Kuppuswamy Sivaraman, Watson Noah J, Ledford William Luke, Pavri Blake A, Zhi Wenbo, Gbadebo Mary, Bonsack Frederick, Xu Hongyan, Sukumari-Ramesh Sangeetha
Department of Pharmacology and Toxicology, Medical College of Georgia, Augusta University, 1120, 15th Street, CB3515, Augusta, GA 30912, United States of America.
Department of Pharmacology and Toxicology, Medical College of Georgia, Augusta University, 1120, 15th Street, CB3515, Augusta, GA 30912, United States of America.
Neurobiol Dis. 2025 Aug;212:106936. doi: 10.1016/j.nbd.2025.106936. Epub 2025 May 2.
Aging is an independent predictor of adverse outcomes after intracerebral hemorrhage (ICH), a stroke subtype with no effective treatment. Despite the expected increase in the incidence of ICH due to population aging and the widespread use of anticoagulants, preclinical studies with aged animal subjects are lacking, and the pathophysiology of ICH in aged subjects has yet to be defined. Herein, we attempt to characterize the brain proteomic changes after ICH using an unbiased label- free quantitative proteomics approach and bioinformatics. To this end, aged male and female mice (18-24 months old) were subjected to sham/ICH. Mice were euthanized on day 3 post-surgery, and ipsilateral brain tissue was collected and subjected to LC-MS/MS analysis. Considering sex as a biological variable, the data derived from males and females were separately analyzed. The proteomics analysis revealed 133 differentially expressed proteins (DEPs) between the sham and ICH groups in male subjects. Among the DEPs, 98 proteins were downregulated, and 35 proteins were upregulated after ICH, compared to sham. In aged female mice, 315 DEPs were identified, of which 221 proteins were downregulated, and 94 proteins were upregulated after ICH compared to sham. The mass spectrometry data was validated using immunohistochemistry or western blot analysis, and the bioinformatics analysis revealed a comprehensive understanding of the signaling pathways associated with ICH. Some DEPs in both aged male and female mice that could play roles in ICH pathology were 14-3-3 proteins and S100-A9. The study also revealed that mitochondrial dysfunction could be a critical regulator of ICH-induced acute brain damage. Overall, the generated proteomics data could help develop hypothesis-driven functional analysis and delineate the complex pathobiology of ICH.
衰老脑出血（ICH）是一种没有有效治疗方法的中风亚型，是不良后果的独立预测因素。尽管由于人口老龄化和抗凝剂的广泛使用，预计ICH的发病率会增加，但缺乏对老年动物受试者的临床前研究，老年受试者ICH的病理生理学尚未明确。在此，我们试图使用无标记定量蛋白质组学方法和生物信息学来描述ICH后脑蛋白质组的变化。为此，对18-24个月大的老年雄性和雌性小鼠进行假手术/ICH手术。术后第3天对小鼠实施安乐死，收集同侧脑组织并进行液相色谱-串联质谱（LC-MS/MS）分析。将性别作为生物学变量，分别分析来自雄性和雌性的数据。蛋白质组学分析显示，雄性受试者假手术组和ICH组之间有133种差异表达蛋白（DEP）。在这些DEP中，与假手术组相比，ICH后98种蛋白下调，35种蛋白上调。在老年雌性小鼠中，鉴定出315种DEP，其中与假手术组相比，ICH后221种蛋白下调，94种蛋白上调。通过免疫组织化学或蛋白质印迹分析验证了质谱数据，生物信息学分析揭示了对与ICH相关信号通路的全面理解。在老年雄性和雌性小鼠中，一些可能在ICH病理学中起作用的DEP是14-3-3蛋白和S100-A9。该研究还表明，线粒体功能障碍可能是ICH诱导急性脑损伤的关键调节因子。总体而言，生成的蛋白质组学数据有助于开展基于假设的功能分析，并阐明ICH复杂的病理生物学机制。
