CHONDROPROTECTIVE ACTIVITY OF POMEGRANATE EXTRACT

描述（由申请人提供）：骨关节炎（OA）是美国和全世界老年人群中最常见的肌肉骨骼疾病。OA的特征在于手、膝、脊柱和髋关节的关节软骨退化。虽然已经确定了诱发OA发展的条件，但实际原因仍然未知，目前的治疗选择仅限于使用NSAIDS缓解症状。然而，使用非甾体抗炎药可能会因显着的副作用而变得复杂，包括胃肠道出血、心血管不良事件和肾毒性。石榴果（Punica granatum L）因其药用特性而备受推崇。印度的乌纳尼（Unani）和阿育吠陀（Ayurvedic）医学系统中广泛使用Puppelfur（PF）或其提取物（PFE）治疗糖尿病和结肠炎。PF的可食用部分是 富含花青素，一组多酚化合物，具有抗氧化和抗炎特性。我们先前报道PFE对IL-1诱导的MAPK亚群p38-MAPK和JNK、转录因子NF-κ B、B的表达以及体外培养的人软骨组织和软骨细胞中MMPs的表达。最近，我们发现（1）口服PFE可抑制小鼠胶原诱导性关节炎（CIA）的发生;（2）生物可利用的PFE成分/代谢物可抑制软骨细胞中考克斯-2活性和IL-1诱导的NO和PGE 2的产生。拟议的研究利用了这些新的发现，因为在这里，我们将扩展这些研究，并将确定体外研究结果与OA动物模型中体内PFE使用的相关性。我们将检验“口服PFE抑制软骨降解并抑制前交叉韧带横断（ACLT）诱导的兔膝关节OA进展”的假设。在目的-1中，我们将确定几种已知的PFE衍生的花青素和鞣花单宁（ET）的身份和浓度，这些花青素和鞣花单宁（ET）在给予不同剂量PFE的兔的血浆和滑液（SF）中变得生物可利用;在目的-2中，我们将确定和比较含有PFE衍生的ET和花青素的血浆和SF在抑制IL-1？- 体外诱导人和兔软骨外植体的分解代谢反应;在aim-3中，我们将使用上述研究中确定的最相关剂量喂养兔子，并评估口服PFE对ACLT模型中关节软骨变性的影响。OA。 公共卫生相关性：骨关节炎（OA）的特征是高水平的IL-1、ROS的过度产生和受影响关节的关节软骨退化。然而，其病因和确切的发病机制仍不清楚。在这里，我们将确定是否消耗石榴果（PFE）的标准化提取物可以减缓或延缓体内关节软骨退化。拟议研究的主要目的是（a）确定与PFE的软骨/软骨保护活性相关的体外分子机制;以及（B）使用广泛使用的人类OA动物模型测试体外研究与体内情况的相关性。我们的研究结果可能为开发治疗OA的新治疗策略打开大门。

DESCRIPTION (provided by applicant): Osteoarthritis (OA) is the most common musculoskeletal disease among the aging population in the United States and throughout the world. OA is characterized by degeneration of articular cartilage of the joints in hands, knees, spine and hips. While conditions predisposing to the development of OA have been identified, the actual causes remain unknown and the current treatment options are limited to relief of symptoms using NSAIDS. However, use of NSAIDS may be complicated by significant side effects that include gastrointestinal bleeding, cardiovascular adverse events and nephrotoxicity. Pomegranate fruit (Punica granatum L) is revered through the ages for its medicinal properties. Pomegranate fruit (PF) or its extract (PFE) is widely used in Unani and Ayurvedic medicinal systems in India for the treatment of diabetes and Colitis. Edible part of PF is rich in anthocyanins, a group of polyphenolic compounds that possess antioxidant and anti-inflammatory properties. We previously reported that PFE exert a potent inhibitory effect on IL-1¿-induced activation of MAPK sub-groups p38-MAPK and JNK, transcription factor NF-?B and the expression of MMPs by human cartilage explants and chondrocytes in vitro. Recently we have shown that (1) oral consumption of PFE inhibited the development of collagen-induced arthritis in mice(CIA); and (2) bioavailable PFE constituents/metabolites inhibited COX-2 activity and IL-1¿-induced production of NO and PGE2 in chondrocytes. Proposed studies capitalizes on these novel findings as here we will extend these studies and will determine the relevance of the in vitro findings to in vivo PFE use in an animal model of OA. We will test the hypothesis that "oral consumption of PFE inhibits cartilage degradation and suppresses the progression of knee OA in rabbits induced by anterior cruciate ligament transection (ACLT)". In aim-1 we will establish the identity and concentration of several of the known PFE-derived anthocyanins and ellagitanins (ET) that become bioavailable in plasma and synovial fluid (SF) in rabbits given different doses of PFE; in aim-2 we will determine and compare the efficacy of plasma and SF containing PFE-derived ET and anthocyanins in inhibiting the IL-1¿-induced catabolic responses in human and rabbit cartilage explants in vitro; in aim-3 we will use the dose determined to be most relevant from above studies for feeding rabbits and evaluate the effect of oral consumption of PFE on articular cartilage degeneration in the ACLT model of OA. Public Health Relevance: Osteoarthritis (OA) is characterized by high high levels of IL-1¿, excessive production of ROS and articular cartilage degeneration in the affected joints. However, its etiology and precise pathogenetic mechanisms remain unclear. Here, we will determine whether consumption of a standardized extract of pomegranate fruit (PFE) can slow or retard the artciular cartilage degeneration in vivo. The broad objectives of the proposed studies are (a) to identify the molecular mechanism in vitro associated with cartilage/chondroprotective activity of PFE; and (b) to test the relevance of in vitro studies to the in vivo situation using a widely used animal model of human OA. Our findings are likely to open the door for the development of novel therapeutic strategies for the treatment of OA.