Collagen-induced Arthritis Control Research Articles

Long non-coding RNA ENSMUST00000197208 promotes a shift in the Th17/Treg ratio via the P2X7R-NLRP3 inflammasome axis in collagen-induced arthritis.

Recently, long non‑coding RNAs (lncRNAs) have been implicated in several human diseases, including arthritis. However, the role of lncRNAs in regulating the Th17/Treg ratio during the progression of collagen-induced arthritis (CIA) is poorly understood. Therefore, the aim of this study was to determine the role of the lncRNA ENSMUST00000197208 and the P2X7R-NLRP3 inflammasome axis in changes in the Th17/Treg ratio in CIA. To achieve this, the distribution of T cell subgroups in the spleen cells of a CIA mouse model and control mice was examined. Additionally, we examined the expression profile of ENSMUST00000197208 in a CIA mouse model and healthy mice. The results showed that ENSMUST00000197208 expression was significantly upregulated in the CIA models compared with the control group. Additionally, the P2X7R-NLRP3 inflammasome axis participated in the pathogenesis of CIA and knockdown of ENSMUST00000197208 inhibited CD4+ T cell differentiation into Th17 cells. Compared with the control group, joint inflammation was less visible in NLRP3 knockout mice. Additionally, the P2X7R-NLRP3 inflammasomeaxis, which is downstream of ENSMUST00000197208, can be positively targeted and regulated by ENSMUST00000197208 through miR-107. Overall, the findings of this study showed that the "lncRNA ENSMUST00000197208-miR 107-P2X7R/NLRP3" axis plays an important role in CIA and knocking down ENSMUST00000197208 can efficiently inhibit Th17 differentiation bysuppressing the P2X7R-NLRP3 inflammasomeaxis. Therefore, targeting this axis may represent a novel strategy for arthritis treatment.

Salvianolate Ameliorates Osteopenia and Improves Bone Quality in Prednisone-Treated Rheumatoid Arthritis Rats by Regulating RANKL/RANK/OPG Signaling.

Rheumatoid arthritis (RA) is closely associated with periarticular osteopenia and leads to a high risk of generalized osteoporosis. Although glucocorticoid (GC) treatment ameliorates joint degradation and manages inflammation in RA, GC application may induce further bone quality deterioration in RA patients. Current treatments for RA lack relevant strategies for the prevention and treatment of osteopenia in RA. In this study, we aimed to investigate whether salvianolate treatment ameliorated osteopenia in prednisone-treated RA rats. Lewis rats with collagen-induced arthritis (CIA) were administered prednisone (PDN) or PDN plus salvianolate (PDN+Sal) treatment for 90 days. The effects of Sal were investigated in PDN-treated CIA rats. To further evaluate the effects of Sal under inflammatory conditions, we investigated the effects of Sal treatment on the TNF-α-induced inflammatory response in MC3T3-E1 osteoblasts. Bone histomorphometry, bone mineral density (BMD), bone biomechanical properties, micro-computed tomography (micro-CT), immunohistochemistry, RT-PCR and western blot analyses were performed to evaluate the effects of Sal. The results demonstrated that RA induced bone loss and bone quality deterioration, with high bone turnover in CIA rats. PDN+Sal treatment significantly increased BMD and trabecular/cortical bone mass, suppressed inflammation, and improved bone biomechanical properties compared to CIA control and PDN treatment. PDN+Sal treatment significantly suppressed bone resorption and the RANKL and RANKL/OPG ratios compared to PDN. PDN+Sal and PDN treatment significantly inhibited TNF-α by 82 and 83%, respectively, and both suppressed inflammation in CIA rats. However, there was no significant difference between PDN+Sal and PDN treatment alone in regard to bone formation parameters or the management of inflammation and arthropathy. Sal significantly increased Osterix, OPN, and Col1a1 while decreasing RANKL, TRAF6, and TRAIL gene in TNF-α-induced MC3T3-E1 osteoblasts. Sal significantly increased Osterix, OPN and RUNX2 while decreasing NF-κB, TRAF6 and IL-1β protein in TNF-α-induced MC3T3-E1 osteoblasts. The results suggested that salvianolate treatment ameliorated osteopenia and improved bone quality in prednisone-treated RA rats, and the potential mechanism may be related to the regulation of the RANKL/RANK/OPG signaling pathway, TRAIL-TRAF6-NFκB signal axis, and downregulation of inflammatory cytokines. Salvianolate could be used as a promising supplemental therapeutic strategy to ameliorate osteopenia and improve bone quality in GC-treated RA patients.

