SKI306X inhibition of glycosaminoglycan degradation in human cartilage involves down-regulation of cytokine-induced catabolic genes

BACKGROUND/AIMS: SKI306X, a mixed extract of three herbs, Clematis mandshurica (CM), Prunella vulgaris (PV), and Trichosanthes kirilowii (TK), is chondroprotective in animal models of osteoarthritis (OA). The objectives of this study were to investigate its effect on interleukin (IL)-1beta-induced degradation of glycosaminoglycan (GAG) and the basis of its action in human OA cartilage, as well as to screen for the presence of inhibitors of matrix metalloproteinase (MMP)-13 and a disintegrin and metalloprotease with thrombospondin motifs (ADAMTS)-4 in SKI306X and its component herbs, as well as in fractions from SKI306X. METHODS: Human OA chondrocytes and cartilage explants were obtained during total knee replacements and incubated with IL-1beta +/- oncostatin M with or without SKI306X or its component herb extracts. GAG degradation was assayed in cartilage explants using a commercial kit. Expression of genes involved in cartilage destruction was measured by real-time polymerase chain reaction using chondrocyte RNA. SKI306X was fractionated by preparative liquid chromatography to test for the presence of inhibitors of MMP-13 and ADAMTS-4. RESULTS: SKI306X and PV inhibited IL-1beta-induced GAG release from cartilage explants, and SKI306X, CM, PV, and TK inhibited IL-1beta-induced MMP gene expression. Unexpectedly, SKI306X greatly stimulated IL-1beta + oncostatin M-induced ADAMTS-4 gene expression, probably due to its TK component. Some fractions of SKI306X also inhibited ADAMTS-4 activity. CONCLUSIONS: SKI306X and its herbal components inhibit GAG degradation and catabolic gene expression in human OA chondrocytes and cartilage explants. SKI306X likely also contains one or more ADAMTS-4 inhibitor.

Dietary Flavonoid Apigenin is not Effective in Preventing Development of a Bleomycin-Induced Murine Model of Scleroderma

OBJECTIVE: Systemic sclerosis is a connective tissue disease characterized by vasculopathy, excessive accumulation of extracellular matrix, and fibrosis of the skin and internal organs. The dietary flavonoid apigenin has been shown to reduce expression of the myofibroblast phenotype and to inhibit contraction of collagen gels. We investigated the effect of apigenin on the prevention and treatment of a modified bleomycin-induced animal model of scleroderma. METHODS: Recently, we successfully induced scleroderma by weekly subcutaneous injections of bleomycin using a thermo-reversible combination gel composed of low molecular weight methylcellulose. A weekly subcutaneous injection of methylcellulose gel loaded with bleomycin induced focal skin fibrosis on the back skin and fibrotic phenotype of lung tissue in mice. The histologic examination of skin and lungs, collagen assay of lungs, and expression of connective tissue growth factor were investigated. RESULTS: Daily intra-peritoneal injection of 1.0 mg/kg or 2.5 mg/kg of apigenin starting a week before the bleomycin injections failed to prevent the development of skin fibrosis and reduce the fibrotic phenotypes of skin and lung tissue. CONCLUSION: Although some in vitro experiments have supported a potential role of apigenin in the treatment of fibrosis, dietary flavonoid apigenin is not effective in preventing development of a bleomycin-induced murine model of scleroderma.

Inhibition of the IL-1beta-induced Expression of Matrix Metalloproteinases by Controlled Release of IL-1 Receptor Antagonist Using Injectable and Thermo-reversible Gels in Human Osteoarthritis Chondrocytes

OBJECTIVE: IL-1beta is involved in the degradation of articular cartilage in various arthritides, including osteoarthritis (OA). Competitive inhibition of IL-1beta by IL-1 receptor antagonists (IL-1Ra) may represent a pathogenesis-based strategy for inhibiting degradation of the cartilage matrix. We investigated the hypothesis that controlled release of IL-1Ra using injectable, thermoreversible and complex coacervate combination gels as drug delivery systems might reduce matrix degradation in OA. METHODS: Thermoreversible combination gels that can be injected into joints were formed in aqueous solution by making a complex coacervate with recombinant human IL-1Ra (anakinra) and cationic macromolecules, and this was followed by co-formulation with methylcellulose as a negative thermosensitive polysaccharide. Gels containing anakinra were positioned in the upper insert of a transwell system and human OA chondrocytes were placed in the lower compartment and then they were stimulated with IL-1beta. The expression of matrix metalloproteinases (MMPs) was examined by performing real time PCR and ELISA. RESULTS: Complex coacervation between anakinra and protamine was successfully completed. IL-1Ra was released from the gels in a sustained release pattern for extended periods with minimal initial bursts. IL-1beta markedly enhanced the expression of MMP. The IL-1Ra released from the gels significantly inhibited the IL-1beta-induced MMP expression in the chondrocytes. CONCLUSION: We developed and optimized a novel injectable and thermoreversible gel system for the controlled release of IL-1Ra, and this drug delivery system effectively inhibited the IL-1beta-induced MMP expression of chondrocytes in a transwell system. Intra-articular local delivery of injectable and thermoreversible gels containing IL-1Ra into knees has the potential to provide prolonged therapy based on the pathophysiology of knee OA.

Measurement of Purine Contents in Korean Alcoholic Beverages / 대한류마티스학회지

OBJECTIVE: Gout is one of the most common forms of inflammatory arthritides among men, which is caused primarily by chronic hyperuricemia. Although pharmacological therapy is the mainstay treatment to manage gout, limiting the consumption of dietary purine is also important. Several epidemiological studies have reported that alcohol consumption is closely related to hyperuricemia and gout. The objective of this study was to determine the purine content in common Korean alcoholic beverages using high performance liquid chromatography (HPLC) to provide a dietary guideline for those with hyperuricemia or gout. METHODS: Thirty-five alcoholic beverages were analyzed. Blindly labeled samples of each alcoholic beverage were degassed and frozen. The sample preparation prior to HPLC followed the methods of Japanese researchers. HPLC was performed to analyze adenine, guanine, hypoxanthine, and xanthine content in the alcoholic beverages. RESULTS: The standard curves were linear for all purines. Purine contents were as follows: beer (42.26~146.39 micromol/L, n=12), medicinal wine (8.2 and 40.41 micromol/L, n=2), rice wine (13.19 micromol/L), Makgeolri (11.71 and 24.72 micromol/L, n=2), red wine (0, 6.03, and 17.9 micromol/L, n=3). No purines were found in fruit wine (n=2), Kaoliang (n=1), white wine (n=1), or distilled alcoholic beverages, such as soju (n=10) or whiskey (n=1). CONCLUSION: Among popular Korean alcoholic beverages, beer contained a considerable amount of purines, whereas distilled alcoholic beverages did not. Patients with either gout or hyperuricemia should avoid alcoholic beverages, especially those containing large amounts of purines.

Induction of Animal Model of Scleroderma with Repeated Injection of Bleomycin / 대한류마티스학회지

OBJECTIVE: To induce a mouse model of scleroderma with repeated bleomycin injections for research into human scleroderma at our research laboratory. METHODS: The protocol of Yamamoto et al. was replicated to establish the bleomycin-induced mouse model of scleroderma. RESULTS: A mouse model of scleroderma was induced by repeated subcutaneous injections of bleomycin. The dermal thickness increased with homogeneous and thickened collagen bundles. Semiquantitative measurements of dermal fibrosis were prominent in bleomycin-treated mice. CONCLUSION: A mouse model of scleroderma was induced with repeated injections of bleomycin at our laboratory.