Anti-inflammatory and anti-osteoarthritis effects of fermented Achyranthes japonica Nakai
Introduction
Osteoarthritis (OA) is the most disorder of joints and a leading cause of disability among the elderly population. The disease is characterized by progressive loss of articular cartilage and the formation of osteophytes, which lead to chronic pain and functional restrictions in the affected joints. Different factors can be involved in the development of OA, and while traumatic events are often causative, there are other factors such as genetic predisposition, defective positioning of joints, ageing, and malnutrition, which all lead to similar alterations in the joint cartilage (Swoboda, 2001).
Previous studies have shown that control of matrix metalloproteinases (MMPs) plays an important role in cartilage matrix turnover (Poole, 2000). Cartilage degradation is mediated by MMPs, and the increased release of MMP-3 may be associated with the development of OA (Lin et al., 2004, Lohmander et al., 1994). The enzymatic activities of MMPs are specifically controlled by tissue inhibitors of matrix metalloprotease (TIMPs) (Yoshihara, 2000a)). In healthy cartilage, there is a homeostasis between MMPs and TIMPs, which is disturbed in OA. Also, cartilage destruction in OA is recognized to be induced by proinflammatory cytokines such as nitic oxide (NO), prostaglandin E2 ( PGE2), and interleukin (IL)-1β, and tumor necrosis factor α (TNF-α) (Martel-Pelletier, 1998).
Current treatments for the management of OA are not curative and do not reverse the degenerative process of OA. Non-steroidal anti-inflammatory drugs (NSAIDs) have been used for the past several years in the treatment of OA, but their therapeutic effects remain unsatisfactory (Schnitaer et al., 2004). The prolonged consumption of synthetic NSAIDs is associated with serious adverse side effects, such as cardiovascular risks and gastrointestinal diseases (Bombardier et al., 2000, Farkouh et al., 2007, Lichtenberger et al., 2006). Accordingly, the development of novel drugs from natural sources, which can be safely used in prolonged treatment of OA and provide cartilage protection, is needed.
Achyranthes japonica Nakai (AJN) (Amaranthaceae) is used in traditional medicines or folk remedies in Korea, Japan, and China. In Korea, AJN has been widely used to control pain and improve dysfunction in OA patients (Han et al., 2005a). AJN contains several important phytochemicals such as saponins, inokosterone, ecdysterone, and oleanolic acid bisdesmoside (Han and Lee, 1991, Ida et al., 1994a). AJN extracts have been known to have anti-inflammatory, antioxidant, hepato-protective, and anti-cancer activities (Ida et al., 1994b, Meng and Li, 2001). However, few studies have been conducted on scientific approach to support for anti-inflammatory and anti-arthritis actions of AJN.
Several herbs have long been processed via microbial fermentation. Many studies have demonstrated that fermentation not only alters the original bioactivities of herbs, resulting in new treatment effects, but also enhances the original treatment efficacy (Miyake et al., 2005, Nakano et al., 2006, Lin and Chiang, 2008). In addition, fermentation by bacterial strains and lactobacillus have widely been used the production of high value-added phytochemicals from crude medicinal herbs (Eum et al., 2011). We have recently been developed the fermentation technology for improving biological activity of the crude medicinal drugs. In particular, fermentation of AJN by Bacillus sp. was found to increase the biological activity of AJN. This led to investigate anti-inflammatory and anti-osteoarthritis effects of fermented AJN.
The objectives of the present study were to investigate anti-inflammatory and anti-osteoarthritis activities of FAJN. First, anti-inflammatory activity between non-fermented AJN (NFAJN) and fermented AJN (FAJN) was compared. Anti-inflammatory activity of NFAJN and FAJN was evaluated by in vitro assay using LPS-induced Raw 264.7 cell. In addition, their cartilage protective effects were also determined in vitro assay using SW1353 cell and in vivo model system using collagenase-induced arthritis (CIA) in rabbits. Moreover, we isolated and identified 20-HES as a maker componenet in FAJN, and investigated quantitative changes of 20-HES in the AJN during fermentation using Bacillus subtilis.
Section snippets
Preparation of FAJN extract
Achyranthes japonica Nakai (AJN) was purchased from an oriental herb market in Seoul, Korea. A voucher sample was deposited in NUC company and authenticated by Professor Sang-Won Choi, Catholic University of Daegu. The procedure for preparing fermented AJN (FAJN) was as follows: AJN was mixed with 1.5 volume of water and autoclaved for 20 min at 121 °C. The solid-state fermentation of sterilized AJN was performed by inoculating with a 5% weight of Bacillus subtilis NUC1 (KCCM 10839P) and
Effects of FAJN on NO and PGE2 production in vitro
It is known that inflammatory mediators such as NO and PGE2 play key roles in the progression of cartilage destruction in OA. Therefore, we examined that the effects of NFAJN and FAJN on the level of NO and PGE2 in LPS-stimulated RAW 264.7 cells. Both NFAJN and FAJN considerably inhibited the production of NO and PGE2 in LPS-induced RAW 264.7 cells in a dose-dependent manner at a range of 10–50 μg/ml, and the inhibitory effects of FAJN were higher than those of NFAJN (Fig. 1). In addition,
Discussion
AJN, which is widely distributed throughout Far East countries, has been used in traditional folk medicine for a long time (Jeong, 1998). AJN has been reported to contain important phytochemicals including saponins (oleanolic acid glycoside etc.), caffeic acid, polysaccharides, insect molting hormones (inokosterone, ecdysterone, and ponasteroside A, etc.), K-succinate, K-oxalate, betain hydrate, and rubrosterone (Kim et al., 2006, Kim et al., 2008), which exert pharmacological activities such
Acknowledgment
This work was supported by the Regional Industry Technology Development project during the years 2008-2010 (Project No. 70004130).
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