BACKGROUND: Interleukin-17 (IL-17)-producing T helper (Th) 17 cells are considered as a new subset of cells critical to the development of rheumatoid arthritis (RA). We aimed to investigate the distribution of Th1 and Th17 cells and their association with disease activity, and determine the Th17-related cytokine levels in the peripheral blood of RA patients. METHODS: Peripheral blood mononuclear cells from 55 RA and 20 osteoarthritis (OA) patients were stimulated with mitogen, and the distributions of CD4+Interferon (INF)+IL-17- (Th1 cells) and CD4+INF-IL-17+ (Th17 cells) were examined by flow cytometry. Serum levels of IL-6, IL-17, IL-21, IL-23, and tumor necrosis factor (TNF)-alpha were measured by ELISA. Erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) were recorded. The 28-joint disease activity score (DAS28) was also assessed. RESULTS: The median percentage of Th17 cells was higher in RA patients than in OA patients (P=0.04), and in active than in inactive RA (P=0.03), whereas that of Th1 cells was similar in both groups. Similarly, the levels of IL-17, IL-21, and IL-23 were detected in a significantly higher proportion of RA patients than OA patients and the frequencies of detectable IL-6, IL-17, and IL-21 were higher in active RA than in inactive RA group. The percentage of Th17 cells positively correlated with the DAS28, ESR, and CRP levels. CONCLUSIONS: These observations suggest that Th17 cells and Th17-related cytokines play an important role in RA pathogenesis and that the level of Th17 cells in peripheral blood is associated with disease activity in RA.
Adult ; Aged ; Arthritis, Rheumatoid/blood/metabolism/*pathology ; Blood Sedimentation ; C-Reactive Protein/analysis ; Cytokines/blood ; Female ; Humans ; Male ; Middle Aged ; Osteoarthritis/blood/metabolism/pathology ; Severity of Illness Index ; Th1 Cells/cytology/immunology/metabolism ; Th17 Cells/*cytology/immunology/metabolism

OBJECTIVETo determine the levels of CC chemokine ligand 5 (CCL5) in serum and synovial fluid (SF) from patients with rheumatoid arthritis (RA) and their relations with disease activity and medication.

METHODSCCL5 in serum and SF was quantified by enzyme-linked immunosorbent assay (ELISA) in 28 RA patients and 21 osteoarthritis (OA) patients. In RA patients, the correlations of CCL5 levels in serum and SF with disease activity were analyzed. Meanwhile, the serum CCL5 levels among RA patients treated with disease-modifying antirheumatic drugs (DMARDs), Tripterygium Glucosides, and other Chinese herbs without disease-modifying effects were also compared.

RESULTSCCL5 levels in both serum and SF of RA patients were significantly higher than those of OA patients (P < 0.05). Moreover, the level of CCL5 was higher in SF than that in serum of RA patients (P < 0.01). Serum CCL5 level was correlated significantly with the number of swollen joints (r = 0.3329, P < 0.05), erythrocyte sedimentation rate (r = 0.4001, P < 0.05), and C reactive protein (r = 0.3735, P < 0.01). In addition, the level of CCL5 had a trend of lower in patients treated with DMARDs or Tripterygium Glucosides than those treated with other Chinese herbs, although the difference was not significant among those patients due to the small number of patients in each group.

CONCLUSIONSIn RA patients, the expression of CCL5 increases and correlates with some clinical and laboratory parameters of RA, which indicate that CCL5 plays an important role in RA and may serve as a useful marker of disease activity. DMARDs and Tripterygium Glucosides might exert their clinical effects through reducing CCL5 production in RA.

Adult ; Aged ; Arthritis, Rheumatoid ; blood ; drug therapy ; metabolism ; pathology ; Blood Sedimentation ; C-Reactive Protein ; metabolism ; Chemokine CCL5 ; analysis ; blood ; Female ; Humans ; Joints ; pathology ; Male ; Middle Aged ; Osteoarthritis ; blood ; metabolism ; Synovial Fluid ; metabolism ; Young Adult

OBJECTIVETo study reverse effect of the oxidative damage on cartilage cells of velvet antler polypeptides (VAPS), and to investigate the main mechanism of VAPS to protect cartilage cells through antioxidant.

METHODSFifteen Japanese white rabbits of 5-month-old were selected in this study. Animal model was established by method of Hulth osteoarthritis animal model. The anterior and posterior cruciate ligament and medial collateral ligament were cut off and medial meniscus were cut, articular cartilage cell cultured in vitro. Cells in the sham operation group was the normal control group, osteoarthritis cartilage cells in the model groups were added VAPS 6.25, 12.5, 25 microg/ml respectively. A group of animals were sacrificed every week form the ninth weeks(two months) and the cartilage cells were isolated and cultured. For 8 weeks,the reactive oxygen species level in chondrocytes were detected by DCFH-DA, the content of NO, SOD and GSH-Px in cell culture supernatant were detected by Griess method.

