Rheumatoid arthritis (RA) and osteoarthritis (OA), two common types of arthritis, affect the joints mainly by targeting the synovium and cartilage. Increasing evidence indicates that a significant network connects synovitis and cartilage destruction during the progression of arthritis. We recently demonstrated that hypoxia-inducible factor (HIF)-2alpha causes RA and OA by regulating the expression of catabolic factors in fibroblast-like synoviocytes (FLS) or chondrocytes. To address the reciprocal influences of HIF-2alpha on FLS and chondrocytes, we applied an in vitro co-culture system using a transwell apparatus. When co-cultured with HIF-2alpha-overexpressing chondrocytes, FLS exhibited increased expression of matrix metalloproteinases and inflammatory mediators, similar to the effects induced by tumor-necrosis factor (TNF)-alpha treatment of FLS. Moreover, chondrocytes co-cultured with HIF-2alpha-overexpressing FLS exhibited upregulation of Mmp3 and Mmp13, which is similar to the effects induced by interleukin (IL)-6 treatment of chondrocytes. We confirmed these differential HIF-2alpha-induced effects via distinct secretory mediators using Il6-knockout cells and a TNF-alpha-blocking antibody. The FLS-co-culture-induced gene expression changes in chondrocytes were significantly abrogated by IL-6 deficiency, whereas TNF-alpha neutralization blocked the alterations in gene expression associated with co-culture of FLS with chondrocytes. Our results further suggested that the observed changes might reflect the HIF-2alpha-induced upregulation of specific receptors for TNF-alpha (in FLS) and IL-6 (in chondrocytes). This study broadens our understanding of the possible regulatory mechanisms underlying the crosstalk between the synovium and cartilage in the presence of HIF-2alpha, and may suggest potential new anti-arthritis therapies.
Animals ; Arthritis/genetics/*immunology/pathology ; Arthritis, Rheumatoid/genetics/immunology/pathology ; Basic Helix-Loop-Helix Transcription Factors/genetics/*immunology ; Cells, Cultured ; Chondrocytes/immunology/metabolism/*pathology ; Coculture Techniques ; Fibroblasts/immunology/metabolism/*pathology ; Gene Expression Regulation ; Interleukin-6/genetics/*immunology ; Male ; Mice ; Mice, Inbred C57BL ; Osteoarthritis/genetics/immunology/pathology ; Synovial Membrane/immunology/metabolism/*pathology ; Tumor Necrosis Factor-alpha/genetics/*immunology ; Up-Regulation

Osteoarthritis (Osteoarthritis, OA) is a common clinical degenerative joint disease with increased incidence rate in recent years. Animal experiment is one of the important ways to explore pathogenesis and treatment of OA, while induced animal model is the most important part in animal experiment. Intra-articular injection of drugs is a classical method for establishing animal model of OA. Choose of animal should follows the principle of correlation, appropriateness and practicability, injections should perform in accordance with experimental purposes and subject, detections means and evaluation methods also should corresponding to experimental reality. The gold standard of OA animal model and intra-articular injections has not build, need further study.
Animals ; Cytokines ; analysis ; Disease Models, Animal ; Injections, Intra-Articular ; Mice ; Osteoarthritis ; diagnosis ; etiology ; immunology ; Rabbits ; Rats

INTRODUCTIONOsteoarthritis (OA) is a progressive degenerative disorder of the articular cartilage. Available diagnostic radiography has been poorly associated with the progress and severity of this clinical disease. As osteogenic protein-1 (OP-1) has been identified as a bone morphogenetic protein with a major role in cartilage repair, we aimed to evaluate its potential role in the diagnosis of OA.

METHODSThis was an experimental study conducted at the Department of Biochemistry, Sikkim Manipal Institute of Medical Sciences, India. Polyclonal antibodies (i.e. anti-OP-1[f]) were raised against OP-1 in mice, and subsequently used in a sandwich enzyme-linked immunosorbent assay (ELISA) to detect the presence of OP-1 in the synovial fluids of 75 osteoarthritic patients. For the purpose of correlation, the radiographic assessments of the knees of the 75 patients were graded using the Kellgren-Lawrence scoring system.

