红景天苷对缺血性脑损伤大鼠的作用

Effect of salidroside on ischemic brain injury in rats

目的基于超高效液相色谱-电喷雾-串联质谱(UPLC-ESI-MS/MS)技术研究红景天苷(salidroside,Sal)的血脑屏障(blood brain barrier,BBB)穿透性,并通过网络药理学、分子对接技术和动物实验探讨Sal治疗缺血性脑卒中(ischemic stroke,IS)的作用靶点及机制。方法利用UPLC-ESI-MS/MS技术研究Sal的BBB穿透性;运用多个数据库预测Sal的作用靶点和IS的疾病靶点,构建PPI网络并筛选核心靶点,进行GO和KEGG富集分析,并通过分子对接技术和动物实验进行验证。结果Sal给药正常大鼠和MCAO大鼠后,血浆和脑组织中均检测到Sal原形和代谢物酪醇;网络药理学共筛选出Sal治疗IS的作用靶点191个,GO功能富集分析主要涉及蛋白水解、细胞迁移的正调控等生物过程;KEGG通路分析提示PI3K-Akt、MAPK、FOXO等信号通路在Sal治疗IS过程中发挥关键作用;分子对接结果显示,Sal与对接的核心靶点均有良好的结合能力;动物实验结果表明,Sal可以明显改善MCAO大鼠的神经功能损伤,明显增加缺血侧Nissl阳性细胞的数量,促进核心靶点VEGF、EGFR和IGF1蛋白的表达,抑制IL-6和MMP9蛋白的表达。结论Sal能够穿透BBB,进入中枢神经系统发挥药理作用;网络药理学预测了Sal治疗IS的核心靶点,包括VEGFA、EGFR、IL-6、MMP9、IGF1、CASP3、ALB、SRC,通过动物实验验证了Sal对部分核心靶点的作用,为今后临床进一步探讨Sal治疗IS的作用机制提供参考依据。

Aim To study the permeability of salidroside(Sal)to the blood brain barrier(BBB)by high-performance liquid chromatography electrospray ionization tandem mass spectrometry(UPLC-ESI-MS-MS),and to explore the target and mechanism of Sal in the treatment of ischemic stroke(IS)by network pharmacology,molecular docking technique and animal experiment.Methods UPLC-ESI-MS/MS was used to study the BBB penetration of Sal.Multiple databases were used to predict the target of Sal and the disease target of IS,GO and KEGG enrichment analysis were performed and verified by molecular docking technique and animal experiments.Results After Sal administration to normal rats and MCAO rats,Sal prototype and the metabolite tyrosol were detected in plasma and brain tissue of rats.A total of 191 targets were identified by network pharmacology,the enrichment analysis of GO mainly involved in the biological processes of proteolysis and positive regulation of cell migration,and the analysis of KEGG pathway suggested that PI3K-Akt,MAPK,FOXO and other signaling pathways played a key role in the treatment of IS by Sal The results of molecular docking showed that Sal had good binding ability with the core target of docking,and the results of animal experiments showed that Sal could significantly improve the neurologic impairment of MCAO rats,the number of Nissl-positive cells in ischemic side significantly increased,and the expression of VEGF,EGFR and IGF1 increased,while the expression of IL-6 and MMP9 was inhibited.Conclusions Sal is able to penetrate the BBB and enter the central nervous system for its pharmacological effects.Network pharmacology predicts the core targets of Sal in the treatment of IS,including VEGFA,EGFR,IL-6,MMP9,IGF1,CASP3,ALB,SRC.The effects of Sal on some core targets can be verified by animal experiments,to provide a reference for further study of the mechanism of Sal in the treatment of IS.