基于代谢组学和网络药理学探讨五色培元固本方改善慢性肾病-蛋白质能量消耗综合征的作用机制

Mechanism of Wuse Peiyuan Guben Decoction in Treating Chronic Kidney Disease-Protein Energy Wasting: Based on Metabolomics and Network Pharmacology

关键词:  慢性肾病  蛋白质能量消耗综合征  五色培元固本方  代谢组学  网络药理学

Key words:  chronic kidney disease  protein-energy wasting  Wuse Peiyuan Guben Decoction  metabolomics  network pharmacology

摘要:

目的 探讨五色培元固本方治疗慢性肾病-蛋白质能量消耗综合征(chronic kidney disease-protein energy wasting，CKD-PEW)的潜在作用机制。方法 采用5/6肾切术搭配低蛋白饮食建立大鼠CKD-PEW模型，通过广泛靶向代谢组学技术对大鼠血清进行代谢组学分析，得到差异代谢物及对应靶点；通过网络药理学预测五色培元固本方改善CKD-PEW的化学成分、靶点及信号通路。整合分析代谢组学及网络药理学共有靶点，并通过共有靶点构建“成分-靶点-代谢物-通路”关系网络图，揭示五色培元固本方治疗CKD-PEW的主要化学成分和代谢机制。结果 血清代谢组学发现在空白组和模型组间存在123种差异代谢物，模型组与阳性对照开同组以及五色培元固本低、中、高剂量组比较，分别存在50，37，62和114种差异代谢物，经Metabo Analysis进行代谢通路分析富集在代谢通路上的潜在生物标志物，空白组和模型组间存在25种生物标志物，模型组与阳性对照开同组、五色培元固本低、中、高剂量组间分别存在2，2，5和24种生物标志物；空白组和模型组间富集到 21 条通路及194个靶点，模型组与阳性对照开同组、五色培元固本低、中、高剂量组间分别富集到2 条通路14个靶点、2条通路14个靶点、5条通路 38个靶点、24条通路52个靶点。网络药理学分析五色培元固本方与CKD-PEW疾病得到65种活性成分及123个关键靶点。代谢组学及网络药理学整合分析模型组与药物高剂量组间得到髓过氧化物酶、尿苷二磷酸葡萄糖醛基转移酶1A1、酪氨酸酶、氧磷酶14个共同靶点，结果表明这4个靶点可能受到五色培元固本方中β-谷甾醇、槲皮素、柚皮素以及红景天苷4种成分调控，从而干预2-羟基丁酸、对苯二酚、2-苯乙酰胺、多巴醌4种生物标志物，最终影响苯丙氨酸代谢、丙酸代谢、酪氨酸代谢3条代谢通路，达到治疗CKD-PEW的目的。结论 CKD-PEW病因与多靶点、多途径有关，表明五色培元固本方能够通过多成分、多靶点及多代谢通路改善CKD-PEW的作用机制，也为临床治疗CKD-PEW提供了研究基础。

Abstract:

OBJECTIVE To explore the potential mechanism of Wuse Peiyuan Guben Decoction in treating chronic kidney disease-protein energy wasting(CKD-PEW). METHODS The CKD-PEW model on rats was established by using a 5/6 nephrectomy and low protein diet. Widely targeted metabolomics technology was employed for the serum metabolomics analysis on rats to identify differential metabolites and their corresponding targets, whereas network pharmacology method was utilized to predict the key chemical compositions, targets, and metabolic pathways of Wuse Peiyuan Guben Decoction in treating CKD-PEW. Furthermore, the common targets discovered using the integrated serum metabolomics and network pharmacology analysis were leveraged to construct the component-target-metabolite-pathway network, which presented the main effective chemical compositions and metabolic mechanisms of Wuse Peiyuan Guben Decoction in treating CKD-PEW. RESULTS There were 123 different metabolites between the blank group and the model group, and 50, 37, 62, and 114 different metabolites in the model group compared with the positive control compound α-ketoacid tablets, low, medium, and high dosage Wuse Peiyuan Guben Decoction groups, respectively. There were 25 potential biomarkers enriched in the metabolic pathway between the blank group and the model group by Metabo Analysis, and 2, 2, 5 and 24 potential biomarkers in the model group and the positive control compound α-ketoacid tablets, low, medium and high dosage Wuse Peiyuan Guben Decoction groups, respectively. There were 21 enriched metabolic pathways with 194 targets between the blank group and the model group, and 2 pathways with 14 targets, 2 pathways with 14 targets, 5 pathways with 38 targets, and 24 pathways with 52 targets between the model group and positive control compound α-ketoacid tablets, low, medium and high dose groups, respectively. There were 65 active pharmaceutical ingredients and 123 important targets identified by the network pharmacology analysis. The integrated serum metabolomics and network pharmacology analysis results presented that four common targets: myeloperoxidase, UDP-glucuronosyltransferases, tyrosinase, and paraoxonase-1, were regulated by the four compositions, beta-sitosterol, quercetin, naringenin, salidroside in Wuse Peiyuan Guben Decoction to impact the key biomarkers, 2-hydroxybutanoic acid, hydroquinone, 2-phenylacetamide, and dopaquinone, thereby to affect the metabolic pathways of phenylalanine, propanoate, and tyrosine to finally treat CKD-PEW. CONCLUSION The pathogenesis of CKD-PEW is associated with multiple targets and metabolic pathways, and this study has elaborated on the key effective chemical compositions and metabolic mechanisms of Wuse Peiyuan Guben Decoction in treating CKD-PEW, hopefully providing a solid theoretical and experimental foundation for medical treatment of CKD-PEW.