背景:黄芪和红花(AC)的组合在脑缺血/再灌注损伤(CIRI)中具有显着的治疗作用。了解脑微区域的代谢特征以及组织和体循环的紊乱对于阐明CIRI的机制和AC的治疗益处至关重要。然而,复杂大脑结构的原位代谢调节尚未得到充分研究,AC的治疗机制需要立即澄清。
目的:本研究旨在揭示CIRI在系统和微观区域水平的特定代谢重编程,确定关键的代谢途径和代谢物,阐明AC的治疗机制。
方法:气流辅助解吸电喷雾电离质谱成像(AFADESI-MSI),一种新开发的技术,用于研究脑微区域的代谢物。苏木精-伊红,Nissl,和免疫荧光染色进行可视化与空间代谢相关的微观变化。对血清进行了全面的代谢组学研究,脑组织,和微观区域,连同神经学评估,脑梗死测量,和EvansBlue实验,评估AC治疗对CIRI的全身和局部代谢作用。
结果:AC显著减轻了神经损伤,梗死面积最小化,并修复了CIRI大鼠的血脑屏障损伤。AFADESI-MSI证明CIRI引起的代谢失衡主要发生在大脑皮层,海马体,尾状壳核,丘脑,小脑皮质,和纤维束区域。在这些地区观察到16种代谢物的显著变化,对应于神经元损伤,胶质细胞活化,和神经修复。血清中的20种代谢物和脑组织中的4种代谢物与假手术组的差异显着。综合代谢组学分析表明血清之间存在密切的关系,组织,和微区域代谢。CIRI诱导的全身和局部代谢紊乱涉及14种代谢途径。AC通过逆转各种代谢失衡赋予CIRI的治疗益处。
结论:AFADESI-MSI可有效观察大脑微区代谢。全面的代谢组学分析揭示了CIRI中特定代谢重编程和AC治疗影响的详细见解。AC作为现有CIRI治疗的辅助治疗显示出显著的临床潜力。
BACKGROUND: The combination of Astragalus membranaceus and Carthamus tinctorius (AC) exhibits significant therapeutic effects in cerebral ischemia/reperfusion injury (CIRI). Understanding the metabolic characteristics of brain microregions and disturbances in tissues and systemic circulation is crucial for elucidating the mechanisms of CIRI and the therapeutic benefits of AC. However, in situ metabolic regulation of the complex brain structure has not been adequately studied, and the therapeutic mechanism of AC requires immediate clarification.
OBJECTIVE: The present study aimed to unveil the specific metabolic reprogramming of CIRI at systemic and microregional levels, identify key metabolic pathways and metabolites, and elucidate the therapeutic mechanisms of AC.
METHODS: Air flow-assisted desorption electrospray ionization mass spectrometry imaging (AFADESI-MSI), a newly developed technique, was used to investigate metabolites in brain microregions. Hematoxylin-eosin, Nissl, and immunofluorescence staining were performed to visualize the microscopic changes associated with spatial metabolism. A comprehensive metabolomics study was conducted on serum, brain tissue, and microregions, along with neurological assessments, cerebral infarction measurements, and Evans blue experiments, to assess the systemic and local metabolic effects of AC treatment for CIRI.
RESULTS: AC significantly reduced neurological damage, minimized infarct size, and repaired blood-brain barrier damage in CIRI rats. AFADESI-MSI demonstrated that the metabolic imbalance caused by CIRI primarily occurs in the cerebral cortex, hippocampus, caudate putamen, thalamus, cerebellar cortex, and fiber tract regions. Significant changes in 16 metabolites were observed in these regions, corresponding to neuron damage, glial cell activation, and neural repair. 20 metabolites from serum and 4 from brain tissue varied significantly with the sham group. Comprehensive metabolomics analysis indicated a close relationship among serum, tissue, and microregional metabolism. CIRI-induced systemic and localized metabolic disorders involve 14 metabolic pathways. AC conferred therapeutic benefits in CIRI by reversing various metabolic imbalances.
CONCLUSIONS: AFADESI-MSI efficiently visualized brain microregion metabolism. Comprehensive metabolomics analysis revealed detailed insights into the specific metabolic reprogramming in CIRI and the therapeutic impacts of AC. AC demonstrated significant clinical potential as an adjunct therapy to existing CIRI treatments.