Interleukin (IL)-27 is a novel cytokine of the IL-6/IL-12 family that has been reported to be involved in the pathogenesis of autoimmune diseases and has a pivotal role as both a pro- and anti-inflammatory cytokine. We investigated the in vivo effects of IL-27 on arthritis severity in a murine collagen-induced arthritis (CIA) model and its mechanism of action regarding control of regulatory T (Tregs) and IL-17-producing T helper 17 (Th17) cells. IL-27-Fc-treated CIA mice showed a lower severity of arthritis. IL-17 expression in the spleens was significantly decreased in IL-27-Fc-treated CIA mice compared with that in the CIA model. The Th17 population was decreased in the spleens of IL-27-Fc-treated CIA mice, whereas the CD4+CD25+Foxp3+ Treg population increased. In vitro studies revealed that IL-27 inhibited IL-17 production in murine CD4+ T cells, and the effect was associated with retinoic acid-related orphan receptor gammaT and signal transducer and activator of transcription 3 inhibition. In contrast, fluorescein isothiocyanate-labeled forkhead box P3 (Foxp3) and IL-10 were profoundly augmented by IL-27 treatment. Regarding the suppressive capacity of Treg cells, the proportions of CTLA-4+ (cytotoxic T-lymphocyte antigen 4), PD-1+ (programmed cell death protein 1) and GITR+ (glucocorticoid-induced tumor necrosis factor receptor) Tregs increased in the spleens of IL-27-Fc-treated CIA mice. Furthermore, in vitro differentiated Treg cells with IL-27 exerted a more suppressive capacity on T-cell proliferation. We found that IL-27 acts as a reciprocal regulator of the Th17 and Treg populations in CD4+ cells isolated from healthy human peripheral blood mononuclear cells (PBMCs), as well as from humans with rheumatoid arthritis (RA) PBMCs. Our study suggests that IL-27 has the potential to ameliorate overwhelming inflammation in patients with RA through a reciprocal regulation of Th17 and Treg cells.
Animals ; Arthritis, Experimental/*drug therapy/immunology ; Cells, Cultured ; Humans ; Interleukins/immunology/*therapeutic use ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Inbred DBA ; T-Lymphocytes, Regulatory/*immunology ; Th17 Cells/*immunology

OBJECTIVETo compare the effects of Atractylodes lancea from different producing area on collagen-induced arthritis (CIA) in rats.

METHODWistar rats were induced by type II bovine collagen. CIA rats were treated daily with oral administration of A. lancea from the geo-authentic and non-authentic producing area of Maoshan, Jiangsu province, and non-geo-authentic and non-authentic producing areas of Yingshan, Hubei province and Huayin, Shanxi province from day 7 after the day of the first immunization to day 35. Clinical symptoms as well as clinical scores and incidence were observed. All rats were sacrificed on day 35 after immunization to observe histopathologic and radiologic changes. Antibody to type II collagen in sera was measured by enzyme-linked immunosorbent assay (ELISA), the levels of pro-inflammatory cytokines tumor necrosis factor alpha (TNF-alpha), interleukin-1beta (IL-1beta), and interleukin-6 (IL-6) in sera and article-homogenated supernants by radiommunoassay, and inflammatory mediator of PGE2 in sera using ELISA.

RESULTA. lancea from Jiangsu province can ameliorate clinical symptom, reduce arthritis index and arthropathy of inflammatory joints, inhibit the production of IgG and IgM in sera, directly suppress the production of exogenous and endogenous cytokines of IL-1beta, TNFalpha and IL-6 and PGE2. A. lances from Hubei and Shanxi provinces can inhibit the production of IgM in sera, and A. lanceas from Hubei province also depress the production of IL-1beta in sera and IL-6 in supernants.

CONCLUSIONA. lancea from Jiangsu province is effective in CIA rats through inhibiting the production of pro-inflammatory cytokines and the inflammatory mediators.

Animals ; Arthritis, Experimental ; drug therapy ; immunology ; Atractylodes ; Cytokines ; blood ; Dinoprostone ; blood ; Male ; Phytotherapy ; Plant Extracts ; therapeutic use ; Rats ; Rats, Wistar

OBJECTIVETo observe the therapeutic effect and mechanism of nux vomica total alkali gel (NVTAG) on adjuvants arthritis (AA) rats.