Therapeutic Effect of Matrine on Collagen-Induced Arthritis Rats and Its Regulatory Effect on RANKL and OPG Expression.

Objective To investigate the effect of matrine on rats with collagen-induced arthritis (CIA) and its regulatory effect on receptor activator of nuclear factor-κB ligand (RANKL) and osteoprotegerin (OPG) expression. Methods Wistar rats (n = 6) and CIA rats (n = 30) were randomly divided into six groups: healthy, CIA control, low/medium/high matrine (25, 50, or 100 mg/kg, once per day for six weeks), and methotrexate (MTX) (2 mg/kg, once per week for six weeks). The degree of joint damage was evaluated by X-ray and HE staining. Bone marrow suppression was assessed by routine blood analysis. In addition, the levels of serum RANKL and OPG in the rats were measured by ELISA. Results The level of joint swelling and degree of joint damage assessed by ankle swelling measurements, AI score, X-ray, and HE staining were alleviated in the CIA rats treated with MTX or different doses of matrine. Furthermore, no obvious inhibitory effect was observed on the bone marrow of the CIA rats, regardless of the dose of matrine or treatment with 2 mg/kg MTX (P > 0.05). The levels of OPG in serum and the ratio of OPG/RANKL were higher, and RANKL expression was lower in the low/medium/high matrine group compared with that of the CIA control group. The serum levels of OPG and OPG/RANKL ratio increased with the matrine dose, while the opposite was observed for RANKL expression. Conclusion Matrine treatment was associated with a lower degree of bone destruction, increased OPG expression and OPG/RANKL ratio, and decreased RANKL expression in CIA rats. Thus, matrine may represent a novel drug candidate for the treatment of RA.

Muscadine Grape and Wine Polyphenols Alleviated the Severity of Collagen Induced Arthritis in Mice

Abstract Objectives The aim of this study was to investigate the effect of concentrated muscadine grape polyphenols (MGP) and muscadine wine polyphenols (MWP) on the onset and progression of collagen induced arthritis (CIA) in mice. Methods MGP and MWP were concentrated using Amberlite XAD16N resin. Polyphenol composition was determined on HPLC. Male DBA/1 mice were divided into four treatment groups as (1) healthy control, (2) CIA control, (3) CIA + MWP, and (4) CIA + MGP. Arthritis was induced by intradermal injection of type II collagen on days 0 and day 21. Mice in groups 3 and 4 were gavaged with MWP and MGP using a dose of 400 mg/kg body weight for a total of 21 days. Severity of CIA was evaluated using clinical arthritic score and inflammation in the hind-paws. Plasma levels of cytokines, proteases, and anti-collagen antibody were measured using ELISA. Results MGP and MWP contained anthocyanin 3, 5-diglucosides, flavonols, and ellagic acid. Mice in the CIA control group had an onset of arthritis on day 18, which was delayed to day 22 and 24 by MWP and MGP, respectively (P < 0.05). Severity of arthritis was much lower in mice gavaged with muscadine polyphenols after the onset. On day 42, the average arthritic score of mice in the CIA control group progressed to 7.75 on a scale of 0–16, while MGP and MWP significantly reduced this score to 3.75 and 4.00, respectively (P < 0.01). In addition, MGP and MWP significantly reduced the plasma concentration of TNF-α, IL-6, anti-collagen antibodies, and matrix metalloproteinase-3 in CIA mice but not to the same level of healthy mice. The plasma concentration of IL-17 was drastically elevated in CIA mice, but it was not affected by MWP or MGP like other cytokines. This suggested that muscadine polyphenols had limited impact on the Th17 cells in the immune system. Mice gavaged with MPG and MWP had comparable plasma cytokine content, suggesting similar anti-inflammation activity. Conclusions Muscadine grape and wine polyphenols blunt the development of arthritis in mice by reducing the levels of key inflammatory cytokines, antibodies, and proteases, and may offer a promising dietary approach to manage arthritic symptoms. Funding Sources Florida Department of Agriculture and Consumer Service and Florida Viticulture Advisory Council.

