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KMID : 1123920190330020131
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Korean Journal of Oriental Physiology and Pathology
2019 Volume.33 No. 2 p.131 ~ p.140
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Concentration-dependent in vitro Anti-osteoarthritis Effects of Mixed Formula ? Pomegranate Concentrate Powder: Eucommiae Cortex: Achyranthis Radix 5:4:1 (g/g) on the Primary Cultured Rat Articular Chondrocytes
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Choi Beom-Rak
Ku Sae-Kwang
Kang Su-Jin
Park Hye-Rim
Sung Mi-Sun
Lee Young-Joon
Park Ki-Moon
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Abstract
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The objective of present study is to evaluate concentration-dependent in vitro anti-osteoarthritic (OA) effects of synergic mixed formula consisted of dried pomegranate juice concentrate powder, Eucommiae Cortex aqueous extract and Achyranthis Radix aqueous extract 5:4:1 (g/g) mixture on the primary cultured rat articular chondrocytes. First, any cytotoxic effect of mixture was observed using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium Bromide) assay. Next, cyto-protective effect of test substances was evaluated by using the recombinant human interleukin (rhIL)-1α induced chondrocytes. In addition, anti-inflammatory effects were also observed on the lipopolysaccaride (LPS) treated chondrocytes through prostaglandin E2 (PGE2) productions and 5-lipoxygenase (LPO) activities, and inhibitory effects on matrix metalloproteinase (MMP)-2 and MMP-9 activities were observed on rhIL-1α treated chondrocytes with their extracellular matrix (ECM) related mRNA expressions. No obvious cytotoxic effects of mixture were demonstrated. Inflammatory damages of chondrocytes and related ECM degradations induced by treatment of LPS or rhIL-1α were significantly and concentration-dependently inhibited by pretreatment of mixture from a concentration level of 0.001 mg/ml to 1 mg/ml. In addition, mixture showed IC50 for rhIL-1α-induced MMP-2 and MMP-9 activities as 44.01 and 162.47 μg/ml, and also showed EC50 for rhIL-1α-induced inhibition of collagen type II, SOX9 and aggrecan mRNA expression as 8.61, 10.79 and 4.47 μg/ml, respectively. It is observed that mixture showed concentration-dependent anti-inflammatory and cytoprotective ECM preserved effects on the primary cultured rat articular chondrocytes without cytotoxicity.
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KEYWORD
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Dried pomegranate juice concentrate powder, Eucommiae Cortex, Achyranthis Radix, Primary cultured rat articular chondrocytes, Osteoarthritis
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