【作者】 范群丽；

【导师】 陈晓虎；

【作者基本信息】 南京中医药大学 ， 中西医结合临床， 2011， 博士

【摘要】 目的：①研究和分析高血压肝火亢盛证及痰湿壅盛证患者血浆内源性小分子代谢物的差异,鉴定与证型相关的生物标志物群；②研究和分析高血压人群与正常组的血浆代谢谱,分析高血压人群体内的代谢组学异常；③探讨个体对降压药敏感程度与高血压中医证型的相关性,构筑个体对降压药敏感性——代谢组——中医证型的桥梁。方法：高血压组119例,正常对照组(K组)25例。高血压组分为氨氯地平组(A组)59例,厄贝沙坦组(B组)60例。氨氯地平组分为药物敏感组(E+M)29例,不敏感组(F+N)30例；按中医辨证分型氨氯地平敏感组分为肝火亢盛证组(M组)14例,痰湿壅盛证组(E组)15例；氨氯地平不敏感组分为肝火亢盛证组(N组)15例,痰湿壅盛证组(F组)15例。厄贝沙坦组分为药物敏感组(G+L)30例,不敏感组(H+P)30例；按中医辨证分型厄贝沙坦敏感组分为肝火亢盛证组(L组)13例,痰湿壅盛证组(G组)17例；厄贝沙坦不敏感组分为肝火亢盛证组(P组)16例,痰湿壅盛证组(H组)14例。采用基于气相色谱-飞行时间质谱(GC-TOF/MS)代谢组学方法测定各组人群血浆代谢谱,应用偏最小二乘-判别分析(PLS-DA)比较氨氯地平组内肝火亢盛组与痰湿壅盛组、厄贝沙坦组内肝火亢盛组与痰湿壅盛组、氨氯地平组与对照组、厄贝沙坦组与对照组、氨氯地平药物敏感组与不敏感组、厄贝沙坦药物敏感组与不敏感组,得到各样本的主成分得分图(scores plot),分析和鉴定组间异同内源性化合物。结果：①在氨氯地平组和厄贝沙坦组中,采用GC/MS的代谢组学方法均能区分肝火亢盛证型和痰湿壅盛证型的代谢谱,与肝火亢盛证相关的生物标志物群可能是葡萄糖、木糖、乳酸、缬氨酸、甘氨酸的升高以及α-亚麻酸、甘油酸的降低;与痰湿壅盛证相关的生物标志物群可能是α-亚麻酸、甘油酸的降低以及缬氨酸、甘氨酸、羟脯氨酸、S-甲基-L-半胱氨酸的升高；②氨氯地平药物敏感组与不敏感组表现为不同的代谢特征,在氨氯地平敏感组有10种化合物均低于不敏感组(P<0.05),氨氯地平敏感组内代谢谱表现为糖代谢及部分氨基酸的水平较低；③厄贝沙坦药物敏感组与不敏感组表现为不同的代谢特征,在厄贝沙坦敏感组与不敏感组中有显著差异的化合物有8种(P<0.05),厄贝沙坦敏感组代谢谱表现为氨基酸水平较高,而脂类物质较低；④氨氯地平敏感组与痰湿壅盛证组表现为相似的代谢谱——糖类(葡萄糖)及部分氨基酸(缬氨酸、羟脯氨酸)的水平较低,厄贝沙坦敏感组和肝火亢盛证表现为相似的代谢谱——部分氨基酸(缬氨酸、甘氨酸)及脂类的水平较低。结论：①高血压肝火亢盛证和痰湿壅盛证的血浆代谢谱变化为中医辨证分型提供了部分客观化的科学依据；②高血压人群存在糖、脂、氨基酸代谢异常,代谢组学方法具有研究高血压病理的优势；③降压药物的敏感性与中医证型在代谢学上的联系,对临床使用个体化的思路来选择降压药物提供帮助,并为中西医结合治疗高血压构筑桥梁。更多还原