METHODSD rats were randomly divided into nine groups: the normal group, the AA model group, NVTAG high, middle and low-dose (25, 12.5, 6.25 mg x kg(-1)) groups and the Votalin control (diclofenac diethylamine emulgel, 50 mg x kg(-1)) group. Except for the normal group, the remaining groups were transcutaneously administered with 0.1 mL freund's adjuvant complete (FCA) for inflammation in left rear feet and then evenly treated with medicine and packed with oilpapers. The foot volume method was adopted to determine foot swelling degree, with pain scoring and polyarthritis scoring. HE staining was used to observe arthro-pathologic injury. The content of prostaglandin E2 (PGE2), interleukin-1 (IL-1), IL-6, tumor necrosis factor (TNF-alpha) and vascular epidermal growth factor (VEGF) in synovium homogenates were measured by enzyme-linked immuno-absorbent assay (ELISA) respectively.

RESULTCompared with the model group, NVTAG and control gel can obviously reduce the foot swelling degree, polyarthritis indicators and relieve arthro-pathologic injury in AA rats (17-21 d). The levels of IL-1, PGE2, IL-6, VEGF and TNF-alpha in synovial homogenates of AA rats were also reduced by NVTAG significantly.

CONCLUSIONNVTAG shows an antergic effect on AA progress in rats, which is closely related to inhibition of development of inflammatory mediator.

Alkalies ; Animals ; Arthritis, Experimental ; drug therapy ; immunology ; pathology ; Cytokines ; analysis ; Gels ; Male ; Phytotherapy ; Rats ; Rats, Wistar ; Strychnos nux-vomica

OBJECTIVETo observe the effects of Tripterygium polyglycosides (TWP) on mucous immune function in rat models of arthritis induced respectively with collagen-II induced arthritis (CIA) & adjuvant arthritis (AA).

METHODSCIA and AA model rats were induced by immunization with collagen II emulsified with complete Freund's adjuvant and complete Freund's adjuvant respectively and treated with TWP. Rats' mucus, systemic immunological indexes (peripheral subsets of T cells), local inflammatory factors (IL-6, TNF-alpha, COX-2,and NF-kappaB, etc. ) were observed.

RESULTSIn CIA model group, CD4+ in Peyer's Patch (PP), peripheral CD4+ and CD8+ positive T cells all raised, while in the AA model group, CD4+ lowered and CD8+ raised on PP, with both subsets increased. Effects of TWP on T lymphocyte subsets in PP and blood of the two models were different. High leveled IL-6, TNF-alpha, COX-2 and NF-kappaB expression could be seen in both model groups, and these inflammatory media could be inhibited by TWP.

CONCLUSIONThere exist similarities and differences between the two models in aspects of mucus immune response and effect of TWP on them.

Animals ; Arthritis, Experimental ; chemically induced ; drug therapy ; immunology ; Collagen Type II ; Freund's Adjuvant ; Glycosides ; pharmacology ; Male ; Mucous Membrane ; immunology ; Rats ; Rats, Sprague-Dawley ; Tripterygium ; chemistry

OBJECTIVETo study the effect of Yangqixue Qufengshi Recipe (YQXQFS) on rheumatoid arthritis (RA) model mice under different genetic backgrounds.

METHODSCollagen Induced Arthritis (CIA) were established on HLA-DR4 transgenic (TG) mice and non-transgenic (NTG) mice, which partly were raised with YQXQFS, and the onset day of CIA, the level of type II collagen (CII)-reactive antibodies and the pathological scores of CIA were assessed.

RESULTSUnder HLA-DR4 TG background (compared with NTG mice), the earlier onset day of CIA (11.22 +/- 3.35 days vs 16.56 +/- 4.75 days, P < 0.05) and higher level of CII-reactive antibodies (0.2274 +/- 0.1390 microg/ml vs 0.1101 +/- 0.0560 microg/ml, P < 0.05) were observed, but the pathological scores of CIA remained unchanged. YQXQFS could not influence the onset day of CIA and the level of CII-reactive antibodies, but had a certain effect on the total pathological scores (6.56 +/- 3.43 scores vs 11.11 +/- 5.64 scores) and bone erosion (0.22 +/- 0.44 scores vs 1.67 +/- 1.50 scores) of CIA on NTG mice (P < 0.05), NTG YQXQFS group compared with NTG experimental group.

CONCLUSIONYQXQFS had a certain effect on RA model, but had no significant effect on HLA-DR4 related CIA.

