Pradit Waranee, Chomdej Siriwadee, Nganvongpanit Korakot, Ongchai Siriwan
Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai, 50200, Thailand.
In Vitro Cell Dev Biol Anim. 2015 Apr;51(4):336-44. doi: 10.1007/s11626-014-9846-y. Epub 2014 Dec 17.
Phyllanthus amarus Schum. & Thonn. (P. amarus) has been reported to exhibit anti-inflammation and antiarthritis properties leading to our interest to examine its beneficial effect in osteoarthritis. Thus, this study aimed to explore the chondroprotective potential of P. amarus extract (PAE) and its major compounds, phyllanthin and hypophyllanthin, in a cartilage explant model. Various concentrations of P. amarus extract, phyllanthin and hypophyllanthin, were treated on porcine articular cartilage explants induced with 25 ng/ml of interleukin-1 beta (IL-1β). After 4 days of incubation, the culture medium was measured for the release of sulfate glycosaminoglycans (s-GAGs) and matrix metalloproteinase-2 (MMP-2) activity by DMMB binding assay and zymography, respectively. The explant tissues were analyzed for the remaining of uronic acid content by colorimetric assay and stained with safranin-O for investigation of proteoglycan content. Cell viability of this model was evaluated by lactate dehydrogenase (LDH) assay. Chondroprotective potential of PAE and the major components against IL-1β-induced cartilage explant degradation were revealed by the decreased s-GAGs level and MMP-2 activity in culture medium consistent with an increase in uronic acid and proteoglycan contents in the explants when compared to the IL-1β treatment. These results agreed with those of diacerein and sesamin which used as positive controls. In addition, better chondroprotective activities of P. amarus crude extracts than those of the purified components were disclosed in this study. Hence, this is a pioneering study presenting the chondroprotective potential of PAE which may augment its application for therapeutic use as an antiarthritic agent.
苦味叶下珠（Phyllanthus amarus Schum. & Thonn.）已被报道具有抗炎和抗关节炎特性，这引发了我们研究其对骨关节炎有益作用的兴趣。因此，本研究旨在探讨苦味叶下珠提取物（PAE）及其主要化合物叶下珠素和去甲叶下珠素在软骨外植体模型中的软骨保护潜力。将不同浓度的苦味叶下珠提取物、叶下珠素和去甲叶下珠素处理用25 ng/ml白细胞介素-1β（IL-1β）诱导的猪关节软骨外植体。孵育4天后，分别通过DMMB结合测定法和酶谱法测量培养基中硫酸糖胺聚糖（s-GAGs）的释放和基质金属蛋白酶-2（MMP-2）的活性。通过比色法分析外植体组织中剩余的糖醛酸含量，并用番红O染色以研究蛋白聚糖含量。通过乳酸脱氢酶（LDH）测定法评估该模型的细胞活力。与IL-1β处理相比，培养基中s-GAGs水平和MMP-2活性降低，同时外植体中糖醛酸和蛋白聚糖含量增加，这表明PAE及其主要成分对IL-1β诱导的软骨外植体降解具有软骨保护潜力。这些结果与用作阳性对照的双醋瑞因和芝麻素的结果一致。此外，本研究还揭示了苦味叶下珠粗提物比纯化成分具有更好的软骨保护活性。因此，这是一项开创性研究，展示了PAE的软骨保护潜力，这可能会扩大其作为抗关节炎药物的治疗应用。
