基于代谢组学技术的“温经活血”外治法早期干预膝骨关节炎机制研究

Knee osteoarthritis (KOA) is a common degenerative disease, contributing to higher disability rate. However, due to poor understanding of pathological mechanism, there was always a lack of effective method to treat KOA. Previously, our large sample, randomized controlled clinical trial confirmed that the external therapy of “warming channels activating blood circulation” has a good curative effect on KOA patients with different period. Following animal experiments also showed that the external therapy was capable of improving pain behavior in KOA rats and ameliorating synovitis and cartilage degeneration, in the in mid-late period of KOA. However, little studies had involved the early intervention effect of the external therapy of “warming channels activating blood circulation”. . In the pre-experiment, our study team used metabonomics to preliminarily evaluate the metabolic network of KOA patients in the early period. Finally, more than 20 different metabolites were identified. They were mainly referred to lipid metabolism, phospholipid and sheath metabolism. These results provided a basis for revealing the early intervention effect of the external therapy of “warming channels activating blood circulation”.. Consequently, this project intends to screen differential metabolite in KOA, validate the potential markers and related metabolic pathways, and clarify the therapeutic mechanism of the external therapy of “warming channels activating blood circulation” through animal and population study with metabonomics technique. The proposed research provides a new approach for the early prevention and treatment and a new evidence for curative effect of the external therapy of “warming channels activating blood circulation” on KOA.

膝骨关节炎（KOA）是骨科常见退行性疾病，致残率高，其病理机制尚不明确，缺乏有效治疗手段。前期大样本、随机对照临床研究证实了“温经活血”外治法对不同病程KOA患者具有良好疗效；后续动物实验亦表明“温经活血”外治法对KOA大鼠疼痛行为学、滑膜炎症及软骨退变等中后期病变具有较好的改善作用，但未涉及“温经活血”外治法的早期干预效应。.在预实验中，课题组采用代谢组学技术初步评估了早期KOA患者的代谢调控网络，共筛选出20余种差异代谢物，主要涉及脂质代谢、细胞磷脂以及鞘质代谢等代谢通路，为揭示“温经活血”外治法的早期干预效应提供了基础。.为此，本课题进一步拟在动物模型及人群研究中采用代谢组学技术筛选KOA差异代谢物，验证其潜在标志物，阐述相关代谢通路，并结合分子生物学实验从代谢网络调控的角度阐明“温经活血”外治法早期干预KOA的疗效机制，从而实现病程全覆盖。

KOA是骨科常见退行性疾病，致残率高，其病理机制尚不明确，缺乏有效治疗手段。代谢组学的整体性、动态性与中医的整体观、动态诊疗观念十分契合，代谢组学通过对机体内源性代谢物的分析，寻找差异性代谢物，从整体上反映机体的代谢情况，对疾病的诊断及药物的作用靶点具有十分重要的指导意义。.本研究先通过动物和临床研究观察温经活血法的临床疗效，再通过代谢组学技术筛选KOA差异代谢物及温经活血外治法作用靶点，阐述相关代谢通路，结合分子生物学实验从代谢网络调控的角度阐明温经活血外治法干预各阶段KOA的疗效机制。.研究结果显示温经活血外治法能显著改善KOA滑膜炎症，延缓软骨退变，还能显著提高KOA大鼠冷痛阈值和机械痛阈值。动物样本GC-MS结果显示温经活血外治法能较好回调KOA大鼠血清中的代谢紊乱，各阶段相关代谢通路有所不同，主要与精氨酸和脯氨酸代谢，精氨酸生物合成，嘌呤代谢，D-谷氨酰胺和D-谷氨酸代谢，苯丙氨酸、硌氨酸和色氨酸的生物合成，丙氨酸、天冬氨酸和谷氨酸，硌氨酸代谢关系密切。LC-MS显示温经活血外治法能回调各阶段KOA大鼠血清中的差异代谢物，主要与磷脂酰胆碱、溶血磷脂酰胆碱、心磷脂、神经酰胺等关系密切。.临床研究结果显示温经活血外治法能显著降低KOA患者WOMAC评分，改善关节功能分级。临床样本GC-MS结果显示温经活血法能较好回调KOA患者各阶段血清中的代谢紊乱，主要与甘氨酸、丝氨酸和苏氨酸代谢，氨酰基-tRNA生物合成，半胱氨酸和蛋氨酸代谢，丙氨酸、天冬氨酸和谷氨酸，精色氨酸代谢等关系密切。LC-MS结果显示温经活血外治法对KOA各阶段回调代谢产物有所不同，主要与二酰基甘油、三酰基甘油、磷脂酰胆碱、溶血磷脂酰胆碱、心磷脂、神经酰胺等关系密切。.本课题拟通过动物和临床试验观察温经活血外治法对KOA的干预效应，并通过代谢组学技术筛选相关生物标志物，从代谢角度初步探讨温经活血外治法治疗各阶段KOA的作用机制，为中医外治法对KOA的治疗提供新思路。