Animals ; Antibodies ; blood ; Antirheumatic Agents ; therapeutic use ; Arthritis, Experimental ; drug therapy ; Arthritis, Rheumatoid ; drug therapy ; immunology ; pathology ; Collagen Type II ; immunology ; Drugs, Chinese Herbal ; therapeutic use ; HLA-DR4 Antigen ; genetics ; Mice ; Mice, Inbred Strains ; Mice, Knockout

OBJECTIVETo study the modulation effect of pro- and anti-inflammatory cytokines following long term use of water soluble ethanol extracts from different organs of Nyctanthes arbortristis (NAT) in mouse model of arthritis.

METHODSArthritis was induced in mice by two injections of Freund's complete adjuvant on days 0 and 12 in the sub-planter surface of the right hind paw.

RESULTSInjection of adjuvant resulted in a maximum primary edema of the footpad with erythema, and edema and distortion of joints of the right hind paw after 24-48 hours. Second injection of FCA led to the formation of secondary swellings persisting more than four weeks that spread onto the other hind limb but to a lesser extent. Histological analysis of the ankle on day 47 showed marked evidence of cartilage destruction in association with pannus formation and moderate bone resorption. Proinflammatory cytokine levels in the inflamed joint homogenate were elevated on days 2, 14, and 47. Oral administration of leaf and fruit extracts in arthritic mice reduced joint homogenate levels of tumor necrosis factor-alpha, interleukin-1beta, and interleukin-6 on days 2, 14, and 47 in comparison to untreated arthritic mice. Interleukin-10 level was elevated in the inflamed joint on days 2, 14, and 47 in comparisons to untreated arthritic mice.

CONCLUSIONEvidence of lesser inflammation of the footpad and joint and associated histological observation support the therapeutic benefit of leaf and fruit extracts from Nyctanthes arbortristis. This study helps in understanding the mechanism of anti-inflammatory action of Nyctanthes arbortristis in the light of pro- and anti-inflammatory cytokine balance.

Animals ; Arthritis, Experimental ; drug therapy ; immunology ; pathology ; Cytokines ; metabolism ; Disease Models, Animal ; Female ; Mice ; Mice, Inbred BALB C ; Oleaceae ; chemistry ; Phytotherapy ; Plant Extracts ; isolation & purification ; therapeutic use

OBJECTIVETo study pharmacologic actions for rheumatoid arthritis treatment and probe into the effects and mechanisms of Simiao pill on adjuvant arthritis (AA) rats.

METHODWistar rats, male, were randomly divided into the normal comparison group, model group, the glucosida Tripterygii TOTA group (9.45 mg x kg(-1)), the Simiao pill low dose group (1 080 mg x kg(-1)), the Simiao pill middle dose group (2 160 mg x kg(-1)) and the Simiao pill high dose group (4 320 mg x kg(-1)). Except the normal comparison group, rats in the other groups were injected Freund's complete adjuvant reagent into the rats' right etapedes to establish the AA model. Drugs were given by intragastric administration. Then paw swelling, SOD in blood serum, the spleen and and thoracic gland index, histopathology change of malleolus joint were observed. IL-1beta, IL-6 and TNF-alpha mRNA were observed by semi-quantitative reverse transcription and polymerase chain reaction(sqRT-PCR).

RESULTSimiao pill could inhibit the swelling of the rats' inflamed metapedes. Compared with the model group, each group of Simiao pill could increase SOD, decrease spleen index, advance the thoracic gland index and decrease IL-1beta, IL-6 and TNF-alpha mRNA.

CONCLUSIONSimiao pill could threat AA rats and inhibit the inflammation of synovial membrane obviously. It also could relieve the degree of paw swelling, and therefore provide clinical theatment basis of RA.

Animals ; Arthritis, Experimental ; drug therapy ; genetics ; immunology ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Gene Expression ; drug effects ; Humans ; Interleukin-6 ; genetics ; immunology ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; genetics ; immunology

OBJECTIVETo investigate the effect of the total flavonoids of orange peel (TFO) against adjuvant arthritis (AA) and the underlying mechanism.

METHODAA model was induced in male Wistar (correction of SD) rats by immunization with Freund's complete adjuvant Pad thickness was assayed by caliper. Pathological impairment of ankle joint was analysised by hematoxylin and eosin (H&E) staining. Levels of tumor necrosis factor (TNF)-alpha, Interleukin (IL)-1beta and prostaglandin (PG) E2 in serum was detected by radioimmunoassay method. Cyclooxygenase (COX)-2 expression in synovium tissues was measured by Western blot assay.

