【作者】 魏岩；

【导师】 黄永生；

【作者基本信息】 长春中医药大学 ， 中医内科学， 2011， 博士

【摘要】 研究目的本研究利用代谢组学技术对冠心病先天伏寒证患者给药前后、冠心病心绞痛非先天伏寒证患者及健康人的血液进行对比分析,拟通过分析患者血液代谢物质的变化探索病证模式背后的生物标记物,为冠心病心绞痛先天伏寒证本质的系统研究提供科学依据。建立病证结合的代谢组学研究技术平台,为从代谢物水平探讨中医证本质提供思路和方法。研究方法1.冠心病心绞痛先天伏寒证、非先天伏寒证患者的血清代谢组学研究:对10例冠心病心绞痛先天伏寒证、10例冠心病心绞痛非先天伏寒证患者的血清样本进行氢核磁共振检测(1H-Nuclear Magnetic Resonance,1H-NMR)。通过偏最小二乘法判别分析方法研究冠心病心绞痛先天伏寒证、非先天伏寒证患者之间血清代谢产物谱的差异。2.冠心病先天伏寒证患者给药前后的血清代谢组学研究:对10例临床疗效确切的冠心病先天伏寒证患者进行给药前后血清样本的1H-NMR检测,同时10例健康人作为为正常对照组。通过偏最小二乘判别分析方法研究各组之间的血清代谢产物谱差异。结果1.冠心病心绞痛先天伏寒证、非先天伏寒证患者的的血清代谢物可以被分开,两组存在明显差异的代谢产物:丙氨酸、低密度脂蛋白/极低密度脂蛋白、脂类化合物、谷氨酸、谷氨酰胺、α-葡萄糖、高密度脂蛋白。2.冠心病心绞痛先天伏寒证患者与健康人血清代谢物存在明显差异。与正常组相比,冠心病先天伏寒证组中丙氨酸、乳酸、低密度脂蛋白/极低密度脂蛋白、谷氨酸、谷氨酰胺、α-葡萄糖、β-葡萄糖、N-乙酰糖蛋白、脂类化合物等含量较高,有统计学意义(P<0.05);而高密度脂蛋白含量则较低,有统计学意义(P<0.05)。3.伏寒颗粒治疗前后患者血清代谢谱的比较:与治疗前组相比,冠心病先天伏寒证患者治疗后组中高密度脂蛋白、丙氨酸含量较高,其中高密度脂蛋白与治疗前组相比有统计学意义(P<0.05),而丙氨酸与治疗前组相比无统计学意义(P>0.05);而乳酸、低密度脂蛋白/极低密度脂蛋白、谷氨酸、谷氨酰胺、α-葡萄糖、β-葡萄糖、脂类化合物、N-乙酰糖蛋白等物质含量较低,其中乳酸、低密度脂蛋白/极低密度脂蛋白、谷氨酸、谷氨酰胺、α-葡萄糖、β-葡萄糖、脂类化合物与治疗前组相比有统计学意义(P<0.05),而N-乙酰糖蛋白与治疗前组相比无统计学意义(P>0.05)。证明用伏寒颗粒干预后的代谢物(如能量代谢异常方面的指标等)可降低或者趋近于正常组,这些物质的改变途径可能是伏寒颗粒的作用机制,有待进一步研究。结论各组血液样本的1H-NMR代谢谱存在差异,能从代谢组学分析中找出特异的标志性代谢产物。利用代谢组学方法能区分不同证型的冠心病心绞痛患者的血清1H-NMR代谢谱,能确定先天伏寒证与非先天伏寒证之间存在明显差异的代谢产物。冠心病先天伏寒证存在着能量代谢、糖脂代谢、免疫调节等多种代谢通路的异常,高密度脂蛋白、丙氨酸、乳酸、低密度脂蛋白/极低密度脂蛋白、谷氨酸、谷氨酰胺、α-葡萄糖、β-葡萄糖、N-乙酰糖蛋白、脂类化合物有可能成为冠心病先天伏寒证的潜在生物标记物。祖国医学和代谢组学均是对影响机体整体状况的多因素的研究,对机体某一病理变化的全面情况进行探究。二者之间理论上的关联以及本研究的结果,为中医辨证分型客观化研究提供了新思路,为揭示中医学证的本质提供了新方法。这为我们进一步研究证候本质提供了思路。更多还原

【Abstract】 Purposewe used metabonomics technology for comparison of blood among angina pectoris patients with congenital latent cold syndrome and simple angina pectoris patients as well as healthy people. We tried to explore biomarkers underlying patterns of disease and syndrome by analyzing changes of blood and to provide scientific evidence for angina pectoris with congenital latent cold syndrome study. By establishment of disease and syndrome combing metabonomics research technology, we provide ideas and methods to explore the nature of traditional Chinese medicine syndromes.Method1. Plasma metabonomics study for angina pectoris patients with congenital latent cold syndrome and simple angina pectoris patients: plasma samples from 10 cases with congenital latent cold syndrome and 10 cases with simple angina pectoris were detected by nuclear magnetic resonance (1H-NMR). Difference of plasma metabolites between patients with congenital latent cold syndrome and simple angina pectoris patients was studied by discriminant analysis (PLS-DA) method.2. Plasma metabonomics study on coronary heart disease with congenital latent cold syndrome before and after treatment: plasma sample from 10 cases with congenital latent cold syndrome was detected by nuclear magnetic resonance (1H-NMR) and the sample of 10 healthy cases was compared as a normal control group. Difference of plasma metabolites between two groups was analyzed by discriminant analysis (PLS-DA) method.Result1. Plasma metabolites of angina pectoris patients with congenital latent cold syndrome and those without congenital latent cold syndrome varied. There were significantly different metabolites: Alanine, LDL, VLDL, Lipid, GAA, Glutamine, GLU, HDL.2. There were different plasma metabolites between angina pectoris patients with congenital latent cold syndrome and healthy people. Compared with healthy people, Alanine, Lactate, LDL, VLDL, GAA, Glutamine,α-GLU,β-GLU,NAC, Lipid increased in patients with congenital latent cold syndrome. While HDL decreased considerably. There was statistic difference.3. Comparison of plasma metabolites spectrum before and after treatment by Fu Han Granule. HDL and Alanine increased in congenital latent cold syndrome patients after treatment. There was statistic difference in HDL. While Lactate, LDL, VLDL, GAA, Glutamine,α--GLU,β-GLU, Lipid and NAC decre- ased. There was significant difference in those except NAC. It shows that metabolites after treatment given by Fu Han Granule could decrease or be close to normal range. It may be the action mechanism of Fu Han Granule, which requires further research.Conclusion:Metabolic products among these samples have significant difference, and characteristic products can be detected by metabonomics analysis. By metabonomics technology we can distinguish metabolic products from different syndromes. There is abnormal metabolism of energy, glucose and lipid in patients with congenital latent cold syndrome. HDL, Alanine, Lactate, LDL, VL-DL, GAA, Glutamine,α-GLU,β-GLU, NAC and Lipid may be potential biomar-kers of coronary heart disease with congenital latent cold syndrome. Both traditional Chinese medicine and metabonomics focus on study of multi-factors on the whole. Integration of theories and result of research provide new ideas and methods to explore syndrome differentiation and essence of traditional Chinese medicine.更多还原