RESULTThe polyclonal antibody (i.e. anti-OP-1[f]) raised against OP-1 was able to detect the presence of OP-1 in the synovial fluids of all the osteoarthritic patients via sandwich ELISA. The level of the OP-1 was found to be much higher than the reference range and correlated positively with the severity of OA (r = 0.24; p = 0.04).

CONCLUSIONOur study shows that the polyclonal antibody, anti OP-1(f), could be used for the immunodiagnosis of osteoarthritis via sandwich ELISA.

Adult ; Aged ; Aged, 80 and over ; Animals ; Antibodies ; chemistry ; Bone Morphogenetic Protein 7 ; chemistry ; immunology ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Knee ; physiopathology ; Mice ; Middle Aged ; Osteoarthritis ; diagnosis ; immunology ; Synovial Fluid ; chemistry

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 explore the effects and mechanism of invigorating kidney and activating blood, invigorating kidney and expelling wind on hemorheology, IL-1β and TNF-α of SD rats with knee osteoarthritis, then definite the evolution of muscle certified turning into heumatism and compare the effect of Chinese herbal.

METHODSOne hundred and eighty SD rats with 3-month-old (each weight was 185 to 215 g) received intra-articular injection of papain solution for establishing knee OA models. All rats were randomly divided into activating blood group, preventing group, expelling wind group, invigorating kidney group, invigorating kidney and activating blood group and model group. Laboratory indexes were obtained at the 30th, 60th, 90th days after gastric perfusion, which including state of mind, activity, fur, weight, joint swelling, largely image, hemorheology, inflammation and HE pathological appearance.

RESULTSAfter operation, rats appeared blood stasis and swelling and difficulty crawling. There was significant difference of hemorheology in invigorating kidney and activating blood group the content of IL-1β and TNF-α was obviously lower than model group (P < 0.05 ). While the content of IL-1β and TNF-α on the early stage was obviously higher than late stage (P < 0.05).

CONCLUSIONKnee osteoarthritis mainly show synovial inflammation at the early stage, inflammation at early stage is more severe than late; invigorating kidney and activating blood decoction can inhibit the knee cartilage injury, improve blood circulation and prevent local inflammatory reaction. Activating blood decoction and invigorating kidney and activating blood Decoction have certain curative effect in early time, but the effects of invigorating kidney and activating blood Decoction is more effective than other on the late stage.

Animals ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Injections, Intra-Articular ; Interleukin-1beta ; immunology ; Kidney ; drug effects ; physiopathology ; Male ; Osteoarthritis, Knee ; drug therapy ; immunology ; physiopathology ; prevention & control ; Rats ; Tumor Necrosis Factors ; immunology

Osteoarthritis is a common joint disease, which seriously affects the patient's health and quality of life. It results in substantial social and economic costs. Etiology and pathogenesis of OA is still not completely clear, but people paid more attention on Cytokines, especially IL-1, which is considered as core factor in the development of OA. In recent years, many clinical trials considered IL-1 as a target treatment for OA. It provided a new treatment method. This article is to overview the mechanism of IL-1 in OA cartilage damage.
Animals ; Disease Progression ; Humans ; Interleukin-1 ; genetics ; immunology ; Osteoarthritis ; genetics ; immunology ; pathology

OBJECTIVETo explore the effects of Pilose antler polypeptide on apoptosis of chondrocyte and related cytokines in experimental knee osteoarthritis.

METHODSTotally 64 New Zealand White rabbits of 6 months old were randomly divided into 2 groups:normal group(n=8)and model group (n=56). Model group was surgically induced into knee osteoarthritis model by method of Hulth. After successful modeling,the rabbits of model group were further divided into 2 groups: Pilose antler polypeptide-treatment group (n=24) and control group (n=24). Pilose antler polypeptide-treatment group received 0.5 ml intraarticular injection of Pilose antler polypeptide dilution liquid once per 2 days for 30 days while control group received 0.5 ml intra-articular injection of physiological saline. On days 7, 15 and 30 after intervention, articular cartilage samples and synovial fluid were collected respectively. The morphological changes of articular cartilage under optical microscope and the structural change of chondrocyte were observed by transmission electron microscopy. The levels of interleukin-1beta and tumor necrosis factor-alpha in synovial fluid was detected by Enzyme-linked Immunosorbent Assay.

