雪菊醇提物对糖尿病气阴两虚证候细胞代谢物的影响

Effect of ethanol extract of Coreopsis tinctoria on cell metabolites of Qi-yin deficiency syndrome in diabetes

中文关键词:  糖尿病体外模型  雪菊醇提物  代谢组学  差异性代谢物

英文关键词:in vitro model of diabetes  ethanol extract of Coreopsis tinctoria  metabolomics  differential metabolites

中文摘要:

      目的 研究雪菊醇提物对体外诱导气阴两虚证候细胞代谢物的影响。方法 噻唑蓝实验检测细胞毒性; 采用核磁共振(nuclear magnetic resonance, NMR)代谢组学方法检测证候细胞裂解液和细胞培养上清液中内源性代谢物变化; 用多元统计分析差异性代谢物与通路的变化。结果 建立糖尿病气阴两虚证候细胞模型, 雪菊醇提物干预后改善了细胞生长活力。代谢组学结果显示: 本实验的所有样本点都分布在95%的置信区间内(即散点图的4个区域中), 聚类分布显著, 且可信度较高。代谢物分析发现与正常组相比, 证候细胞模型组的异亮氨酸、亮氨酸、α-葡萄糖和琥珀酸的含量显著上升(P＜0.05或P＜0.01), 果糖含量显著下调(P＜0.05); 与证候细胞模型组相比, 雪菊醇提物组的脂肪酸、脯氨酸、甘油磷酯酰胆碱、抗坏血酸、α-葡萄糖含量显著下调(P＜0.05或P＜0.01)。差异性代谢物主要涉及氨基酸代谢途径、丙酮酸代谢途径及乙醛酸盐和二羧酸盐代谢途径。结论 采用代谢组学技术进行全面、整体性的分析雪菊醇提物对糖尿病气阴两虚证细胞内代谢物的影响, 为雪菊降低不同证候糖尿病的药用性功效提供基础理论依据。

英文摘要:

      Objective To study the effect of ethanol extract of Coreopsis tinctoria on metabolites of Qi-yin deficiency syndrome cells induced in vitro. Methods Methyl thiazolyl tetrazolium experiment was used to detect cytotoxicity. 1H-NMR metabolomics method was used to detect changes in endogenous metabolites in syndrome cell lysate and cell culture supernatant. Multivariate statistical analysis was used to analyze changes in differential metabolites and pathways. Results Cell model of diabetic syndrome of qi and yin deficiency was established. After the intervention of ethanol extract of Coreopsis tinctoria, cell growth activity was improved. Metabolomics results showed that all the sample points in this experiment were distributed within 95% confidence intervals (that is, in the 4 areas of the scatter plot), the clustering distribution was significant, and the credibility was high. Metabolite analysis found that compared with the normal group, the content of isoleucine, leucine, α-glucose and succinic acid in the syndrome cell model group was significantly increased (P<0.05 or P<0.01), and the content of fructose was significantly reduced (P＜0.05); compared with the syndrome cell model group, the content of fatty acids, proline, glycerophosphoryl choline, ascorbic acid and α-glucose in the ethanol extract of Coreopsis tinctoria group was significantly lowered (P<0.05 or P<0.01). Differential metabolites mainly involved in amino acid metabolic pathways, pyruvate metabolic pathways, and glyoxylate and dicarboxylate metabolic pathways. Conclusion Using metabolomics technique to analyze comprehensively and holistically the effect of ethanol extract of Coreopsis tinctoria on the intracellular metabolites of diabetic qi and yin deficiency syndromes, which provides a basic theoretical basis for the efficacy of ethanol extract of Coreopsis tinctoria in reducing the medicinal properties of diabetes with different syndromes.