-gestational stage untill delivery. The natural killer cell activity in the maternal splenic tissue significantly increased during the period of 5th to 8th day of gestation. The natural killer cell activity was significantly suppressed by the pregnancy serums and non-pregnant serums compared with those of serum-free group. The treatment of hCG significantly suppressed natural killer cell activity in the dose dependent manner (1 unit/ml-1000 unit/ml) while pro-gesterone increased the natural killer cell activity at phamarcological dose only. In the lymph nodes draining the uterus, the subsets of Thy-1.2$^+$ cells significantly increased during the period of implantation and L3T4$^+$ cell subsets slightly increased during the mid-gestational stage. The subsets of Ly2$^+$ cell increased significantly during the mid-gestational stage, but decreasing slightly be-fore delivery. The natural killer cell activity was significantly elevated after the implantation period in the lymph nodes draining the uterus. The natural killer cell activity of the lymph nodes draining the uterus was higher than those of splenic tissue during the same periods of gestation. It is therefore, concluded that during the pregnancy, the phenomena which the fete-placental allograft has not been rejected and rather protected from the maternal immunological attack might be due to local immune suppression in fete-maternal interface tissues rather than systemic immune suppression. And the subsets of Thy-1.2$^+$ cells and L3T4$^+$ cells mainly contribute to accepting allograft in early stage of pregnancy, while the subsets of Ly2$^+$ cell and the subsets of B cell increased significantly compared with those of the control group beyond the mid-gestational stage, so their role in systemic immunity and local immunity gradually increased from the mid-gestational stage until delivery.
Objective : The aim of this study is to observe the effect of Herbal-acupuncture(HA) with Luffae Fructus Retinervus Herbal-Acupuncture Solution(LFR-HAS) at ST36(Joksamni) on Collagen Ⅱ -induced arthritis(CIA) in mice. Methods : DBA1/J mice were immunized with bovine type Ⅱ collagen(CⅡ) on days 0 and 21 to induce an arthritis. The mice were divided into 5 groups. They were Normal group(wild type), Control group(CIA), Saline group(CIA +saline injection), Needle Prick group(CIA +single Prick with an injection needle) and LFR-HA group(CIA +LFR-HA treatment). The saline injection, needle prick and LFR-HA were made on the right ST36(Joksamni) of mice for 5 weeks, 3 times a week beginning 4 weeks after the booster immunization. Results : 1. The highest synovial rate of lung fibroblasts was measured in the 1% LFR-HAS. 2. TNF-${\alpha}$ expression of survival cells from CIA mouse joint was significantly reduced in the 1% LFR-HAS. 3. The incidence of arthritis and the spleen weight of CIA mouse were significantly reduced by the Luffae Fructus Retinervus Herbal-Acupuncture (LFR-HA) at ST36. 4. The concentrations of IL-6, INF-${\alpha}$, INF-${\alpha}$, IgG, IgM, and anti-collagen Ⅱ in the CIA mouse serum were significantly reduced by the LFR-HA at ST36. 5. The histological examination showed that, in the LFR-HA group, the cartilage destruction and the synoviocyte proliferation in the CIA mouse joint were not significant compared to the control group, and the collagen fiber was similarly expressed as the normal group. 6. In the LFR-HA group, the ratio of CD3e+ to CD19+ cell, and the ratio of CD4+ to CD8+ cell in the lymph node were similarly maintained as those of the normal group. 7. CD69+/CD3e+ and CD11a+/CD19+ cells in the CIA mouse lymph node were significantly reduced by the LFR-HA at ST36. 8. CD11b+/Gr-1+ cells in the CIA mouse joint were significantly reduced by the LFR-HA at ST36. Conclusions : These results indicate that Luffae Fructus Retinervus Herbal-Acupuncture (LFR-HA) at ST36 may regulate the immune system and have a therapeutic effect on Collagen-induced arthritis(CIA) in mice.
Kim, Il-Hyun;Lee, Ha-Il;Lee, Se-Won;Kwon, Young-Mi;Song, Yung-Sun
Journal of Korean Medicine Rehabilitation
/
v.25
no.1
/
pp.27-44
/
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.
Kim, Ji-Hoon;Shin, Mi-Jung;Cho, Hee-Jae;Lee, Sang-Won;Jeong, Jong-Moon
Korean Journal of Food Science and Technology
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v.33
no.6
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pp.656-663
/
2001
The most representative nitrosamine derived from nicotine, nitrosamine-4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone(NNK), has been reported to cause lung cancer in A/J mice. It has been also demonstrated that NNK-induced lung tumorigenesis involves $O^6-methylguanine(O^6MeG)$ formation, leading to $GC\;{\rightarrow}\;AT$ transitional mispairing during DNA replication. Our in vitro experiment, modified from the method of DBA assay, examined the ability of phyto-extract mixture to inhibit the metabolism of nicotine to nitrosamines. The production of nitromorpholine from morpholine was inhibited about 75% at the concentration of 20 mg/mL of phyto-extract mixture, which was lower than vitamine C and green tea powder. NNK, which is a pro-carcinogen in laboratory animals, is hydroxylated primarily in liver and lung by CYP 1A2, 2A6 and 3A4. A critical phase. of NNK activation is its change to an unstable metabolite methyl-diazohydroxide via CYP-mediated ${\alpha}-hydroxylation$; and then it provides a methyl group to the DNA to form DNA adducts which can easily induce mutations. $Aroclor^R$ 1254 was used to induce CYPs in the liver of a Sprague-Dawley rat. The ability of various test samples to inhibit CYPs that participate in NNK activation was evaluated, following the removal of the liver from the rat. Microsomal CYPlA2 catalyzing the conversion of NNK into strong carcinogenic chemicals was inhibited more efficiently by phyto-extract mixture than green tea powder. These results indicate that phyto-extract mixture can be used to reduce $O^6MeG$ DNA adducts for chemoprevention.
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.
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