Protective Effects of Undenatured Type II Collagen (UC‐II ® brand) in Animal Models of Rheumatoid Arthritis

Objective Undenatured Type II Collagen (Lonza's UC-II® branded ingredient, Lonza) shows an ability to reduce inflammation of rheumatoid arthritis (RA) and osteoarthritis through oral tolerance, but the mechanisms underlying this protective effect remain unclear. Our goal is to understand how changes in the gut microenvironment affect tolerogenic responses upon oral administration of UC-II® supplementation. Hypothesis We hypothesize that systemic inflammation during inflammatory arthritis regulate local tolerogenic responses and inflammation. Identification and further intervention of these pathways may be a promising way to improve the therapeutic activity of UC-II® ingredient. Methods Murine Collagen-Induced Arthritis (CIA) was employed as a model of human RA. Mice with CIA were treated with UC-II® supplementation or vehicle by oral gavage (0.66 mg/kg and 5.94 mg/kg for 8 weeks, starting 2 weeks before CIA induction). Incidence and clinical scores were monitored throughout the study period. Animals were culled at week 9, when blood, tissues (gut and lymph nodes) and paws were collected. Histopathological scores were carried out, including H&E staining of gut tissue and paws. Additionally, pro-inflammatory cytokine IL-17 and IgGs against collagen were evaluated by ELISA. Immune cell populations in mesenteric lymph nodes (MLNs) and draining lymph nodes (DLNs) were evaluated by flow cytometry. Finally, RT-PCR was carried out to evaluate expression of inflammatory genes. Results Mice treated with 5.94 mg/kg UC-II® supplementation showed a reduced disease incidence compared with CIA control mice, although clinical scores were not altered. However, histological analysis of the joints indicated that UC-II® administration reduced cartilage and bone damage. Interestingly, UC-II® reverted the reduction of villi length observed in the small intestine of CIA control animals. UC-II® treatment affected local and systemic immune responses, since numbers of CD4 T cells, CD8 T cells and B cells in joint draining and mesenteric lymph nodes were significantly modulated. Inflammatory responses were attenuated in UC-II® treated mice, evidenced by a reduction in serum antibodies against collagen and a strong reduction in IL-17 expression. Conclusions UC-II® supplementation protected against cartilage and bone damage during murine experimental arthritis. This could be possibly attributed to reduced expression of IL-17. Blocking of IL-17 pathway has been shown to have a positive effect on bone and cartilage damage in inflammatory arthritis. Our data suggest that oral administration of UC-II® supplementation could represent a novel approach to target this pathway in chronic inflammation. Further understanding of the molecular mechanisms triggered in the gut tissue during RA may pave the way for optimization of UC-II®.

Berberine Delays Onset of Collagen-Induced Arthritis through T Cell Suppression.

There is evidence that berberine (BBR), a clinically relevant plant compound, ameliorates clinically apparent collagen-induced arthritis (CIA) in vivo. However, to date, there are no studies involving the use of BBR which explore its prophylactic potential in this model of rheumatoid arthritis (RA). The aim of this study was to determine if prophylactic BBR use during the preclinical phase of collagen-induced arthritis would delay arthritic symptom onset, and to characterize the cellular mechanism underlying such an effect. DBA/1J mice were injected with an emulsion of bovine type II collagen (CII) and complete Freund’s adjuvant (day 0) and a booster injection of CII in incomplete Freund’s adjuvant (day 18) to induce arthritis. Mice were then given i.p. injections of 1 mg/kg/day of BBR or PBS (vehicle with 0.01% DMSO) from days 0 to 28, were left untreated (CIA control), or were in a non-arthritic control group (n = 15 per group). Incidence of arthritis in BBR-treated mice was 50%, compared to 90% in both the CIA and PBS controls. Populations of B and T cells from the spleens and draining lymph nodes of mice were examined on day 14 (n = 5 per group) and day 28 (n = 10 per group). BBR-treated mice had significantly reduced populations of CD4+Th and CD4+CXCR5+ Tfh cells, and an increased proportion of Foxp3+ Treg at days 14 and 28, as well as reduced expression of co-stimulatory molecules CD28 and CD154 at both endpoints. The effect seen on T cell populations and co-stimulatory molecule expression in BBR-treated mice was not mirrored in CD19+ B cells. Additionally, BBR-treated mice experienced reduced anti-CII IgG2a and anti-CII total IgG serum concentrations. These results indicate a potential role for BBR as a prophylactic supplement for RA, and that its effect may be mediated specifically through T cell suppression. However, the cellular effector involved raises concern for BBR prophylactic use in the context of vaccine efficacy and other primary adaptive immune responses.

