Yunnan Key Laboratory of Dai and Yi Medicine, Yunnan University of Chinese Medicine, Kunming, Yunnan 650500, P.R. China.
Mol Med Rep. 2023 Jan;27(1). doi: 10.3892/mmr.2022.12896. Epub 2022 Nov 11.
3,4‑Dihydroxybenzaldehyde (DBD), one of the active components of has a cerebral protective effect and can effectively combat cerebral ischemia/reperfusion (I/R) injury in rats. However, the metabolite profiles and underlying mechanisms associated with DBD remain unclear. To explore the level of energy metabolism and pharmacological targets in brain tissue following DBD treatment of stroke. The right middle cerebral artery of the rats was occluded for 2 h and reperfused for 24 h to simulate brain I/R injury. Pharmacological results showed that DBD reduced cerebral infarct volume, improved neurological function and increased adenosine triphosphate (ATP) content. Mitochondria are the primary sites for ATP generation and cellular energy supply and decreasing mitochondrial dysfunction can alleviate the energy expenditure of ischemic stroke. Through further experiments, it was found that mitochondrial damage was recovered following DBD treatment, which was manifested by the improvement of mitochondrial morphology under an electron microscope and the reduction of oxidative stress damage. The metabolomics of middle cerebral artery occlusion/reperfusion (MCAO/R) rat brain tissue was studied by the liquid chromatography‑tandem mass spectrometry metabolomics method. Significantly different metabolites were screened and the pathways involved included amino sugar and nucleotide sugar metabolism and pentose phosphate pathway. Finally, the present study performed targeted metabolic profiling and validated potential therapeutic targets. Uridine diphosphate N‑acetylglucosamine (UDP‑GlcNAc) levels were decreased in the MCAO/R group but significantly increased in the DBD group. TdT‑mediated dUTP nick end labeling (TUNEL) staining, hematoxylin and eosin staining and western blotting showed that brain cell apoptosis was inhibited and neuronal morphology improved. Among them, the regulatory enzyme O‑GlcNAc transferase (OGT) of UDP‑GlcNAc appeared to be significantly increased and neuronal apoptosis was inhibited following DBD treatment, which was verified by western blotting. Therefore, by analyzing mitochondrial dysfunction following I/R and the characterization of potential markers in mitochondrial energy metabolism, it was shown that OGT could inhibit neuronal apoptosis as a potential therapeutic target for brain I/R injury.
3,4-二羟基苯甲醛（DBD）是 的一种活性成分，具有脑保护作用，可有效防治大鼠脑缺血/再灌注（I/R）损伤。然而，DBD 相关的代谢物谱和潜在机制尚不清楚。本研究旨在探讨 DBD 治疗中风后脑组织能量代谢和药理学靶点的变化。通过阻断大鼠右侧大脑中动脉 2 h 再灌注 24 h 来模拟脑 I/R 损伤。药理结果表明，DBD 可减少脑梗死体积，改善神经功能，增加三磷酸腺苷（ATP）含量。线粒体是产生 ATP 和细胞能量供应的主要场所，减轻线粒体功能障碍可减轻缺血性中风的能量消耗。进一步实验发现，DBD 治疗后线粒体损伤得到恢复，电镜下观察到线粒体形态改善，氧化应激损伤减轻。采用液相色谱-串联质谱代谢组学方法研究大脑中动脉闭塞/再灌注（MCAO/R）大鼠脑组织的代谢组学。筛选出差异有统计学意义的代谢物，涉及的代谢途径包括氨基糖和核苷酸糖代谢以及戊糖磷酸途径。最后，本研究进行了靶向代谢组学分析并验证了潜在的治疗靶点。MCAO/R 组尿苷二磷酸-N-乙酰氨基葡萄糖（UDP-GlcNAc）水平降低，而 DBD 组显著升高。TdT 介导的 dUTP 缺口末端标记（TUNEL）染色、苏木精-伊红（H&E）染色和 Western blot 显示，DBD 可抑制脑细胞凋亡，改善神经元形态。其中，UDP-GlcNAc 的调节酶 O-连接的 N-乙酰氨基葡萄糖转移酶（OGT）在 DBD 治疗后明显增加，神经元凋亡受到抑制，Western blot 也得到了验证。因此，通过分析 I/R 后线粒体功能障碍和线粒体能量代谢潜在标志物的特征，表明 OGT 可作为脑 I/R 损伤的潜在治疗靶点抑制神经元凋亡。
