Effects on Pycnogenol attenuating the inflammatory on joint inflammation induced by urate crystals and interleukin-1 beta
| Autor: | Peng, Yi-Jen, 彭奕仁 |
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| Rok vydání: | 2012 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 100 Objective Acute gouty arthritis results from monosodium urate (MSU) crystal deposition in joint tissues and mostly tophi deposit in the infrapatellar fat pad of knee. Deposited MSU crystals induce an acute inflammatory response which leads to damage of joint tissue. Interleukin-1β is a potent pro-inflammatory cytokine and plays a pivot role in osteoarthritis, rheumatoid arthritis and gouty arthritis. Pycnogenol (PYC), an extract from the bark of Pinus maritime, has documented anti-inflammatory and antioxidant properties. The present study aimed to investigate whether PYC had protective effects on IL-1β and MSU-induced inflammatory and nitrosative stress in joint tissues both in vitro and in vivo. Materials and methods Cultures of primary human articular chondrocytes, synovial fibroblasts and infrapatellar fat pad were obtained and generated from the knee arthroplasty specimens of osteoarthritic patient. Real-time polymerase chain reaction was used to assess gene regulation. Expressions of inducible protein and signaling protein were assessed by western blot and immunofluorescent stain. The secretions of IL-8, nitric oxide and MMP-9 were assayer by enzyme linked immunosorbent assay (ELISA), Griess reaction and gelatin zymography, respectively. Results MSU crystals upregulated cyclooxygenase 2 (COX-2), interleukin 8 (IL-8) and inducible nitric oxide synthase (iNOS) gene expression in human articular chondrocytes, but only COX-2 and IL-8 in synovial fibroblasts. PYC inhibited the up-regulation of COX-2, and IL-8 in both articular chondrocytes and synovial fibroblasts. PYC attenuated MSU crystal induced iNOS gene expression and NO production in chondrocytes. Activation of NF-κB and SAPK/JNK, ERK1/2 and p38 MAP kinases by MSU crystals in articular chondrocytes and synovial fibroblasts in vitro was attenuated by treatment with PYC. The acute inflammatory cell infiltration and increased expression of COX-2 and iNOS in synovial tissue and articular cartilage following intra-articular injection of MSU crystals in a rat model was inhibited by co-administration of PYC. PYC attenuated IL-1β induced COX-2 and MMP-9 expression in chondrocytes by real-time PCR, western blot and gelatin zymography. Activation of p38 MAP kinase by IL-1β in articular chondrocytes in vitro was attenuated by treatment with PYC. PYC also attenuated IL-1β and MSU crystals induced COX-2 in infrapatellar fat pad, as well as p38 MAP kinase. Conclusions This study demonstrates that PYC may be of value in treatment of MSU crystal-induced arthritis through its anti-inflammatory and anti-nitrosative activities. The articular chondrocytes treated by IL-1β is regarded as an in vitro osteoarthritic model. Our result may provide possible biologic mechanisms in satisfied results of PYC treatment to osteoarthritic patients. For infrapatellar fat pad, PYC has the similar therapeutic effects. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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