Abstract
Osteoarthritis (OA) is the common form of arthritis and is characterized by disability and cartilage degradation. Although natural product extracts have been reported to have anti-osteoarthritic effects, the potential bioactivity of Ryupunghwan (RPH), a traditional Korean medicinal botanical formula that contains Astragalus membranaceus, Turnera diffusa, Achyranthes bidentata, Angelica gigas, Eclipta prostrata, Eucommia ulmoides, and Ilex paraguariensis, is not known well. Therefore, the inhibitory effects of single compounds isolated from RPH on the OA-related molecules were investigated using IL-1β-stimulated chondrosarcoma SW1353 (SW1353) cell model. Two bioactive compounds, isomucronulatol 7-O-β-d-glucoside (IMG) and ecliptasaponin A (ES) were isolated and purified from RPH using column chromatography, and then the structures were analyzed using ESI-MS, 1H-NMR, and 13C-NMR spectrum. The expression or amount of matrix metalloproteinase 13 (MMP13), COX1/2, TNF-α, IL-1β or p65 was determined by RT-PCR, Western blot, and enzyme-linked immunosorbent assay (ELISA). RPH pretreatment reduced the expression and amounts of MMP13, and the expression of collagen II, COX1/2, TNF-α, IL-1β or p65, which were increased in IL-1β-stimulated SW1353 cells. IMG reduced the expression of all OA-related molecules, but the observed inhibitory effect was less than that of RPH extract. The other single compound ES showed the reduced expression of all OA-related molecules, and the effect was stronger than that in IMG (approximately 100 fold). Combination pretreatment of both single components remarkably reduced the expression of MMP13, compared to each single component. These synergic effects may provide potential molecular modes of action for the anti-osteoarthritic effects of RPH observed in clinical and animal studies.
Highlights
The natural product formula popular in Asian countries, Ryupunghwan (RPH), is expected to provide pain relief and reduce inflammation as a health supplement
We examined whether RPH has an effect on the expression of COX1/2, which are known as a molecules (MMP13, TNF-α, IL-1β, IκBα, and COX1/2) related to cartilage degradation and regulator of inflammation [21]
NC, negative control; IL-1β, IL-1β-stimulated SW1353 cells; Gin, this experimental groups pretreated with RPH
Summary
The natural product formula popular in Asian countries, Ryupunghwan (RPH), is expected to provide pain relief and reduce inflammation as a health supplement. RPH contains Astragalus membranaceus, Turnera diffusa, Achyranthes bidentata, Angelica gigas, Eclipta prostrata, Eucommia ulmoides, and Ilex paraguariensis. Astragalus membranaceus contains calycosin and is a known Asian medicinal herb traditionally used for the treatment of several diseases, such as hypertension, cirrhosis or cancer therapy through anti-inflammatory and anti-carcinogenic properties, respectively [1]. Turnera diffusa, which contains a major constituent arbutin, possesses anti-ulcer activity, which could be attributed to the inhibition of lipid peroxidation, immunomodulatory and anti-oxidant activities [2]. Achyranthes bidentata has been shown to protect rat articular chondrocytes against interleukin-1β-induced inflammation and apoptosis in vitro [3]. Eucommia ulmoides ameliorates arthritis through inhibition of pro-inflammatory cytokines, and through reducing the degradation of cartilage and bone in rat collagen-induced arthritis [6]
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