Artocarpus tonkinensis Extract Inhibits LPS-Triggered Inflammation Markers and Suppresses RANKL-Induced Osteoclastogenesis in RAW264.7

Artocarpus tonkinensis (At) leaf decoction, a traditional remedy prepared in North Vietnam by the Hmong ethnic group, is a tea extract rich in bioactive compounds that may have therapeutic effects in arthritis and backache. Indeed, it has been demonstrated that At is able to inhibit Th17 lymphocytes development and to protect mice in an experimental model of collagen-induced arthritis. By resorting to macrophage in vitro models of inflammation and osteoclastogenesis, we showed that At extract significantly reduced nitric oxide synthase 2 (NOS2) activity and IL-6 production by RAW 264.7 murine cells. Moreover, At demonstrated an anti-osteoclastogenic effect, as revealed by complete inhibition of TRAP-positive osteoclast formation and decreased expression of key osteoclast-related genes. This At activity likely relies on the inhibition of RANK downstream signaling pathway, as the activation of non-receptor tyrosine kinase Src is reduced upon RANKL-At exposure. Protective effect of At against bone loss was also enlightened in vivo by collagen-induced arthritis (CIA) experiment demonstrating that, although paw edema was only weakly opposed by drinking At decoction, bone and cartilage were well preserved in CIA+At mice and joint tissue expressed decreased levels of osteoclast marker genes respect to CIA control group. Maesopsin 4-O-β-D-glucoside (i.e., TAT-2, one of the main decoction bioactive components) was capable to contrast NOS2 activity, IL-6 expression and osteoclast formation, too, albeit to a lesser extent when compared to At decoction. Overall, this study enlightens another At cell target, macrophages, beside Th17 lymphocytes, and suggests that the anti-arthritic beneficial effects of At decoction largely derives from its ability to counteract not only inflammation, but also osteoclastogenesis.

Bone marrow mesenchymal stem cells improve bone erosion in collagen-induced arthritis by inhibiting osteoclasia-related factors and differentiating into chondrocytes

BackgroundRheumatoid arthritis (RA) is characterized by joint inflammation and damage to the cartilage and bone in collagen-induced arthritis (CIA). Mesenchymal stem cells (MSCs) can improve articular symptoms and reduce bone erosion in CIA rats; however, the underlying mechanism remains unknown. This study aimed to investigate the mechanism underlying MSC-induced improvement of bone destruction in CIA.MethodsWistar rats were divided into a normal group, CIA control group, MTX intervention group, and BMSC intervention group, each comprising 8 rats. Serum RANKL, OPG, and CXCL10 levels of all groups were determined via flow cytometry after 42 days of interventions. RANKL, OPG, TRAF6, CXCL10, and CXCR3 were detected on the synovial membrane via immunohistochemistry, and their relative mRNA levels were determined via RT-PCR analysis. BMSCs were labeled with GFP and administered to CIA rats via the tail vein. At different time points, the distribution of implanted GFP-MSCs in synovial tissues was observed using a fluorescence microscope, and the potential of GFP-MSCs to differentiate into chondrocytes was assessed via immunofluorescence analysis.ResultsBMSC transplantation improved joint inflammation and inhibited bone destruction in CIA rats. BMSCs inhibited the expression of serum CXCL10 and CXCL10 and CXCR3 expression at the synovial membrane. Moreover, protein and mRNA expression analyses revealed that BMSCs potentially regulated RANKL/OPG expression levels in the serum and synovial tissue. Upon implantation into CIA rats, GFP-MSCs were traced in the joints. GFP-positive cells were observed in the cartilage tissue from day 11 and until 42 days after transplantation. Anti-type II collagen/GFP double-positive cells were observed in the articular cartilage (especially damaged cartilage) upon immunofluorescence staining of anti-type II collagen.ConclusionsBMSCs improve bone destruction in CIA by inhibiting the CXCL10/CXCR3 chemotactic axis, regulating the RANKL/OPG ratio, and directly differentiating into chondrocytes.

