黄连解毒汤干预胃火炽盛证引起的口腔疾病患者尿液代谢组学研究

Urinary Metabolomics Study of Huanglian Jiedu Decoction in Patients with Oral Oiseases Caused by Syndrome of Excessive Heat in Stomach Fire

[目的]运用代谢组学方法,研究胃火炽盛证引起的口腔疾病患者的尿液代谢物变化及黄连解毒汤对其干预作用,探索胃火炽盛证引起的口腔疾病患者尿液中的潜在标志物。[方法]按照诊断标准、纳入标准、排除标准等纳入胃火炽盛证引起的口腔疾病患者58例作为口腔疾病患者组,所有患者服用黄连解毒汤5d后为黄连解毒汤干预组,另外纳入健康对照组10例。利用超高效液相色谱-静电场轨道阱质谱联用(ultra performance liquidchromatography coupled with orbitrap tandem mass spectrometry, UHPLC-Orbitrap MS)法对口腔疾病患者黄连解毒汤干预前和干预后,以及健康对照组的尿液进行分析,采用主成分分析(principal component analysis, PCA)和正交偏最小二乘法-判别分析(orthogonal partial least squares-discriminantanalysis, OPLS-DA)处理数据,区分代谢轮廓并寻找可能的生物标记物。[结果]健康对照组和口腔疾病患者组初步鉴定38个差异代谢物,口腔疾病患者组黄连解毒汤干预前和干预后初步鉴定出36个差异代谢物。3组中鉴定出α-N-苯乙酰基-L-谷氨酰胺等10种化合物可能与黄连解毒汤干预胃火炽盛证引起的口腔疾病的作用机制和代谢途径有关,分别为癸酰肉碱、α-N-苯乙酰基-L-谷氨酰胺、反-2,顺-4-癸二烯酰肉毒碱、异亮羟基脯氨酸、3-氨基己酸、10,20-二羟基二十碳酸、菊蒿醇B、Sterebin A、亮菌素和L-谷氨酰胺。推测黄连解毒汤治疗胃火炽盛证口腔疾病主要涉及的代谢通路为三聚氰胺、天冬氨酸和谷氨酸代谢,D-谷氨酰胺和D-谷氨酸代谢等。[结论]本研究首次从代谢组学角度研究了健康对照组和胃火炽盛证口腔疾病患者组,以及胃火炽盛证口腔疾病患者黄连解毒汤干预前和干预后尿液中代谢模式差异,证实黄连解毒汤可以显著修复胃火炽盛证引起的口腔疾病患者尿液中的内源性小分子代谢紊乱。

[Objective] To study the changes of urine metabolites in patients with dental diseases caused by syndrome of excessive heat in stomach fire and the intervention effect of Huanglian Jiedu Decoction on them using urinary metabonomics. Meanwhile, the potential markers in urine of patients with dental diseases caused by syndrome of excessive heat in stomach fire were explored. [Methods] According to the diagnostic criteria, inclusion criteria,exclusion criteria, etc, 58 cases of dental diseases caused by syndrome of excessive heat in stomach fire were included as oral disease group, the 58 cases acted as Huanglian Jiedu Decoction intervention group after treated by Huanglian Jiedu Decoction for 5 days and 10 subjects in healthy control group.Ultra performance liquid chromatography coupled with orbitrap tandem mass spectrometry(UHPLC-Orbitrap MS) method was used to analyze the urine of healthy control group, oral disease group before and after intervention of Huanglian Jiedu Decoction. The metabolic profile was distinguished by principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA). Then the potential biomarkers were searched. [Results]Thirty eight different metabolites were identified between healthy control group and oral diseases group. Thirty six different metabolites were identified in patients of oral disease group before and after intervention of Huanglian Jiedu Decoction. In the three groups, ten potential biomarkers were identified to be related to the mechanism and metabolic pathway of Huanglian Jiedu Decoction in interfering with the syndrome of excessive heat in stomach fire,including decanoylcarnitine, alpha-N-phenylacetyl-L-glutamine, 2-trans,4-cis-decadienoylcarnitine, isoleucyl-hydroxyproline, 3-Aminocaproic acid,10,20-dihydroxyeicosanoic acid, tanacetol B, Sterebin A, armillarin and L-glutamine. The intervention of Huanglian Jiedu Decoction mainly affected alamine, aspartate and glutamate metabolism, D-glutamine and D-glutamate metabolism. [Conclusion] For the first time, this study studied the differences in metabolic patterns between the healthy control group and the patients with oral diseases caused by syndrome of excessive heat in stomach fire, the differences in the metabolism patterns in the urine of the patients with oral disease caused by syndrome of excessive heat in stomach fire and Huanglian Jiedu Decoction intervention group from the perspective of metabolomics. Huanglian Jiedu Decoction can significantly repair endogenous small molecular metabolic disorders in urine samples of patients with oral diseases caused by syndrome of excessive heat in stomach fire.