目的利用 1H 核磁共振(nuclear magnetic resonance spectroscopy,NMR)技术对膝骨关节炎模型兔的血清及关节液进行检测,找出注射玻璃酸钠前后的代谢差异和联系,为临床骨关节炎的治疗提供理论和实验依据。方法选取 30 只新西兰大白兔,随机分为健康空白对照组、磷酸盐缓冲液注射组、玻璃酸钠注射组进行处理,每组各 10 只,术后 10 周行大体观察、影像学检查、组织学形态检查及对血清及关节液分别进行代谢组学 1H-NMR 检测及多维统计分析。结果大体观察、影像学检查、组织学形态检查均示玻璃酸钠注射组优于磷酸盐缓冲液注射组。代谢组学研究显示,玻璃酸钠注射组血清及关节液中代谢产物较磷酸盐缓冲液注射组、健康空白对照组均有变化,差异有统计学意义(P<0.05)。根据差异代谢物检索相关途径,分析发现玻璃酸钠治疗骨关节炎的作用可能与蛋白质的生物合成、氨基酸循环、丙酮酸代谢、糖异生等代谢通路有关。结论基于 1H-NMR 的代谢组学研究发现玻璃酸钠对骨关节炎的作用主要与蛋白质代谢、脂代谢和能量代谢途径的激活有关。该研究为玻璃酸钠治疗膝骨关节炎的具体机制研究提供了新思路和新依据。

ObjectiveTo detect the metabolites of the serum and joint fluid from rabbits’ osteoarthritis model with 1H nuclear magnetic resonance spectroscopy (NMR) technique, study the metabolic differences and connections of serum, synovial and cartilage of rabbits after the articular cavity injection of sodium hyaluronate, and explore osteoarthritis and metabolic mechanism in the process of treating sodium hyaluronate using sodium hyaluronate, thus provide new ideas and basis of the specific mechanisms in the treatment of osteoarthritis via sodium hyaluronate.MethodsWe selected 30 healthy New Zealand white rabbits, 6 months old, and randomly divided them into three groups as follows: blank control group, model phosphate buffer saline (PBS) liquid injection group and model injection of sodium hyaluronate group, with 10 rabbits in each group. Ten weeks after surgery, all experimental animals were put to death and observed in correlation studies regarding general condition, imaging examination, and histological examination. Metabolites 1H NMR detection and data preprocessing were performed in the serum and joint fluid samples.ResultsThe results considering general condition, general sample observation, imaging examination and histology indicated advantages in sodium hyaluronate group over PBS group. Metabolomics analysis showed statistically significant changes of metabolites in the serum and joint fluid compared with the PBS group and the blank control group (P<0.05). According to the relevant ways of differences metabolites retrieval, analysis found that the effect of sodium hyaluronate on osteoarthritis might be related to protein biosynthesis, amino acid circulation, the metabolic process of pyruvic acid, gluconeogenesis and other metabolic pathways.ConclusionsBased on the research of 1H-NMR metabolomics, the results suggest that the effect of sodium hyaluronate on osteoarthritis is mainly related with the activation of protein metabolism, abnormal lipid and energy metabolic pathways. This study provides new ideas and basis on the concrete mechanism in the treatment of knee osteoarthritis using sodium hyaluronate.

关键词: 骨关节炎; 玻璃酸钠; 核磁共振; 代谢组学; 多变量数据分析

Key words: Osteoarthritis; Sodium hyaluronate; Nuclear magnetic resonance; Metabolomics; Multivariate data analysis

引用本文: 赖思可, 曹畅, 李箭, 李棋. 玻璃酸钠作用于兔膝骨关节炎的代谢组学研究. 华西医学, 2018, 33(9): 1153-1161. doi: 10.7507/1002-0179.201808068 复制

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