CXCR3 antagonist AMG487 suppresses rheumatoid arthritis pathogenesis and progression by shifting the Th17/Treg cell balance

Rheumatoid arthritis (RA) is an autoimmune disease that is characterized by uncontrolled joint inflammation and damage to bone and cartilage. Previous studies have shown that chemokine receptors have important roles in RA development, and that blocking these receptors effectively inhibits RA progression. Our study was undertaken to investigate the role of AMG487, a selective CXCR3 antagonist, in DBA/1J mice bearing collagen-induced arthritis (CIA). Following induction of CIA, animals were treated with 5 mg/kg AMG487 intraperitoneally every 48 h, starting from day 21 until day 41 and evaluated for clinical score, and histological hallmarks of arthritic inflammation. We further investigated the effect of AMG487 on Th1 (T-bet), Th17 (IL-17A, RORγt, STAT3), Th22 (IL-22), and T regulatory (Treg; Foxp3 and IL-10) cells in splenic CXCR3+ and CD4+ T cells using flow cytometry. We also assessed the effect of AMG487 on T-bet, RORγt, IL-17A, IL-22, Foxp3, and IL-10 at both mRNA and protein levels using RT-PCR and Western blot analyses of knee samples. The severity of clinical scores, and histological inflammatory damage decreased significantly in AMG487-treated compared with CIA control mice. Moreover, the percentage of Th1, Th17, and Th22 cells decreased significantly and that of Treg cells increased in AMG487-treated mice. We further observed that AMG487-treatment downregulated T-bet, IL-17A, RORγt, and IL-22, whereas it upregulated Foxp3 and IL-10 mRNA and protein levels. This study demonstrates the antiarthritic effects of AMG487 in CIA animal model and supports the development of CXCR3 antagonists as a novel strategy for the treatment of inflammatory and arthritic conditions.

Histological Evaluation of Lumbar Spine Changes in Rats with Collagen-induced Arthritis

To histologically evaluate lumbar involvement in rheumatoid arthritis (RA) by investigating rats with collagen-induced arthritis (CIA) and to assess the potential effects of RA on the discovertebral joints and facet joints. Seven-month-old female Sprague-Dawley rats were divided into groups with CIA and without CIA (control). All rats were sacrificed at 8 weeks after initial sensitization and the lumbar spine (L5/6) was harvested. Then the lumbar spine block specimens were stained with Villaneuva bone stain and sectioned in the midsagittal plane. The left facet joints were also sectioned in the midaxial plane. Specimens were studied under a microscope and infiltration of inflammatory cells was investigated. In the CIA group, lumbar lesions were observed in 13/18 rats (76%). Lymphocytes infiltrated into the anterior rim of the vertebral bodies only in 2 rats, while lymphocytes infiltrated the facet joints only in 4 rats. Both sites were involved in 7 rats. In addition, osteoclasts invaded the anterior rim of the vertebral bodies and formed cavities that also contained lymphocytes. Formation of pannus was seen in the facet joints in 11/18 rats. In CIA rats, infiltration of inflammatory cells into the anterior rim of the vertebral bodies alone or into the facet joints alone was demonstrated in 2 rats and 4 rats, respectively, while both sites were involved in 7 rats. Therefore, lesions at the anterior rim of the vertebral body did not arise secondary to facet joint involvement, but were caused by CIA along with synovial lesions of the facet joints.

Effect of allogenic mesenchymal stem cells transplantation on the expression of interleukin-22 and matrix metalloproteinase-3 in rats with collagen induced arthritis