RESULTSDCFH-DA detection of intracellular reactive oxygen species was (5.46 +/- 0.46)in the control group, (12.08 +/- 0.74) in the model groups. The model group compared with the control group by t test with the P value less than < 0.001. DCFH-DA detection of intracellular reactive oxygen species was (9.81 +/- 0.59)in VAPS 6.25 microg/ ml group, (7.83 +/- 0.63) in the VAPS 12.5 microg/ml group, (6.89 +/- 0.71) in the VAPS 25 microg/ml group, as compared with model group there were statistically significant difference (P < 0.05). The content of NaNO2, SOD and GSH-Px in osteoarthritis model group was (5.60 +/- 0.45) microM, (38.56 +/- 12.53) U/ml and (151.90 +/- 25.60) U, as compared with control group there were statistically significant difference (P < 0.001, P < 0.05); The content of NaNO2 was (4.34 +/- 0.39), M in VAPS 6.25 microg/ml group, (3.67 +/- 0.36) microM in the VAPS 12.5 microg/ml group, (3.20 +/- 0.27) microM in the VAPS 25 microg/ml group, as compared with model group there were statistically significant difference (P < 0.01). The content of SOD was (49.91 +/- 5.77) U/ml in VAPS 6.25 microg/ml group, (54.05 +/- 5.27) U/ml in the VAPS 12.5 microg/ml group, (57.44 +/- 5.70) U/ml in the VAPS 25 microg/mL group, as compared with model group there was statistically significant (P < 0.05). The content of GSH-Px was (172.50 +/- 18.65) U in VAPS 6.25 microg/ml group, (202.10 +/- 21.60) U in the VAPS 12.5 microg/ml group, (315.80 +/- 10.50) U in the VAPS 25 microg/ml group, the VAPS 12.5 microg/mL group and VAPS 25 microg/ml group was compared with model group, there were statistically significant difference (P < 0.01).

CONCLUSIONThe VAPS have antioxidative damage effect of osteoarthritis cartilage cells within a certain range and dose-dependent manner. It may be the main mechanism for velvet antler polypeptides to treat osteoarthritis.

Animals ; Antlers ; chemistry ; Cartilage ; drug effects ; metabolism ; pathology ; Female ; Glutathione ; blood ; Male ; Nitric Oxide ; blood ; Osteoarthritis ; blood ; metabolism ; pathology ; Oxidative Stress ; drug effects ; Peptides ; pharmacology ; Rabbits ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; blood

OBJECTIVETo observe the metabolic, regulatory and anti-oxidative effects of modified Bushen Huoxue Decoction (BSHXD), a Chinese herbal medicine for kidney (Shen)-reinforcement and blood-activation, on an osteoarthritis (OA) rabbit model.

METHODSA rabbit model for knee joint OA was established by the classic Hulth's method. The OA model rabbits were randomized into 5 groups: the model control group, the positive control group treated with glucosamine sulfate, and the three BSHXD treated groups treated respectively with low, moderate, and high doses of BSHXD. In addition, a normal control group and a sham-operated group were set up. Experimental animals were sacrificed after a 7-week treatment, and pathological changes in cartilaginous tissue were estimated using the Mankin criteria. Hydroxyproline (Hyp) and malonaldehyde (MDA) contents in blood serum and urine, as well as superoxide dismutase (SOD) activity and nitric oxide (NO) content in blood serum and knee joint synovial homogenates were detected.

RESULTSMankin scoring showed insignificant statistical differences between the various treatment groups (P >0.05), but all were better than the model control group (P <0.05). Serum and urinary contents of Hyp and MDA as well as serum and synovial levels of NO were significantly lower, but the SOD activity in blood serum and synovial tissue was higher in the BSHXD treated groups than in the model group P <0.01); the effect of BSHXD was dose-dependent to some extent.

CONCLUSIONThe modified BSHXD shows an effect of improving cartilage metabolism in experimental rabbits with OA, and possesses osteo-chondric protective effects in antagonizing peroxidation injury.

