基于UPLC-Q-TOF-MS代谢组学技术研究淫羊藿-川芎对骨关节炎大鼠的调控作用

目的 基于超高效液相色谱-四极杆-飞行时间质谱联用仪（ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry，UPLC-Q-TOF-MS）代谢组学技术探究骨关节炎大鼠的血浆代谢特征及淫羊藿-川芎配伍对骨关节炎大鼠代谢的调控作用。方法 SD大鼠随机分成正常组、假手术组、模型组、病理经时评价组、淫羊藿组、川芎组、淫羊藿-川芎组（40 g/kg）、阳性药物组（盐酸氨基葡萄糖0.12 g/kg＋塞来昔布1.00 g/kg）。模型组、病理经时评价组和各给药组采用交叉韧带-半月板损伤术建立骨关节炎模型，造模第6周各给药组给予相应药物干预，正常组、模型组给予等体积蒸馏水。采集0、2、4、6、8、10、12、14周大鼠血浆，通过病理组织学及血浆中基质金属蛋白酶-13（matrix metalloproteinase-13，MMP-13）检测结果评价药效，UPLC-Q-TOF-MS技术分析大鼠血浆代谢轮廓，并结合多元统计分析，检测各组大鼠血浆代谢相关生物标志物的变化。结果 造模4周后MMP-13水平明显升高（P<0.01），给药8周后显著下降（P<0.01）。基于代谢组学发现给药组大鼠血浆中2种潜在生物标志物发生明显变化，分别为溶血磷脂酰胆碱（16∶0）、磷脂酰胆碱（18∶0/20∶4），主要涉及磷脂代谢通路。结论 淫羊藿、川芎单用和两者配伍均可回调差异代谢物治疗骨关节炎大鼠，且配伍效果优于单用。

Effects of Epimedii Folium-Chuanxiong Rhizoma on osteoarthritic rats based on UPLC-Q-TOF-MS metabonomics

Objective To investigate the plasma metabolic profile of rats with osteoarthritis (OA) and regulatory effects of Yinyanghuo (Epimedii Folium)-Chuanxiong (Chuanxiong Rhizoma) on metabolism of osteoarthritic rats based on ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) metabonomics technology. Methods SD rats were randomly divided into normal group, sham group, model group, pathological evaluation group during the process of OA modeling, Epimedii Folium group, Chuanxiong Rhizoma group, Epimedii Folium-Chuanxiong Rhizoma group (40 g/kg), and positive drug group (glucosamine hydrochloride 0.12 g/kg + celebrex 1.00 g/kg). Osteoarthritis model was established by cruciate ligament-meniscus injury surgery in model group, pathological time evaluation group and administration groups, and corresponding drug interventions were given to each drug administration group at 6th week of modelling. Equal amount of distilled water was given to the normal and model groups. Plasma was collected from rats at 0, 2, 4, 6, 8, 10, 12 and 14 weeks. The drug efficacy was evaluated by pathological histology and the results of matrix metalloproteinase-13 (MMP-13) assay. The plasma metabolic profiles of rats were analyzed by UPLC-Q-TOF-MS technology, and changes in plasma metabolism-related biomarkers were detected by combining with multivariate statistical analyses in each group. Results The level of MMP-13 was increased after four weeks of modelling (P < 0.01) and decreased significantly after eight weeks of drug administration (P < 0.01). Based on metabonomics, two potential biomarkers were found to be significantly changed in the plasma of rats in administration group, namely lysophosphatidylcholine (16:0) and phosphatidylcholine (18:0/20:4), which were mainly involved in phospholipid metabolic pathway. Conclusion Epimedii Folium and Chuanxiong Rhizoma alone and the combination can reverse the differential metabolites for the treatment of osteoarthritis in rats, and the combination is more effective than the single use.