Objective: To study the effect of allogeneic bone marrow mesenchymal stem cells transplantation on the expression of interleukin -22 (IL-22), matrix metalloproteinase -3 (MMP-3) in serum and synovial of rats with collagen induced arthritis. Methods: Type Ⅱ collagen were injected twice to establish the collagen induced arthritis (CIA) rat model. Forty-eight rats were randomly divided into 3 groups: control group, CIA control group, CIA experiment group. Rat bone marrow mesenchymal stem cells were cultured by bone marrow method combined with adherent culture method. After identify, the remaining cells were injected in the CIA experimental group. Enzyme linked immunosorbent assay (ELISA) and immunohistochemistry were used to detect the expression of IL-22 and MMP-3 in serum and anklebone joint's synovium of rats, respectively. Synovial cells were isolated and cultured, and were treated with different concentrations of IL-22. MMP-3 protein and mRNA were detected before and after stimulation by Western blot and real-time quantitative polymerase chain reaction (RT-qPCR). Results: After MSC transplantation, arthritis index, X-ray, HE staining of CIA rat showed that joint damage significantly reduced compared with the control group. The ELISA results showed that the expression of MMP-3 and IL-22 in CIA control group was higher than those in the control group (125.79±9.12 vs 102.00±7.63 ng/ml, P<0.05), (292.35±31.23 vs 257.27±13.99 ng/ml, P<0.05) and CIA experiment group (125.79±9.12 vs 97.94±9.50 ng/ml, P<0.05), (292.35±31.23 vs 262.16±22.02 ng/ml, P<0.05) with statistically significant difference (P<0.05). There was no significant difference between the control group and CIA experimental group. Immunohistochemical showed similar results with ELISA. Western blotting and RT-qPCR showed that MMP-3 protein and mRNA expression was increased after IL-22 stimulation in a dose-dependent manner. Conclusion: IL-22 and MMP-3 play an important role in the pathogenesis of rheumatoid arthritis. IL-22 could regulate the expression of MMP-3, and bone marrow mesenchymal stem cells could reduce the expression of MMP-3 in the treatment of rheumatoid arthritis by reducing the expression of IL-22.

Expression of peptidylarginine deiminase 4 and protein tyrosine phosphatase nonreceptor type 22 in the synovium of collagen-induced arthritis rats.

To study the expression level of peptidylarginine deiminase 4 (PADI4) and protein tyrosine phosphatase nonreceptor type 22 (PTPN22) in the synovium of rat model of collagen-induced arthritis, and to explore their possible therapeutic role in rheumatoid arthritis. Thirty-two female Wistar rats weighing 100±20 g were randomly assigned into 3-week collagen-induced arthritis (CIA) model group (n=8), 4-week CIA model group (n=8), 6-week CIA model group (n=8), and the control group (n=8). The body weight changes of each group were recorded. The expression levels of PADI4 and PTPN22 were detected and compared by the methods of immunohistochemical staining and Western blot. Arthritis of rat began to form 14 days after sensitization and the joint swelling reached peak at 28 days. The weights of the rats slowly grew both in CIA model groups and the control group. Immunohistochemical staining results showed that the positive expression of PADI4 and PTPN22 was mainly located in cartilage peripheral mononuclear cells, the cytoplasm of infiltrated cells, and bone marrow cavity. There were significant differences in the optical density of PADI4 and PTPN22 among CIA model groups and the control group (PADI4, 0.2898±0.012, 0.2982±0.022, 0.2974±0.031, 0.2530±0.013 in 3-week CIA model, 4-week CIA model, 6-week CIA model and control groups; PTPN22, 0.2723±0.004, 0.2781±0.010, 0.2767±0.008, 0.2422±0.019; all P <0.05). The expression bands of PADI4 were observed in Western blot 3 weeks after initial immunization, the thickest in the 4th week, and decreased in the 6th week. The expression bands of PTPN2 were observed at all the time points, with no obvious time-dependent trend. PADI4 and PTPN22 are obviously correlated with CIA in rat model. PADI4 is expressed at early stage of the disease, while the expression of PTPN22 sustains throughout the course.

Metabolomics analysis of collagen-induced arthritis in rats and interventional effects of oral tolerance

