Department of Medicine/Rheumatology, MetroHealth Medical Center, Case Western Reserve University, Cleveland, Ohio 44109, USA.
BMC Complement Altern Med. 2011 Aug 19;11:66. doi: 10.1186/1472-6882-11-66.
Conventional treatments for the articular diseases are often effective for symptom relief, but can also cause significant side effects and do not slow the progression of the disease. Several natural substances have been shown to be effective at relieving the symptoms of osteoarthritis (OA), and preliminary evidence suggests that some of these compounds may exert a favorable influence on the course of the disease. The objective of this study was to investigate the anti-inflammatory/chondroprotective potential of a Herbal and amino acid mixture containing extract of the Uncaria tomentosa, Boswellia spp., Lepidium meyenii and L-Leucine on the IL-1β-induced production of nitric oxide (NO), glycosaminoglycan (GAG), matrix metalloproteinases (MMPs), aggrecan (ACAN) and type II collagen (COL2A1) in human OA chondrocytes and OA cartilage explants.
Primary OA chondrocytes or OA cartilage explants were pretreated with Herbal-Leucine mixture (HLM, 1-10 μg/ml) and then stimulated with IL-1β (5 ng/ml). Effect of HLM on IL-1β-induced gene expression of iNOS, MMP-9, MMP-13, ACAN and COL2A1 was verified by real time-PCR. Estimation of NO and GAG release in culture supernatant was done using commercially available kits.
HLM tested in these in vitro studies was found to be an effective anti-inflammatory agent, as evidenced by strong inhibition of iNOS, MMP-9 and MMP-13 expression and NO production in IL-1β-stimulated OA chondrocytes (p < 0.05). Supporting these gene expression results, IL-1β-induced cartilage matrix breakdown, as evidenced by GAG release from cartilage explants, was also significantly blocked (p < 0.05). Moreover, in the presence of herbal-Leucine mixture (HLM) up-regulation of ACAN and COL2A1 expression in IL-1β-stimulated OA chondrocytes was also noted (p < 0.05). The inhibitory effects of HLM were mediated by inhibiting the activation of nuclear factor (NF)-kB in human OA chondrocytes in presence of IL-1β.
Our data suggests that HLM could be chondroprotective and anti-inflammatory agent in arthritis, switching chondrocyte gene expression from catabolic direction towards anabolic and regenerative, and consequently this approach may be potentially useful as a new adjunct therapeutic/preventive agent for OA or injury recovery.
传统的关节疾病治疗方法通常可以有效缓解症状，但也会引起明显的副作用，并且不能减缓疾病的进展。已经证明，几种天然物质对缓解骨关节炎（OA）的症状有效，并且初步证据表明，其中一些化合物可能对疾病的进程产生有利影响。本研究的目的是研究含有钩藤、乳香、玛卡和 L-亮氨酸提取物的草药和氨基酸混合物对白细胞介素-1β（IL-1β）诱导的一氧化氮（NO）、糖胺聚糖（GAG）、基质金属蛋白酶（MMPs）、聚集蛋白聚糖（ACAN）和 II 型胶原（COL2A1）产生的抗炎/软骨保护潜力在人 OA 软骨细胞和 OA 软骨外植体中。
用 Herbal-Leucine 混合物（HLM，1-10μg/ml）预处理原代 OA 软骨细胞或 OA 软骨外植体，然后用 IL-1β（5ng/ml）刺激。通过实时 PCR 验证 HLM 对 IL-1β诱导的 iNOS、MMP-9、MMP-13、ACAN 和 COL2A1 基因表达的影响。使用市售试剂盒测定培养上清液中 NO 和 GAG 的释放。
在这些体外研究中，HLM 被证明是一种有效的抗炎剂，因为它强烈抑制了白细胞介素-1β刺激的 OA 软骨细胞中的 iNOS、MMP-9 和 MMP-13 表达和 NO 产生（p<0.05）。支持这些基因表达结果，IL-1β 诱导的软骨基质降解，如软骨外植体中 GAG 的释放也显著阻断（p<0.05）。此外，在存在草药-亮氨酸混合物（HLM）的情况下，还观察到白细胞介素-1β刺激的 OA 软骨细胞中 ACAN 和 COL2A1 表达的上调（p<0.05）。HLM 的抑制作用是通过抑制核因子（NF）-kB 在人 OA 软骨细胞中的活化来介导的，在白细胞介素-1β存在的情况下。
我们的数据表明，HLM 可能是关节炎的软骨保护和抗炎剂，将软骨细胞的基因表达从分解代谢方向转向合成代谢和再生，因此，这种方法可能作为 OA 或损伤恢复的新辅助治疗/预防剂具有潜在的用途。
