흰쥐 연골세포에서 campylaephora hypnaeoides 추출물의 연골보호 효과

Abstract

Osteoarthritis (OA) is the most common joint disease in people over the age of 65 and is characterized by gradual and irreversible cartilage wear, synovial inflammation, and subchondral bone remodeling. Since OA has no early symptoms and articular cartilage is a tissue with limited self-healing, it is very important to manage and maintain it healthy. Since OA has no early symptoms and is a tissue with limited self-healing, it is very important to manage and maintain it healthy. In order to maintain healthy joints, the intake of joint health functional foods is increasing, but glucosamine and MSM, which account for 80% of the raw materials of joint health functional foods, are raising concerns about efficacy. Therefore, this study aims to develop materials that are safe and have significant efficacy and provide scientific basis for joint health. Campylaephora hypnaeoides (Egonori, C. hypnaoides) is an edible seaweed belonging to the red algae, and is distributed along the coasts of Korea and Japan. C. hypnaoides is rich in polysaccharides, and has been consumed after being boiled since ancient times. Recently, it has been reported that C. hypnaoides ethanol extract reduced body fat and alleviates metabolic disorders through anti-inflammatory and anti-oxidant activity. However, no other functionalities have been reported. Therefore, in this study, the chondroprotective effect and its mechanism of C. hypnaoides fermented ethanol extract (FeCH) were analyzed using primary chondrocytes and osteoarthritis-induced experimental animals. C. hypnaoides was extracted using 30% fermented ethanol. The effect of FeCH on chondrocyte viability was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5- iphenyltetrazolium bromide (MTT) assay, which showed no cytotoxicity at 2 mg/ml. Pretreatment with FeCH in IL-1β-stimulated chondrocytes significantly inhibited the accumulation of nitric oxide (NO) and prostaglandin E2 (PGE2), which was analyzed using the ELISA assay. In addition, protein expression levels of inflammatory-related factors, such as inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2, interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and cartilage degrading-related enzymes, such as metalloproteinases (MMP)-1, -3, -13 and A disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-4, -5, were significantly decreased in IL-1β-stimulated chondrocytes pretreated with FeCH, were analyzed using western blot analysis. In addition, as a result of analyzing the content of type II collagen and proteoglycan through western blot analysis and alcian blue staining, FeCH pretreatment prevented the degradation of collagen type II and proteoglycan. It was analyzed through western blot analysis that the chondroprotective effect of FeCH may be mediated through MAPK and NF-κB signaling mechanisms. In an in vivo study, a rat model with damaged medial meniscus (DMM) was used as an osteoarthritis experimental animal model and orally administered daily for 8 weeks after surgery. At the end point, knee joints were collected and histologically analyzed with safranin O staining. As a result, articular cartilage was significantly protected in the FeCH group compared to the DMM group, without a change in body weight. These results suggested that C. hypnaeoides as a candidate material for the development of food and pharmaceutical materials for the treatment or prevention of degenerative arthritis.