A serum metabolomics method based on rapid resolution liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (RRLC-Q-TOF-MS) was performed for a holistic evaluation of the metabolic changes of collagen-induced arthritis (CIA) in rats and to assess the interventional effects of type II collagen (CII) in this model. Partial least-squares-discriminant analysis (PLS-DA) was employed to study the metabolic profiling of CIA rats and control rats. Ten metabolites, namely, 12(S)-HHTrE, 12(S)-HEPE, PGE2, TXB2, 12(S)-HETE, LysoPE(16:0), PE(O-18:0/0:0), Lyso-PE(18:2), Lyso-PE(20:4), and Lyso-PC(22:5) were identified as differential metabolites associated with the pathogenesis of CIA. These results suggested that dysregulation of the arachidonic acid (AA) and phospholipid metabolic networks is involved in the pathomechanism of CIA. Differential metabolomics and histopathological analyses demonstrated that CII inhibits the progress of arthritis. Furthermore, the therapeutic effects of CII on CIA may involve regulation of the disordered AA and phospholipid metabolic networks. This metabolomics study provides new insights into the pathogenesis of arthritis and, furthermore, indicates the potential mechanism underlying the significantly increased prevalence of metabolic syndrome, defined as a clustering of cardiovascular disease (CVD) risk factors, in arthritis patients.

The Effects of Platycodin D, a Saponin Purified from Platycodi Radix, on Collagen‐Induced DBA/1J Mouse Rheumatoid Arthritis

The object of this study is to observe the effects of platycodin D, a saponin purified from Platycodi Radix, on mice collagen-induced arthritis (CIA). A daily dose of 200, 100, and 50 mg/kg platycodin D was administered orally to male DBA/1J mice for 40 days after initial collagen immunization. To ascertain the effects administering the collagen booster, CIA-related features (including body weight, poly-arthritis, knee and paw thickness, and paw weight increase) was measured from histopathological changes in the spleen, left popliteal lymph node, third digit, and the knee joint regions. CIA-related bone and cartilage damage improved significantly in the platycodin D-administered CIA mice. Additionally, myeloperoxidase (MPO) levels in the paw were reduced in platycodin D-treated CIA mice compared to CIA control groups. The level of malondialdehyde (MDA), an indicator of oxidative stress, decreased in a dose-dependent manner in the platycodin D group. Finally, the production of IL-6 and TNF-α, involved in rheumatoid arthritis pathogenesis, was suppressed by treatment with platycodin D. Taken together, these results suggest that platycodin D is a promising new effective antirheumatoid arthritis agent, exerting anti-inflammatory, antioxidative and immunomodulatory effects in CIA mice.

AB0118 Tissue balance (cartilage, synovium and connective tissue) in an animal model of rheumatoid arthritis

BackgroundSynovitis (synovial inflammation) is the main characteristics of rheumatoid arthritis (RA); a chronic inflammatory disease leading to progressive joint destruction. Synovitis is characterised by thickening of the synovial membrane due...

Changes of circulating sclerostin levels after different kinds of physical exercise with high impact on musculoskeletal system in healthy young females

Changes of circulating sclerostin levels after different kinds of physical exercise with high impact on musculoskeletal system in healthy young females

THE SERA FROM COLLAGEN-INDUCED ARTHRITIS MICE ACCELERATE MACROPHAGE-DERIVED FOAM CELLS FORMATION

ObjectivesRheumatoid arthritis (RA) is associated with excess of cardiovascular mortality and atherogenesis is accelerated in RA patients, but the exact mechanism remains unclear. Current findings support that the initiation and...

Establishment of Disease-syndrome Combination Rat Model of Rheumatoid Arthritis with Spleen Deficiency

Abstract This study was aimed at establishing a disease-syndrome combination rat model of rheumatoid arthritis (RA) with spleen deficiency. A group of 60 six-week-old SpragueDawley rats were randomly divided into normal control group and reserpine (RSP) group. Spleen deficiency model rat groups were made by an intraperitoneal injection of RSP. After 14 days, rats of RSP groups were subcutaneously injected with collagen type II protein to induce arthritis. The incidence of the model was observed, and the mechanism was analyzed by enzyme-linked immunosorbent assay (ELISA). The results showed that compared with the collagen-induced arthritis (CIA) group, the degree of joint swelling and joint damage was significantly increased in the RSP CIA model. The RSP CIA model rats were observed as exhibiting spleen deficiency syndrome that was similar to clinical symptoms, such as appearing apathetic, squinting, a sparse dry dull coat, a bowed back, a swollen ankle, fatigue, lazy movement, getting together, eating less, weight loss, and loose stools. Compared with the CIA control rats, spleen deficiency impaired the function of intestinal mucosa by disordering the intestinal villi, covering a large amount of goblet cells in the epithelium, and inducing exfoliation, degeneration, and necrosis fluff pieces. The content of anti-type II collagen antibody, IL-6, IL-10, and IFN-γ was increased. It was concluded that the rat model of rheumatoid arthritis with spleen deficiency syndrome by an intraperitoneal injection of reserpine has both obvious spleen deficiency syndrome and characteristics of RA. It can provide successful experiences for the rat model of rheumatoid arthritis with spleen deficiency syndrome. Spleen deficiency plays a vital role in RA.

