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基于 UHPLC-Q-TOF/MS 联合多元统计分析的急性痛风性关节炎大鼠痛风发作期血浆代谢轮廓分析。

Plasma Metabolic Profiling Analysis of Gout Party on Acute Gout Arthritis Rats Based on UHPLC-Q-TOF/MS Combined with Multivariate Statistical Analysis.

机构信息

Tianjin University of Traditional Chinese Medicine, No. 10, Poyang Lake Road, West Zone, Tuanbo New City, Jinghai District, Tianjin 300193, China.

Tianjin Key Laboratory of Exercise Physiology and Sports Medicine, Institute of Sports and Health, Tianjin Sport University, No. 16, Donghai Road, Tianjin 300193, China.

出版信息

Int J Mol Sci. 2019 Nov 15;20(22):5753. doi: 10.3390/ijms20225753.

Abstract

Gout Party is a Chinese medicine prescription composed of , and , which can relieve joint pain caused by gouty arthritis (GA) and rheumatoid, and has a therapeutic effect on acute gouty arthritis (AGA). However, little information is available on the molecular biological basis and therapeutic mechanism of Gout Party for the treatment of AGA. AGA model was established by injecting sodium urate, and colchicine served as a positive control drug. We established a metabolomic method based on ultra-high-performance liquid chromatography-tandem quadrupole/time-of-flight mass spectrometry (UHPLC-Q-TOF/MS) to analyze the plasma samples of model group rats and blank group rats. Multiple statistical analyses, including principal component analysis (PCA) and partial least square discrimination analysis (PLS-DA), were used to examine metabolite profile changes in plasma samples. Finally, we identified 2-ketobutyric acid, 3-hexenedioic acid, but-2-enoic acid, and so on; 22 endogenous metabolites associated with AGA. After successful molding, we found that 2-ketobutyric acid, 3-hexenedioic acid, but-2-enoic acid, argininic acid, galactonic acid, lactic acid, equol 4'-O-glucuronide, deoxycholic acid glycine conjugate, glycocholic acid, sphinganine 1-phosphate, LPE (0:0/20:3), LPE (0:0/16:0), LPC (15:0) decreased significantly ( < 0.05 or < 0.01), alanine, erythrulose, 3-dehydrocarnitine, m-methylhippuric acid, 3-hydroxyoctanoic acid, p-cresol sulfate, estriol 3-sulfate 16-glucuronide, 10-hydroxy-9-(phosphonooxy)octadecenoate, docosahexaenoic acid increased significantly ( < 0.05 or < 0.01). After Gout Party treatment, 14 biomarkers had a tendency to normal conditions. These above biomarkers were mainly involved in fatty acid metabolism, bile acid metabolism, amino acid metabolism, and energy metabolism pathways. These results suggested that Gout Party exerted therapeutic effects of treating AGA by improving energy metabolism disorder and amino acid metabolism dysfunction, and attenuating fatty acid metabolism abnormal and inflammation. The results of this experiment provided a reference for revealing the metabolic mechanism produced by Gout Party in the treatment of AGA, but the subsequent studies need to be further improved and supported by relevant cell experiments and clinical experiments.

摘要

痛风方是由 、 、 等中药组成的方剂,能缓解痛风性关节炎(GA)和类风湿性关节炎引起的关节疼痛,对急性痛风性关节炎(AGA)有治疗作用。然而,对于痛风方治疗 AGA 的分子生物学基础和治疗机制,目前的信息还很少。我们通过尿酸钠注射建立 AGA 模型,秋水仙碱作为阳性对照药物。我们建立了一种基于超高效液相色谱-串联四极杆/飞行时间质谱(UHPLC-Q-TOF/MS)的代谢组学方法来分析模型组大鼠和空白组大鼠的血浆样本。采用主成分分析(PCA)和偏最小二乘判别分析(PLS-DA)等多种统计分析方法,对血浆样本的代谢谱变化进行了研究。最后,我们鉴定出 2-酮丁酸、3-己烯二酸、丁烯二酸等 22 种与 AGA 相关的内源性代谢物。成功造模后,我们发现 2-酮丁酸、3-己烯二酸、丁烯二酸、精氨酸、半乳糖醛酸、乳酸、雌马酚 4'-O-葡萄糖醛酸苷、脱氧胆酸甘氨酸缀合物、甘胆酸、鞘氨醇 1-磷酸、LPE(0:0/20:3)、LPE(0:0/16:0)、LPC(15:0)显著降低( < 0.05 或 < 0.01),丙氨酸、赤藓糖、3-脱氢肉碱、m-甲基马尿酸、3-羟基辛酸、对甲酚硫酸盐、雌三醇 3-硫酸盐 16-葡萄糖醛酸苷、10-羟基-9-(膦氧)十八烯酸、二十二碳六烯酸显著升高( < 0.05 或 < 0.01)。经痛风方治疗后,14 种生物标志物有向正常条件转变的趋势。以上生物标志物主要涉及脂肪酸代谢、胆汁酸代谢、氨基酸代谢和能量代谢途径。这些结果提示,痛风方通过改善能量代谢紊乱和氨基酸代谢功能障碍,减轻脂肪酸代谢异常和炎症,发挥治疗 AGA 的作用。本实验结果为揭示痛风方治疗 AGA 产生的代谢机制提供了参考,但后续研究还需要通过相关细胞实验和临床实验进一步完善和支持。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2a9/6888674/7c2a126b5842/ijms-20-05753-g001.jpg

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