RESULTSAlong with the extending of time, articular cartilage degenerated gradually and chondrocytes apoptosis increased significantly. On days 7,15 and 30 after intervention, the chondrocyte apoptosis index of the Pilose antler polypeptide-treatment group were (20.30 +/- 1.23), (28.60 +/- 2.37), (37.10 +/- 1.82) and those of control group were (31.50 +/- 2.44), (34.40 +/- 1.77), (42.30 +/- 2.33). There were significant differences between them (P<0.05). At the same time, the chondrocyte apoptosis index of the Pilose antler polypeptide-treatment group were lower than those of control group,which had a statistical significance (P<0.05). On days 7,15 and 30 after intervention, the levels of interleukin-1beta in synovia fluid of Pilose antler polypeptide-treatment group were (15.81 +/- 1.26), (12.59 +/- 1.42), (9.57 +/- 0.92) microg/L and the level of tumor necrosis factor-alpha were (48.47 +/- 2.64), (43.46 +/- 1.33), (40.96 +/- 1.05) microg/L, with statistical differences(P<0.05). The levels of interleukin-1beta in synovia fluid of control group were (18.92 +/- 1.83), (20.25 +/- 2.76), (22.13 +/- 2.24) microg/L and the levels of tumor necrosis factor-alpha were (57.92 +/- 2.12), (60.25 +/- 1.48), (63.35 +/- 2.15) microg/L. At the same period,the levels of interleukin-1beta and tumor necrosis factor-alpha were lower than those of the control group,which had a statistical significance (P<0.05).

CONCLUSIONPilose antler polypeptide can inhibit chondrocytes apoptosis, decrease the levels of interleukin-1beta and tumor necrosis factor-alpha and delay the degeneration of articular cartilage to some extent.

Animals ; Antlers ; chemistry ; Apoptosis ; drug effects ; Chondrocytes ; drug effects ; pathology ; Female ; Interleukin-1beta ; analysis ; Male ; Osteoarthritis, Knee ; drug therapy ; immunology ; pathology ; Peptides ; pharmacology ; Rabbits ; Tumor Necrosis Factor-alpha ; analysis

Osteoarthritis (OA) is an age-related joint disease that is characterized by degeneration of articular cartilage and chronic pain. Oxidative stress is considered one of the pathophysiological factors in the progression of OA. We investigated the effects of grape seed proanthocyanidin extract (GSPE), which is an antioxidant, on monosodium iodoacetate (MIA)-induced arthritis of the knee joint of rat, which is an animal model of human OA. GSPE (100 mg/kg or 300 mg/kg) or saline was given orally three times per week for 4 weeks after the MIA injection. Pain was measured using the paw withdrawal latency (PWL), the paw withdrawal threshold (PWT) and the hind limb weight bearing ability. Joint damage was assessed using histological and microscopic analysis and microcomputerized tomography. Matrix metalloproteinase-13 (MMP13) and nitrotyrosine were detected using immunohistochemistry. Administration of GSPE to the MIA-treated rats significantly increased the PWL and PWT and this resulted in recovery of hind paw weight distribution (P < 0.05). GSPE reduced the loss of chondrocytes and proteoglycan, the production of MMP13, nitrotyrosine and IL-1beta and the formation of osteophytes, and it reduced the number of subchondral bone fractures in the MIA-treated rats. These results indicate that GSPE is antinociceptive and it is protective against joint damage in the MIA-treated rat model of OA. GSPE could open up novel avenues for the treatment of OA.
Analgesics/*administration & dosage ; Animals ; Antioxidants/*administration & dosage ; Bone Resorption ; Disease Models, Animal ; Gene Expression Regulation ; Humans ; Interleukin-1beta/genetics/metabolism ; Iodoacetates/administration & dosage ; Knee Joint/*drug effects/metabolism/pathology ; Male ; Matrix Metalloproteinase 13/genetics/metabolism ; Osteoarthritis/chemically induced/*drug therapy/physiopathology ; Pain ; Plant Extracts/administration & dosage ; Proanthocyanidins/*administration & dosage ; Rats ; Rats, Wistar ; Seeds ; Tomography, Emission-Computed ; Tyrosine/analogs & derivatives/metabolism ; Vitis/immunology

