通讯作者: 翁思颖, alicewsy@163.com

DOI：10.12201/bmr.202409.00020

Study on Plasma Metabolomics Analysis of Yanghe-patch in the treatment of impaired glucose tolerance with Yang-deficiency constitution

• 摘要：目的 探讨阳和帖穴位贴敷治疗阳虚体质糖耐量减低人群的疗效及其对血清代谢物的影响。方法 研究于2022年10月至2024年1月在浙江中医药大学附属宁波市中医院体检、门诊、住院就诊人群中纳入63例IGT人群（阳和帖穴位贴敷治疗+生活方式干预组31例，单纯生活方式干预组对照组32例），对比治疗前后2组间血糖、血脂、肿瘤坏死因子α（TNF-α）及阳虚体质评分与红外线热成像区域温度。另外选取29例健康人群作健康阴性对照，运用液相色谱-质谱（LC-MS）技术对3组人群的血清代谢产物进行分析，结合主成分分析（PCA）、潜在结构判别分析（OPLS-DA）等方法分析特征性差异代谢物，运用Metabo Analyst软件进行网络分析以筛选关键代谢通路。结果 阳和贴干预后治疗组空腹血糖、糖化血红蛋白水平下降（P<0.05），甘油三酯、低密度脂蛋白水平较干预前均下降（P<0.05），且均低于对照组（P<0.05），两组总胆固醇水平均无明显变化（P>0.05），TNF-α水平较干预前下降，低于对照组（P<0.05）；治疗组阳虚体质转化分下降（P<0.05），任脉、督脉温度均明显升高（P<0.05），上焦温度下降（P<0.05），中焦、下焦温度均明显升高（P<0.05），对照组治疗前后差异不明显（P>0.05）。代谢组学分析发现阳虚体质糖耐量减低患者血清中的差异表达代谢物主要包括4-乙酰氨基丁酸、3-脱氢胆酸、柠檬酸、胱氨酸等16个，治疗组相较对照组出现差异表达的代谢物包括二氨基庚二酸、乙酰左旋肉碱、甜菜碱等14个。经Metabo Analyst分析，发现糖耐量减低阳虚体质的形成与三羧酸循环、氨基酰tRNA生物合成代谢通路、甘氨酸-丝氨酸-苏氨酸代谢轴（Gly-Ser-Thr）有关，而Gly-Ser-Thr代谢轴与鞘脂代谢则与阳和帖的干预作用相关。结论 阳和帖穴位贴敷可缓解阳虚体质糖耐量减低人群的血清慢性炎症、改善阳虚症状，代谢组学分析发现多种差异性代谢物与其作用机制有关，其中Gly-Ser-Thr代谢轴相关代谢通路被认为是其调护糖耐量减低阳虚体质的关键通路。

  关键词： 糖耐量减低；中医体质学；穴位贴敷；代谢组学

   

  Abstract: Objective To investigate the impact of Yanghe-patch acupoint application on individuals with impaired glucose tolerance (IGT) and its influence on serum metabolites. Methods From Oct.2022 to Jan.2024, a total of 63 patients diagnosed with IGT were enrolled for comprehensive medical care at Ningbo Hospital of TCM affiliated with Zhejiang Chinese Medicine University. The treatment groups consisted of the Yanghe-patch acupoint application therapy combined with lifestyle intervention group (31 cases) and the simple lifestyle intervention group (32 cases).The study compared the changes in blood glucose, blood lipid levels, tumor necrosis factor α(TNF-α), Yang deficiency body scores, and infrared thermal imaging temperature before and after treatment between the two groups. Furthermore, 29 healthy individuals were selected as controls, and their serum metabolites were analyzed using liquid chromatography-mass spectrometry (LC-MS). Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were employed to identify characteristic differential metabolites. Additionally, key metabolic pathways were screened through network analysis using MetaboAnalyst software. Results The treatment group exhibited a significant decrease in fasting blood glucose and glycosylated hemoglobin levels following Yanghe-patch intervention (P<0.05). Furthermore, compared to pre-intervention levels, the treatment group demonstrated reduced triglyceride and low-density lipoprotein levels (P<0.05), which were also lower than those observed in the control group (P<0.05). However, there were no notable changes in total cholesterol levels between the two groups (P>0.05). Additionally, TNF-α levels were significantly lower in the treatment group compared to the control group (P<0.05). In terms of traditional Chinese medicine evaluation parameters, individuals with Yang deficiency physique experienced a decrease at their conversion point after intervention (P<0.05), while Ren pulse and Du pulse temperatures showed a significant increase (P<0.05). Furthermore, Upper-Jiao temperature decreased significantly while Middle-Jiao and Lower-Jiao temperatures increased notably following intervention within the treatment group (P<0.05); there was no significant difference before and after treatment within the control group. Metabolomics analysis revealed that differentially expressed metabolites found in IGT patients with Yang deficiency mainly included 4-acetylaminobutyric acid, 3-dehydrocholic acid, citric acid, cystine; additionally, fourteen metabolites showed differential expression when comparing the treatment group with controls including diaminoheptadecic acid , acetyl-l-carnitine ,and betaine . MetaboAnalyst analysis demonstrated that diabetic yang-deficiency constitution formation correlated with tricarboxylic acid cycle metabolism,glycine-serine-threonine metabolic axis(Gly-Ser-Thr), aminoacyl tRNA bioanabolic pathway; whereas Gly-Ser-Thr metabolic axis,sphingolipid metabolism related to Yang Hetians therapeutic effects. Conclusion The application of Yanghe-patch acupoint has been found to alleviate serum chronic inflammation and improve symptoms of yang-deficiency in IGT patients with a constitution characterized by yang-deficiency. Metabolomics analysis has identified various metabolites associated with their mechanism of action, among which the Gly-Ser-Thr metabolic axis-related pathway is considered as the pivotal pathway for regulating the yang-deficiency constitution in IGT individuals.

  Key words: impaired glucose tolerance；TCM Constitution；Acupoint application；Metabolomics

  提交时间：2024-09-12

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  1	2024-06-29	bmr.202409.00020V1	下载

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