Abstract
Objectives: This study investigated the anti-osteoarthritic effects of Kyejiinsam-tang (hereinafter referred to KIT) on the monosodium iodoacetate (MIA)-induced osteoarthritis rats. Methods: <in vitro> Anti-oxidative effects of KIT were measured by scavenging activities of DPPH, reactive oxygen species (ROS) and nitric oxide (NO). Scavenging activities of anti-oxidation in lipopolysaccharide (LPS)-treated RAW 264.7 cells were also measured for inhibitory effects against the production of inflammatory mediators (tumor necrosis factor-α, interleukin-1β, interleukin-6). <in vivo> Osteoarthritis was induced in rats by injecting MIA in the knee joint. Rats were divided into a total of 4 groups (n=6). The normal group were not treated at all without inducing osteoarthritis whereas the control group were induced for osteoarthritis by MIA and oral medicated physiological saline per day. The positive comparison group was injected with MIA and after 7 days, 2 mg/kg of Indomethacin. The experimental group was injected with MIA and after 7 days was medicated with 34 mg/kg of KIT. Indomethacin and KIT were orally-medicated for each substance a total of 4 weeks, once per day. Weight-bearing on hind legs was measured every week after MIA injection. At the end of the experiment (5 weeks after MIA injection), micro CT (computed tomography)-arthrography and histopathological examinations on the articular structures of knee joint were performed. The effect on inflammatory cytokines and immunological cells in synovial fluid was measured. Volume of cartilage was measured by micro CT-arthrography. Injury to synovial tissue was measured by H & E (hematoxylin and eosin), Safranin-O immunofluorescence. Results: Cytotoxicity against hFCs was insignificant. KIT showed the potent full term for DPPH. <in vitro> NO was significantly reduced by KIT (at 100, 200 μg/ml) and ROS was also reduced, but not significantly, by KIT (at 200 μg/ml). IL-6 and IL-1β were significantly reduced by KIT (at 100, 200 μg/ml) and TNF-α was also reduced, but not significantly, by KIT (at 200 μ/ml). <in vivo> In hind legs weight-bearing measurement, level of weight increased. Functions of liver and kidney were not affected. IL-1β was significantly reduced and TNF-α, IL-6 were also reduced but not significantly. PGE2 (prostaglandin E2), LTB4 (leukotriene B4) were significantly reduced in the KIT group. MMP-9 (matrix metalloproteinase-9), TIMP-1 (tissue inhibitor of metalloproteinases-1) and Osteocalcin were significantly reduced in the KIT group. Destruction of cartilage on micro CT arthrography was reduced but had no significant differences. Histopathologically, injury to synovial membrane of the KIT group was decreased and proteoglycan content of KIT group was increased. Conclusions: According to this study, Kyejiinsam-tang has inhibiting effect on the progression of arthritis in MIA-induced osteoarthritis rat. Kyejiinsam-tang has anti-oxidants and anti-inflammation effects, and is related to inhibiting the activity of inflammatory cytokine and injury of volume in cartilage. Keywords: Kyejiinsam-tang, osteoarthritis, anti-inflammation, anti-oxidation, MIA-induced
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