基于尿液代谢组学技术的二妙散抗类风湿关节炎作用机制研究

Mechanism of Ermiao power in the treatment of rheumatoid arthritis based on urine metabolomics technology

目的采用高通量尿液代谢组学技术研究二妙散抗类风湿关节炎(rheumatoid arthritis, RA)的作用机制。方法将大鼠按随机数字表法分为空白组、模型组、给药组,每组8只。模型组与给药组皮内注射完全弗氏佐剂制备RA大鼠模型。给药组大鼠灌胃0.108 g/ml二妙散溶液,空白组与模型组灌胃等体积生理盐水,1次/d,连续给药14 d。分别于造模后第1、2、14天测定大鼠足跖肿胀度。采集大鼠尿液样本,并进行代谢组学分析。采用无监督和有监督的模式识别技术对获取到的高通量生物学信息数据进行全谱分析和降维处理,筛选和聚焦与RA相关的代谢信息,用以阐明RA的发病机制及二妙散抗炎的作用机制。结果造模后14 d,给药组大鼠足肿胀程度[(1.93±0.11)分比(2.36±0.19)分]较模型组减轻(P<0.01)。代谢轮廓结果显示,空白组大鼠与模型组大鼠代谢分布显著分离,且给药组处于二者之间。通过数据库匹配鉴定了13个与RA相关的生物标记物,分别为亚麻酸、花生四烯酸、5,6-EET、a-乳糖、蔗糖、海藻糖、前列腺素类、白三烯C4等。涉及亚油酸代谢、花生四烯酸代谢、淀粉和半乳糖代谢、鞘脂类代谢等代谢通路。结论二妙散可减轻RA大鼠炎症反应,通过调节亚油酸代谢、花生四烯酸代谢、淀粉和半乳糖代谢等发挥抗炎作用。

Objective To study the anti-rheumatoid arthritis mechanism of Ermiao powder by high-throughput urine metabolomics. Methods The rats were randomly divided into control group, model group and administration group with 8 rats in each group. The rat model of rheumatoid arthritis was established by intradermal injection of complete Freund’s adjuvant. Rats in control group were given Ermiao power solution 0.108 g/ml by gavage. The degree of joint swelling in rats was observed and scored. On this basis, metabolic data of rat urine samples were collected for metabolomic analysis. Unsupervised and supervised pattern recognition technology was used to analyze the high-throughput biological information data and reduce the dimension. Metabolic information related to rheumatoid arthritis was screened and focused to clarify the pathogenesis of rheumatoid arthritis and the therapeutic mechanism of Ermiao power. Results Compared with the model group, the swelling degree of the foot (1.93 ± 0.11 vs. 2.36 ± 0.19) in Ermiao power group significantly decreased (P<0.01). Metabolic profiles showed that the metabolic distribution of healthy rats was significantly separated from that of model rats, and the treatment group was in the middle of the two groups. From the macro-metabolic point of view, the metabolism of model rats changed dramatically. The Ermiao power had a good intervention effect on rheumatoid arthritis. Thirteen biomarkers related to rheumatoid arthritis were identified by database matching, including linolenic acid, arachidonic acid, 5,6-EET, alpha-lactose, sucrose, trehalose, prostaglandins and leukotriene C4. It involved linoleic acid metabolism, arachidonic acid metabolism, starch and galactose metabolism and sphingolipid metabolism. Conclusions The Ermiao power has significant therapeutic effect on rheumatoid arthritis rats. Regulation of the linoleic acid metabolism, arachidonic acid metabolism, starch and galactose metabolism may be the mechanism of its treatment of rheumatoid arthritis.