Wang Panqin, Liu Yongyan, Du Yimeng, Gao Yiwen, Shao Tian, Guo Weifeng, Wang Zhenlong, Cheng Han
School of Life Sciences, Zhengzhou University, Zhengzhou 450001, China.
School of Electrical and Information Engineering, Zhengzhou University, Zhengzhou 450001, China.
Cells. 2025 Apr 1;14(7):527. doi: 10.3390/cells14070527.
Rapid ascent to high altitudes by unacclimatized individuals significantly increases the risk of brain damage, given the brain's heightened sensitivity to hypoxic conditions. Investigating hypoxia-tolerant animals can provide insights into adaptive mechanisms and guide prevention and treatment of hypoxic-ischemic brain injury. In this study, we exposed Brandt's voles to simulated altitudes (100 m, 3000 m, 5000 m, and 7000 m) for 24 h and performed quantitative proteomic and phosphoproteomic analyses of brain tissue. A total of 3990 proteins and 9125 phosphorylation sites (phospho-sites) were quantified. Differentially expressed (DE) analysis revealed that while protein abundance changes were relatively modest, phosphorylation levels exhibited substantial alterations, suggesting that Brandt's voles rapidly regulate protein structure and function through phosphorylation to maintain cellular homeostasis under acute hypoxia. Clustering analysis showed that most co-expressed proteins exhibited non-monotonic responses with increasing altitude, which were enriched in pathways related to cytokine secretion regulation and glutathione metabolism, contributing to reduced inflammation and oxidative stress. In contrast, most co-expressed phospho-sites showed monotonic changes, with phospho-proteins enriched in glycolysis and vascular smooth muscle contraction regulation. Kinase activity prediction identified nine hypoxia-responsive kinases, four of which belonging to the CAMK family. Immunoblot validated that the changes in CAMK2A activity were consistent with predictions, suggesting that CAMK may play a crucial role in hypoxic response. In conclusion, this work discovered that Brandt's voles may cope with hypoxia through three key strategies: (1) vascular regulation to enhance cerebral blood flow, (2) glycolytic activation to increase energy production, and (3) activation of neuroprotective mechanisms.
由于大脑对缺氧状况的敏感性增强，未适应环境的个体快速上升到高海拔地区会显著增加脑损伤风险。研究耐缺氧动物有助于深入了解适应机制，并为缺氧缺血性脑损伤的预防和治疗提供指导。在本研究中，我们将布氏田鼠暴露于模拟海拔高度（100米、3000米、5000米和7000米）24小时，并对脑组织进行定量蛋白质组学和磷酸化蛋白质组学分析。共定量了3990种蛋白质和9125个磷酸化位点。差异表达分析显示，虽然蛋白质丰度变化相对较小，但磷酸化水平却有显著改变，这表明布氏田鼠在急性缺氧条件下通过磷酸化快速调节蛋白质结构和功能以维持细胞内稳态。聚类分析表明，大多数共表达蛋白质随海拔升高呈现非单调反应，这些蛋白质富集于与细胞因子分泌调节和谷胱甘肽代谢相关的通路，有助于减轻炎症和氧化应激。相比之下，大多数共表达磷酸化位点呈现单调变化，磷酸化蛋白质富集于糖酵解和血管平滑肌收缩调节通路。激酶活性预测确定了9种缺氧反应激酶，其中4种属于钙/钙调蛋白依赖性蛋白激酶（CAMK）家族。免疫印迹验证了CAMK2A活性的变化与预测一致，表明CAMK可能在缺氧反应中起关键作用。总之，本研究发现布氏田鼠可能通过三种关键策略应对缺氧：（1）血管调节以增加脑血流量；（2）激活糖酵解以增加能量产生；（3）激活神经保护机制。
