Metabonomic study on the anti-osteoporosis effect of Rhizoma Drynariae and its action mechanism using ultra-performance liquid chromatography-tandem mass spectrometry
Metabonomic study on the anti-osteoporosis effect of Rhizoma Drynariae and its action mechanism using ultra-performance liquid chromatography-tandem mass spectrometry
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DOI：
10.1016/j.jep.2011.11.017
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发表时间：
2012-01-06
影响因子：
5.4
通讯作者：
Xiong, Zhili
中科院分区：
文献类型：
--
作者：
Liu, Xinyu;Zhang, Shangshang;Xiong, Zhili
民族药理学相关性：骨碎补是治疗骨质疏松、骨折、链霉素耳中毒及败血症的有效中药。目前，对RD药理作用机制的研究多集中在细胞和基因水平，对其代谢组学的研究较少。本研究旨在建立正常对照组、模型组和治疗组大鼠的血浆代谢组学，探讨RD的抗骨质疏松作用及其作用机制。模型组和治疗组大鼠注射泼尼松龙12周，同时治疗组大鼠灌胃治疗剂量（10 g/kg，以原料药重量表示）的RD提取物，对照组和模型组大鼠灌胃等量生理盐水。12周后，采集3组大鼠的血浆样品，采用超高效液相色谱-串联质谱（UPLC-MS/MS）分析各组大鼠代谢谱的变化。通过主成分分析（PCA）和偏最小二乘判别分析（PLS-DA）对所得数据集进行分析。使用参比标准品，通过比较其质谱、MS/MS裂解和保留时间，对所有潜在生物标志物进行鉴别。结果：模型组与对照组的PCA评分图有明显的分离趋势，PLS-DA评分图显示RD具有抗骨质疏松作用。在大鼠血浆中鉴定出6种潜在的代谢物生物标志物：溶血磷脂酰胆碱（C16：0 LPC、C18：0 LPC、C18：1 LPC和C18：2 LPC）、色氨酸和苯丙氨酸，它们已被证明与骨质疏松症有关。与对照组相比，模型组各项生物标志物水平均显著升高，而治疗组各项生物标志物水平均接近正常。结论：代谢组学方法可靠地证实了RD的抗骨质疏松作用。RD可能通过干预大鼠体内抗氧化-氧化平衡、胆固醇代谢和苯丙氨酸代谢而预防骨质疏松。(C)2011年由Elsevier爱尔兰有限公司出版。
Ethnopharmacological relevance: Rhizoma Drynariae (RD) is an effectively traditional Chinese medicine which is usually used in treating osteoporosis, bone fracture, streptomycin ototoxicity and hyperlipemia. Up to now, studies on pharmacological mechanism of RD mostly focus on cell and gene level, little is known about its metabonomics study. The aim of this study is to establish the rats plasma metabonomic profiles of control, model and treatment group, then to investigate the anti-osteoporosis effect of RD and its action mechanism.Method: A total of 21 Wistar rats was divided into three groups: control group, model group and treatment group. The model and treatment rats were injected prednisolone for 12 weeks, at the same time the treatment rats were orally administered RD extract at a therapeutic dose (10 g/kg, expressed as the weight of raw material) once daily throughout the experimental period, control group and model group were orally gavaged approximately volume normal saline solution. After 12 weeks, all plasma samples of three groups were collected and their metabolic profiling changes were analyzed by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The resulting dataset was analyzed by principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA). The identification of all potential biomarkers was performed using reference standard by comparing their mass spectra, MS/MS fragmentation and retention time. Furthermore, clinical biochemistry and biomechanics study were also carried out to ensure the success of the osteoporosis model and to investigate the anti-osteoporosis effect of RD.Results: Obvious separation trend between control and model group was found in PCA score plot, the anti-osteoporosis effect of RD can be indicated in PLS-DA score plot among these three groups. Six potential metabolite biomarkers, Lysophosphatidylcholines (C16:0 LPC, C18:0 LPC, C18:1 LPC and C18:2 LPC), tryptophane and phenylalanine, which were proved to be related with osteoporosis, were identified in the rats plasma. Compared with control group, level of all biomarkers increased significantly in model group, while that was much closer to normal in treatment group.Conclusion: The anti-osteoporosis effect of RD has been reliably confirmed by the metabonomics method. The osteoporosis might be prevented by RD via intervening antioxidant-oxidation balance, tryptophane metabolism and phenylalanine metabolism in vivo in rats. (C) 2011 Published by Elsevier Ireland Ltd.