Animals ; Antioxidants ; pharmacology ; therapeutic use ; Cartilage, Articular ; drug effects ; pathology ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hydroxyproline ; blood ; urine ; Male ; Malondialdehyde ; metabolism ; Nitric Oxide ; blood ; Osteoarthritis ; blood ; drug therapy ; metabolism ; pathology ; Rabbits ; Superoxide Dismutase ; blood ; Synovial Membrane ; drug effects ; enzymology ; pathology

The interleukin-33 (IL-33)/ST2 pathway has emerged as an intercellular signaling system that participates in antigen-allergen response, autoimmunity and fibrosis. It has been suggested that IL-33/ST2 signaling has been involved in the pathogenesis of rheumatoid arthritis (RA), because IL-33 and its receptor have been specifically mapped to RA synovium. The aim of this study was to determine the levels of IL-33 and sST2 in sera and synovial fluids in patients with RA. The serum level of IL-33 was significantly higher in patients with RA (294.9 +/- 464.0 pg/mL) than in healthy controls (96.0 +/- 236.9 pg/mL, P = 0.002). The synovial fluid level of IL-33 was significantly higher in RA patients than in osteoarthritis patients. The level of serum sST2 was higher in RA patients than in healthy controls (P = 0.042). A significant relationship was found between the levels of IL-33 and IL-1beta (r = 0.311, P = 0.005), and IL-33 and IL-6 (r = 0.264, P = 0.017) in 81 RA patients. The levels of IL-33, sST2 and C-reactive protein decreased after conventional disease-modifying antirheumatic drugs treatment in 10 patients with treatment-naive RA. Conclusively, IL-33 is involved in the pathogenesis of RA and may reflect the degree of inflammation in patients with RA.
Adult ; Aged ; Antirheumatic Agents/therapeutic use ; Arthritis, Rheumatoid/blood/drug therapy/*pathology ; C-Reactive Protein/analysis ; Female ; Humans ; Interleukin-1beta/analysis/blood ; Interleukin-6/analysis/blood ; Interleukins/*analysis/blood ; Male ; Middle Aged ; Osteoarthritis/blood/pathology ; Receptors, Cell Surface/*analysis/blood ; Synovial Fluid/metabolism

OBJECTIVETo investigate the effect of Ermiao Recipe (, EMR) with medicinal guide Angelicae Pubescentis Radix (APR) on the homing of bone marrow stem cells (BMSCs) to focal zone in osteoarthritis (OA) rats.

METHODSForty-eight Sprague-Dawley rats were randomly assigned to the sham-operated, model, EMR, and EMR plus APR groups (12 rats in each group). The OA rat model was induced by anterior cruciate ligament transection and medial meniscus resection. All rats were injected with recombinant human granulocyte colonystimulating factor [rhG-CSF, 30 μg/(kg·d) for continuous 7 days], and rats in the EMR and EMR plus APR groups were treated with EMR or EMR plus APR at 1.6 or 1.9 g/(kg·d) for 3 or 6 weeks, respectively. Cartilage histopathologic changes were observed by hematoxylin and eosin staining. Chondrocytes apoptosis and cartilage matrix components were tested by transferase-mediated deoxyuridine triphosphate-biotin nick end labeling assay and special staining. Interleukin-1β (IL-1 β), tumor necrosis factor α (TNF-α), bone morphogenetic protein 2 (BMP-2), and transforming growth factor beta-1 (TGF-β1) in serum were detected by enzyme-linked immunosorbent assay or radioimmunoassay assay. Matrix metalloproteinase (MMP)-13, tissue inhibitors of metalloproteinase (TIMP)-1, bromodeoxyuridine (BrdU), cluster of differentiation 34 (CD34), and stromal cell derived factor 1 (SDF-1) were measured by immunohistochemistry assay.

RESULTSEMR and EMR plus APR significantly inhibited articular cartilage damage and synovium inflammation in OA rats at 3 or 6 weeks of treatment, the most obvious changes in these parameters were found in the EMR plus APR group. At 6 weeks, compared with EMR treatment, EMR plus APR remarkably inhibited chondrocytes apoptosis and the release of IL-1β and TNF-α, obviously decreased MMP-13 expression, and significantly increased expressions of proteoglycan, collagen, type II collagen and TIMP-1, serum levels of BMP-2 and TGF-β1 as well as expressions of BrdU, CD34 and SDF-1 in cartilage articular (P<0.01 or P<0.05).

CONCLUSIONThe medicinal guide APR improved the therapeutic effects of EMR on OA rats by promoting directional homing of BMSCs to focal zone.

Animals ; Apoptosis ; drug effects ; Bone Marrow Cells ; drug effects ; Bone Morphogenetic Protein 2 ; blood ; Bromodeoxyuridine ; metabolism ; Cartilage, Articular ; drug effects ; enzymology ; pathology ; Chemokine CXCL12 ; metabolism ; Chondrocytes ; drug effects ; pathology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; pharmacology ; Humans ; Interleukin-1beta ; blood ; Knee Joint ; drug effects ; pathology ; Male ; Matrix Metalloproteinase 13 ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; blood ; Osteoarthritis ; blood ; drug therapy ; pathology ; Rats, Sprague-Dawley ; Synovial Membrane ; pathology ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism ; Transforming Growth Factor beta1 ; blood ; Tumor Necrosis Factor-alpha ; blood