Department of Neurobiology, School of Basic Medicine, Jinzhou Medical University, Jinzhou, Liaoning, China.
Institute of Life Science, Jinzhou Medical University, Jinzhou, Liaoning, China.
CNS Neurosci Ther. 2024 Sep;30(9):e70030. doi: 10.1111/cns.70030.
Ischemic stroke leads a primary cause of mortality in human diseases, with a high disability rate worldwide. This study aims to investigate the function of β-1,4-galactosyltransferase 1 (B4galt1) in mouse brain ischemia/reperfusion (I/R) injury.
Recombinant human B4galt1 (rh-B4galt1) was intranasally administered to the mice model of middle cerebral artery occlusion (MCAO)/reperfusion. In this study, the impact of rh-B4galt1 on cerebral injury assessed using multiple methods, including the neurological disability status scale, 2,3,5-triphenyltetrazolium chloride (TTC), Nissl and TUNEL staining. This study utilized laser speckle Doppler flowmeter to monitor the cerebral blood flow. Western blotting was performed to assess the protein expression levels, and fluorescence-labeled dihydroethidium method was performed to determine the superoxide anion generation. Assay kits were used for the measurement of iron, malondialdehyde (MDA) and glutathione (GSH) levels.
We demonstrated that rh-B4galt1 markedly improved neurological function, reduced cerebral infarct volume and preserved the completeness of blood-brain barrier (BBB) for preventing damage. These findings further illustrated that rh-B4galt1 alleviated oxidative stress, lipid peroxidation, as well as iron deposition induced by I/R. The vital role of ferroptosis was proved in brain injury. Furthermore, the rh-B4galt1 could increase the levels of TAZ, Nrf2 and HO-1 after I/R. And TAZ-siRNA and ML385 reversed the neuroprotective effects of rh-B4galt1.
The results indicated that rh-B4galt1 implements neuroprotective effects by modulating ferroptosis, primarily via upregulating TAZ/Nrf2/HO-1 pathway. Thus, B4galt1 could be seen as a promising novel objective for ischemic stroke therapy.
缺血性中风是人类疾病中主要的致死原因，在全球范围内具有较高的致残率。本研究旨在探讨β-1,4-半乳糖基转移酶 1（B4galt1）在小鼠脑缺血/再灌注（I/R）损伤中的作用。
重组人 B4galt1（rh-B4galt1）经鼻腔给予大脑中动脉闭塞（MCAO）/再灌注小鼠模型。在这项研究中，采用神经功能缺损评分、2,3,5-三苯基氯化四氮唑（TTC）、尼氏染色和 TUNEL 染色等多种方法评估 rh-B4galt1 对脑损伤的影响。采用激光散斑多普勒血流仪监测脑血流，Western blot 检测蛋白表达水平，荧光标记二氢乙啶法检测超氧阴离子生成，试剂盒检测铁、丙二醛（MDA）和谷胱甘肽（GSH）水平。
我们发现 rh-B4galt1 可显著改善神经功能，减少脑梗死体积，维持血脑屏障（BBB）的完整性，从而防止损伤。这些发现进一步表明，rh-B4galt1 减轻了 I/R 引起的氧化应激、脂质过氧化和铁沉积。铁死亡在脑损伤中起着重要作用。此外，rh-B4galt1 可增加 I/R 后 TAZ、Nrf2 和 HO-1 的水平。TAZ-siRNA 和 ML385 逆转了 rh-B4galt1 的神经保护作用。
结果表明，rh-B4galt1 通过调节铁死亡发挥神经保护作用，主要通过上调 TAZ/Nrf2/HO-1 通路。因此，B4galt1 可作为缺血性中风治疗的一个有前途的新靶点。
