雷公藤红素改善缺血性脑损伤机制的蛋白质组学及代谢组学研究

Proteomic and metabolomic study on the mechanism of celastrol ameliorating ischemic brain injury

关键词:  雷公藤红素  缺血性脑损伤  代谢组  蛋白质组  小鼠

Key words:  celastrol  ischemic brain injury  metabolome  proteome  mice

摘要:

目的采用代谢组学及蛋白质组学技术研究雷公藤红素对缺血性脑损伤保护作用的相关机制。 方法建立ICR小鼠大脑中动脉缺血再灌注模型。将建模成功的小鼠分为对照组及雷公藤红素治疗组，收集脑损伤组织进行代谢组学分析。采用LPS处理BV2小胶质细胞诱导体外脑损伤模型。经雷公藤红素处理后收集细胞样本进行代谢组学及蛋白组学分析。 结果与正常组相比，模型组中有22个显著富集于天冬氨酸盐代谢、苯丙氨酸和酪氨酸代谢、嘌呤代谢、尿素循环等的代谢物发生显著改变；雷公藤红素处理后大多数异常的代谢物得到了回调。LPS处理后，BV2小胶质细胞中有131个生物标志物发生显著改变，这些代谢物显著富集于精氨酸和脯氨酸代谢、苹果酸-天门冬酸穿梭、天冬氨酸代谢等。给药后大多数异常的代谢物得到了回调，其中43个代谢物达显著性差异。蛋白质组共鉴定到7590个蛋白，其中LPS处理诱导393个蛋白发生差异表达。而与LPS组相比，雷公藤红素处理后126个蛋白发生显著差异表达。功能富集分析发现，这些差异蛋白与免疫进程、炎症反应及其相关信号通路。 结论体内及体外脑损伤模型可导致脑组织及细胞参与天冬氨酸代谢、嘌呤代谢等的代谢物以及参与炎症反应的信号通路发生变化，雷公藤红素可回调部分代谢物及差异蛋白的变化。

Abstract:

ObjectiveTo investigate the protective effect and mechanism of celastrol on ischemic brain injury by metabolomics and proteomics analyses. MethodsThe middle cerebral artery ischemia-reperfusion model was established in ICR mice. The mice were divided into control group and celastrol treatment group, and the brain injury tissues were collected for metabolomics analysis. In addition, BV2 mouse microglia was treated with LPS to induce brain injury models in vitro. After treatment with celastrol, cell samples were collected for metabolomics and proteomic analysis. ResultsCompared with the control group, 22 metabolites in the model group were significantly enriched in aspartate metabolism, phenylalanine and tyrosine metabolism, purine metabolism, urea cycle and so on. Most of the abnormal metabolites were reversed after celastrol treatment. After LPS treatment, 131 biomarkers were significantly changed in BV2 microglia, and these metabolites were significantly enriched in arginine and proline metabolism, malate-aspartate shuttle, and aspartic acid metabolism. Most of the abnormal metabolites were reversed after celastrol treatment, and 43 metabolites showed significant differences. A total of 7590 proteins were identified in the proteomic analysis, of which LPS treatment induced differential expression of 393 proteins. Compared with LPS group, 126 proteins were significantly expressed after celastrol treatment. Functional enrichment analysis showed that these differential proteins were associated with immune processes, inflammatory responses and their related signaling pathways. ConclusionIschemic brain injury can lead to the changes of metabolites involved in aspartate and purine metabolism, and the signaling pathways involved in inflammation. Triptolide can regulate the changes of some metabolites and differential proteins.