The research of tolerance induced by oral collagen type II peptide-cholera toxin B subunit-liposome complex

Objective To investigate oral collagen type Ⅱ peptide-cholera toxin B subunit-liposome complex induce tolerance to collagen-induced arthritis (CIA).Methods DBA/1 mice were divided into four groups,group Ⅰ:normal control,group Ⅱ:CIA control,group Ⅲ:oral collagen type Ⅱ peptidecholera toxin B subunit-liposome complex,and group Ⅳ:for testing IgG2a (on day 14 after primary immunization).Arthritis scores and histopathologic assessment were analyzed.The levels of serum IgG2a were examined by ELISA.Results There was no arthritis development in group Ⅰ.The incidence of arthritis in group Ⅱ was higher than that in group Ⅲ ( 100％ vs 28.6％,P＜0.05).The arthritis score in group Ⅱ was higher than that in group Ⅲ (5.40 vs 0.4,3,P＜0.01).Histopathologic score was higher in group Ⅱ than that in group Ⅲ (16.00 vs 2.85,p＜0.05).Level of serum IgG2a of group Ⅰ was very 1ow(38 ng/ml).Mice of group Ⅱ produced significantly higher level of IgG2a than mice of group Ⅲ (3922 ng/ml vs 3219ng/ml,P＜0.05).IgG2a of group Ⅳ was 98 ng/ml which was significantly higher than that of group Ⅰ (P＜0.01 ).Conclusion Oral collagen type Ⅱ peptide-cholera toxin B subunit-liposome complex could inhibit CIA progression through immune tolerance. Key words: Collagen type Ⅱ peptide; Cholera toxin; Liposome; Immune tolerance; Arthritis

Establishment of a rat model of rheumatoid arthritis with kidney deficiency syndrome

To establish a rat model of rheumatoid arthritis (RA) with kidney deficiency syndrome. A total of 110 six-week-old specific pathogen-free male and female Sprague-Dawley rats were randomly divided into normal control group, sham-operated group, collagen-induced arthritis (CIA) control group, castration plus CIA group and hydroxyurea plus CIA group. Testiculus or ovary of rats in the castration plus CIA group was cut off, respectively. Rats in the hydroxyurea plus CIA group were given 375 mg/(kg·d) hydroxyurea by gavage administration for 17 d. Then rats in the CIA control group, castration plus CIA group and hydroxyurea plus CIA group were subcutaneously injected with mixture of type II collagen and incomplete Freund's adjuvant to induce rheumatoid arthritis. General state, arthritis index and joint swelling of the rats were observed to evaluate the onset of CIA. Contents of anti-type II collagen antibody, interleukin-6 (IL-6), IL-10, interferon-γ (IFN-γ) and corticosterone (CORT) in plasma were detected by enzyme-linked immunosorbent assay, and adrenal cyclic adenylic acid (cAMP) and cyclic guanylic acid (cGMP) levels were detected by radioimmunoassay. Compared with the CIA control group, the degrees of joint swelling and joint damage were significantly increased in the kidney-deficiency CIA rats (castration plus CIA group and hydroxyurea plus CIA group), with kidney deficiency syndrome similar to human clinical symptoms, such as depressed, bowed back, dullness, reduced diet and perianal contamination; the rats in those two groups were noted with a significantly decreased ratio of cAMP/cGMP; the content of CORT was increased in male rats while decreased in female rats, with an obvious increase in the content of anti-type II collagen antibody; the contents of IFN-γ, IL-6 and IL-10 were obviously increased in the castration plus CIA group. The rat model of RA with kidney deficiency syndrome has both obvious kidney deficiency syndrome and characteristics of RA and can reflect part of the patient's characteristics. However, castration is more suitable for inducing RA with kidney deficiency syndrome in rats.