Osteoarthritis (OA) is an age-related joint disease that is characterized by degeneration of articular cartilage and chronic pain. Oxidative stress is considered one of the pathophysiological factors in the progression of OA. We investigated the effects of grape seed proanthocyanidin extract (GSPE), which is an antioxidant, on monosodium iodoacetate (MIA)-induced arthritis of the knee joint of rat, which is an animal model of human OA. GSPE (100 mg/kg or 300 mg/kg) or saline was given orally three times per week for 4 weeks after the MIA injection. Pain was measured using the paw withdrawal latency (PWL), the paw withdrawal threshold (PWT) and the hind limb weight bearing ability. Joint damage was assessed using histological and microscopic analysis and microcomputerized tomography. Matrix metalloproteinase-13 (MMP13) and nitrotyrosine were detected using immunohistochemistry. Administration of GSPE to the MIA-treated rats significantly increased the PWL and PWT and this resulted in recovery of hind paw weight distribution (P < 0.05). GSPE reduced the loss of chondrocytes and proteoglycan, the production of MMP13, nitrotyrosine and IL-1beta and the formation of osteophytes, and it reduced the number of subchondral bone fractures in the MIA-treated rats. These results indicate that GSPE is antinociceptive and it is protective against joint damage in the MIA-treated rat model of OA. GSPE could open up novel avenues for the treatment of OA.
Analgesics/*administration & dosage ; Animals ; Antioxidants/*administration & dosage ; Bone Resorption ; Disease Models, Animal ; Gene Expression Regulation ; Humans ; Interleukin-1beta/genetics/metabolism ; Iodoacetates/administration & dosage ; Knee Joint/*drug effects/metabolism/pathology ; Male ; Matrix Metalloproteinase 13/genetics/metabolism ; Osteoarthritis/chemically induced/*drug therapy/physiopathology ; Pain ; Plant Extracts/administration & dosage ; Proanthocyanidins/*administration & dosage ; Rats ; Rats, Wistar ; Seeds ; Tomography, Emission-Computed ; Tyrosine/analogs & derivatives/metabolism ; Vitis/immunology

OBJECTIVETo compare the levels of STAT4 tyrosine phosphorylation in peripheral T-lymphocytes induced by IL-12 in rheumatoid arthritis (RA) and osteoarthritis (OA).

METHODSFrom May 2007 to August 2009, peripheral blood mononuclear cells (PBMCs) were isolated from RA patients [RA group, all the cases were female, the age was from 28 to 55 years with an average of (45.0 +/- 13.0) years] and OA patients [OA group, all the cases also were female; the age was from 55 to 75 years with an average of (67.0 +/- 9.6) years]. The purity of T-lymphocytes from PBMCs was accredited by flow cytometry. The IL-12 of 50 ng/ml added in T-lymphocytes, the levels of STAT4 tyrosine phosphorylation were detected by western blot after different time intervals (0, 10, 30, 60 min).

RESULTSThe purity of T-lymphocytes were above 91% through diremption and depuration for peripheral blood monouclear cells. The levels of STAT4 tyrosine phosphorylation in T-lymphocytes from RA induced by IL-12 were higher than that from OA in the different times (10, 30, 60 min); after 30 min, its levels from RA and OA achieved to crest value.

CONCLUSIONSTAT4 in peripheral T-lymphocytes of rheumatoid arthritis was more easily to be activated than osteoarthritis.

Adult ; Aged ; Arthritis, Rheumatoid ; immunology ; Female ; Humans ; Interleukin-12 ; pharmacology ; Middle Aged ; Osteoarthritis ; immunology ; Phosphorylation ; Polymorphism, Single Nucleotide ; STAT4 Transcription Factor ; genetics ; metabolism ; T-Lymphocytes ; drug effects ; metabolism ; Tyrosine ; metabolism