RESULTThe 75 mg x kg(-1) and 150 mg x kg(-1) TFO treatment obviously decreased the pad thickness and improve the pathological impairment of ankle joint of AA rats. In addition, abnormal elevation of TNF-alpha, IL-1beta and PGE2 in serum and COX-2 expression in synovium tissues of AA rats were markedly repressed by TFO treatment.

CONCLUSIONTFO can inhibit the development of AA in rats, and the mechanism were likely due to depressing inflammatory mediators production.

Animals ; Arthritis, Experimental ; chemically induced ; drug therapy ; immunology ; Citrus sinensis ; chemistry ; Disease Models, Animal ; Flavonoids ; administration & dosage ; Freund's Adjuvant ; adverse effects ; Humans ; Interleukin-1 ; immunology ; Male ; Plant Extracts ; administration & dosage ; Random Allocation ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; immunology

BACKGROUNDCryptotanshinone (CT) is the major active constituent of Salvia miltiorrhiza Bunge. The present study was carried out to investigate the effects of CT on rats with adjuvant arthritis (AA).

METHODSAA was induced by the metatarsal footpad injection with complete Freund's adjuvant in male Sprague-Dawley rats. The secondary inflammatory reaction was evaluated by hind paw swelling and the polyarthritis index. Activity of interleukin-1 (IL-1) was detected by the concanavalin A-induced thymocytes proliferation assay. The lymphocytes proliferation and IL-2 production were assayed by 3-(4,5-2dimethylthiazal-2yl)2,5-diphenyltetrazoliumbromide (MTT) and activated mouse splenocytes proliferation, respectively.

RESULTSIntragastric administration of CT (50 and 100 mg/kg) significantly decreased secondary inflammatory reactions and increased the spleen and thymus index. There was a marked immunologic and inflammatory response in the AA model, which was accompanied by the decrease of thymocyte proliferation and IL-2 production as well as the increase of IL-1 production. CT apparently enhanced thymocyte proliferation and decreased IL-1 production in AA rats.

CONCLUSIONThese results indicate that CT may exert its anti-inflammatory and immunoregulatory effects through inhibiting lymphocyte proliferation and production of pro-inflammatory mediators.

Animals ; Arthritis, Experimental ; drug therapy ; immunology ; metabolism ; Cell Proliferation ; drug effects ; Interleukin-1 ; metabolism ; Interleukin-2 ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Phenanthrenes ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Thymocytes ; cytology ; drug effects ; metabolism

OBJECTIVETo determine the effective dosage and formulation of agkistrodon in collagen-induced arthritis (CIA) rats.

METHODSCIA was induced by injection of collagen in complete/incomplete Freund's adjuvant. Agkistrodon decoction, agkistrodon powder, and agkistrodon wine were administered daily starting from the onset of arthritis. Paw swelling degree was measured by using a volume-measuring instrument every 7 days after primary immunization. Arthritis index was measured and calculated using the "five scoring method" every 7 days. The levels of serum interleukin-1ß (IL-1ß) and type II collagen IgG antibodies were detected by enzyme-linked immunosorbent assay. Finally, all ankles were removed, and X-ray radiography was performed with In-vivo Imaging System FX. Samples were counterstained with hematoxylin and eosin for analysis.

RESULTSAmong the various dosage formulations of agkistrodon, high-dose powder, which was equivalent to an amount of 6 g/day in adults, showed better effects on the inhibition of joint swelling and reduction of arthritis index score. The relatively low levels of serum IL-1 and anti-type II collagen IgG antibodies, as well as the X-ray radiography and pathology results, further proved the superiority of the high-dose powder over the other formulations. The effect of decoction on inhibiting joint swelling was inversely proportional to the dosage. Other effects, such as reduction of arthritis index score and the levels of serum IL-1 and anti-type II collagen IgG antibodies, were directly proportional to the dosage. While the use of large dose agkistrodon wine led to negative effects.

CONCLUSIONThese data highlight the potential function of high-dose agkistrodon powder, which was equivalent to an amount of 6 g/day in adults. The powder can quickly relieve the symptoms of rheumatoid arthritis and prevent aggravation of disease, especially during the early period.

Agkistrodon ; metabolism ; Animals ; Antibodies ; blood ; Arthritis, Experimental ; blood ; chemically induced ; drug therapy ; Collagen Type II ; immunology ; Dosage Forms ; Dose-Response Relationship, Drug ; Enzyme-Linked Immunosorbent Assay ; Extremities ; diagnostic imaging ; pathology ; Female ; Interleukin-1beta ; blood ; Medicine, Chinese Traditional ; Rats, Wistar