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为了维医证候研究逐步走向标准化和客观化,采用核磁共振(nuclear magnetic resonance,NMR)为基础的代谢组学技术对哮喘异常黑胆质证动物模型体内代谢变化信息与临床患者进行对比研究,揭示两者之间的异同点。分别选择哮喘异常黑胆质证大鼠模型和临床患者作为研究对象,采集血浆样本,通过NMR测试其氢谱,并采用正交偏最小二乘判别分析法(orthogonal partial least-squares discriminant analysis,OPLS-DA)进行多变量分析。结果显示,与健康志愿者相比,临床患者血浆中亮氨酸、异亮氨酸、缬氨酸和丙氨酸含量明显降低;与脂代谢相关的极低密度脂蛋白(VLDL)和不饱和脂类在患者体内明显降低,而乙酸、丙酮、乙酰乙酸和肉碱含量明显升高;谷氨酰胺在患者体内明显的升高,而肌酸、肌酸酐、鲨肌醇和乳酸明显下降。对动物模型而言,健康对照组与2种模型组比较,模型组大鼠血浆中与脂代谢相关的代谢物以及乳酸、肉碱和谷氨酰胺含量的变化趋势与临床患者一致,但氨基酸含量的变化相反;异常黒胆质证模型大鼠与哮喘异常黒胆质证模型大鼠相比较,只有丙氨酸和肌酸酐含量在哮喘异常黒胆质证模型大鼠血浆中明显比前者低。研究表明,虽然氨基酸的代谢变化在模型大鼠和临床患者体内有所不同,但是与脂代谢相关的代谢物含量的变化趋势一致。氨基酸代谢的不同可能与模型制作过程中环境的剧烈变化有关。脂代谢相关的代谢物可作为临床患者与动物模型共同的异常黑胆质证代谢标志物。
In order to standardize and objectify the study of traditional Chinese medicine (TCM) syndromes, the metabonomics based on nuclear magnetic resonance (NMR) was used to compare the in vivo metabolic changes of asthmatic models and clinical patients Similarities and differences between the two points. Plasma samples were collected from the rat model and clinical patients with asthma abnormal savda syndrome, and their hydrogen spectra were determined by NMR. Orthogonal partial least-squares discriminant analysis (OPLS- DA) for multivariate analysis. The results showed that the content of leucine, isoleucine, valine and alanine in the plasma of clinical patients was significantly lower than that of healthy volunteers. The levels of VLDL and unsaturated Lipid levels were significantly reduced in patients, while acetate, acetone, acetoacetate and carnitine levels were significantly increased; glutamine was significantly elevated in patients and creatine, creatinine, scyllo-inositol and lactate were significantly decreased. In the animal model, compared with the two model groups, the changes of the lipid metabolism-related metabolites, lactic acid, carnitine and glutamine in the plasma of the model group were the same as those in the clinical model, but the amino acid content In contrast, only the levels of alanine and creatinine were significantly lower in the asthmatic model rats with asthma than in the asthmatic model rats with abnormal bradycardia syndrome. Studies have shown that although the metabolic changes of amino acids in the model rats and clinical patients are different, but the metabolic changes associated with lipid metabolism trends. The differences in amino acid metabolism may be related to the drastic changes in the environment during the model making process. Metabolites associated with lipid metabolism can serve as metabolic markers of abnormal savda syndrome in clinical patients and animal models.