【Abstract】 Objectives:①Research and analyze the difference of endogenous plasma small molecule metabolites between overabundant liver-fire syndrome and phlegm-dampness accumulation syndrome, and identify syndrome-related biomarker cluster.②Research and analyze plasma metabolomic profiling between hypertension group and normal group, analyze abnormality of metabolomics in people with hypertension③Explore the correlation between individual sensitivity to antihypertensive drugs and TCM syndrome types to establish a bridge between these two and metabolome.Methods:119 patients with hypertension, in addition to normal control group (K group) with 25 cases. Hypertension group is divided into amlodipine group (A group) with 59 cases, irbesartan group (B group) with 60 cases. Amlodipine group is divided into drug-sensitive group (E+M) with 29 cases, and non-sensitive group (F+N) with 30 cases. According to TCM syndrome differentiation, amlodipine sensitive group is then divided into abundant liver-fire syndrome group (M group) with 14 cases and phlegm-dampness accumulation syndrome group (E group) with 15 cases, while amlodipine non-sensitive group is divided into abundant liver-fire syndrome group (N group) with 15 cases and phlegm-dampness accumulation syndrome group (F group) with 15 cases. Irbesartan group is separated into drug-sensitive group (G+L) with 30 cases as well as non-sensitive group (H+P) with 30 cases. The former group is then sub-divided into abundant liver-fire syndrome group (L group) with 13 cases and phlegm dampness accumulation syndrome group (G group) with 17 cases, while the later group is divided into hyperactive liver-fire syndrome group (P group) with 16 cases and phlegm dampness accumulation syndrome group (H group) with 14 cases according to TCM syndrome differentiation. The next step is to apply gas chromatography - time of flight mass spectrometry (GC-TOF/MS) to test plasma metabolomic profiling in all groups, and then to compare hyperactive liver-fire syndrome group to phlegm dampness accumulation syndrome group in amlodipine and irbesartan group, amlodipine group to control group, irbesartan group to control group, amlodipine sensitive group to non-sensitive group, irbesartan sensitive group to non-sensitive group, by way of Partial Least Squares-Discriminative Analysis (PLS-DA), so as to obtain each sample’s score plot of principal components as well as to analyze and identify similarities and differences of endogenous compounds between groups.Results:①In amlodipine group and irbesartan group, the metabolomic profiling of abundant liver-fire syndrome and phlegm dampness accumulation syndrome can be distinguished by gas chromatography - time of flight mass spectrometry (GC-TOF/MS) metabolomic method, Biomarker cluster associated with abundant liver-fire syndrome group may be higher glucose, xylose associated with glucose metabolism, lactic acid, valine and glycine belonging to amino acid but lower a-linolenic acid and glycerol acid; while biomarker cluster of phlegm dampness accumulation syndrome group may be lower a-linolenic acid and glycerol belonging to lipid but higher valine, glycine、hydroxyproline and S-methyl-L-cysteine.②Amlodipine sensitive group and non-sensitive group have different metabolomic characteristic,10 compounds in amlodipine sensitive group are lower than non-sensitive group (P<0.05). and metabolomic profiling in amlodipine sensitive group exhibits a lower level of glucose metabolism and part of the amino acids.③Irbesartan sensitive group and non-sensitive group also showing different metabolomic characteristic, there are 8 significant different compounds in irbesartan sensitive group and non-sensitive group (P<0.05), the metabolomic profiling of irbesartan sensitive group shows higher amino acids but lower lipide.④Amlodipine sensitive group and phlegm dampness accumulation syndrome group show similar metabolomic profiling——lower level of sugar (glucose) and some amino acids (valine, hydroxyproline), and the same goes with irbesartan sensitive group and abundant liver-fire syndrome group——lower level of amino acids (valine, glycine) and level of lipids.Conclusions:①Plasma metabolomic profiling of abundant liver-fire syndrome and phlegm-dampness accumulation syndrome of hypertension can supply some objective evidence for syndrome differentiation.(2)Hypertension patients have abnormal metabolism of sugar, fat, amino acid during metabolism, so metabolomics takes advantages in researching the pathology of hypertention.③The connection between the sensitivity of anti-hypertensive drug and TCM syndrome types in metabolomics, which can help us use individual thinking to choose antihypertension drugs, and established a bridge between integrative treatment of hypertension.更多还原