摘要
该研究基于超高效液相色谱-串联四极杆飞行时间质谱(UHPLC-Q-TOF-MS)联用技术的代谢组学研究分析川牛膝对膝骨关节炎大鼠血清、尿液的影响,探究川牛膝治疗膝骨关节炎的作用机制。采用2%木瓜蛋白酶和0.03 mol·L-1半胱氨酸混合液构建膝骨关节炎模型,通过检测模型组血清中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和基质金属蛋白酶-3(matrix metalloproteinase-3,MMP-3)的含量初步评价川牛膝对膝骨关节炎大鼠的治疗作用。利用UHPLC-Q-TOF-MS技术建立检测正常组、模型组、阳性药组、川牛膝组大鼠样本中内源性小分子代谢物代谢轮廓谱,以偏最小二乘判别分析(PLS-DA)及正交偏最小二乘判别分析(OPLS-DA)等多元统计分析进行模式识别,结合t检验、变量投影重要性(VIP)和差异倍数(fold-change)筛选潜在生物标志物,并借助MetaboAnalyst 5.0数据库富集代谢通路。结果表明川牛膝可改善膝骨关节炎大鼠一般体征指标,并降低大鼠血清中TNF-α和MMP-3的含量。代谢组学结果显示最终从血清和尿液中共筛选出28个与川牛膝调控作用相关的潜在生物标志物,其主要涉及精氨酸生物合成,三羧酸循环,核黄素代谢,丙氨酸、天冬氨酸和谷氨酸代谢,乙醛酸和二羧酸代谢,嘧啶代谢,烟酸和烟酰胺代谢等代谢通路。该研究利用代谢组学技术分析川牛膝治疗膝骨关节炎的作用机制,为进一步深入研究奠定基础。
Metabolomics based on ultra-high performance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry(UHPLC-Q-TOF-MS) was employed to analyze the effect of Cyathulae Radix on serum and urine in rats with knee osteoarthritis, and to decipher the mechanism of Cyathulae Radix in the treatment of knee osteoarthritis. SD rats were randomized into a normal group, a model group, a positive drug group, and a Cyathulae Radix group. The knee osteoarthritis model was established by injecting 2% pa-pain and 0.03 mol·L-1cysteine, and the serum levels of tumor necrosis factor-α(TNF-α) and matrix metalloproteinase-3(MMP-3) in the model group was measured to preliminarily evaluate the therapeutic effect of Cyathulae Radix on knee osteoarthritis. UHPLC-Q-TOF-MS was employed to establish the metabolic profile of endogenous small molecule metabolites in the four groups. Potential biomarkers were screened out by multivariate analysis methods such as partial least squares discriminant analysis(PLS-DA) and orthogonal partial least squares discriminant analysis(OPLS-DA) in combination with t-test, variable importance for projection(VIP), and fold-change. The related metabolic pathways were enriched with the help of MetaboAnalyst 5.0. The results showed that Cyathulae Radix alleviated the general signs of rats with knee osteoarthritis, and reduced the levels of TNF-α and MMP-3 in the rat serum. Twenty-eight differential metabolites that might be associated with the therapeutic effect of Cyathulae Radix were screened out from serum and urine. They were mainly involved in arginine biosynthesis, nicotinate and nicotinamide metabolism, tricarboxylic acid cycle, alanine, aspartate and glutamate metabolism, riboflavin metabolism, glyoxylate and dicarboxylate metabolism, and pyrimidine metabolism. Through metabonomics analysis, this study predicted the possible mechanism of Cyathulae Radix in the treatment of knee osteoarthritis, which laid a foundation for further research.
作者
刘双晶
李建朋
王彦志
李恺之
朱跃通
冯卫生
LIU Shuang-jing;LI Jian-peng;WANG Yan-zhi;LI Kai-zhi;ZHU Yue-tong;FENG Wei-sheng(School of Pharmucy,Henan University of Chinese Medicine,Zhengzhou 450046,China;Collaboratire Innovation Center for Chinese Medicine and Respiratory Diseases Co-constructed by Henan Protince&Education Ministry of P.R.China,Zhengzhou 450046,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2022年第22期6199-6206,共8页
China Journal of Chinese Materia Medica
基金
河南省重大科技专项(171100310500)。
关键词
川牛膝
膝骨关节炎
代谢组学
潜在生物标志物
Cyathulae Radix
knee osteoarthritis
metabonomics
potential biomarker
