Abstract：
Menaquinone-4(MK-4), the isoform of vitamin K2 in the brain, exerts neuroprotective effects against a variety of central nervous system disorders. This study aimed to demonstrate the anti-ferroptosis effects of MK-4 in neurons after SAH.
A subarachnoid hemorrhage (SAH) model was prepared by endovascular perforation in mice. In vitro hemoglobin stimulation of primary cortical neurons mimicked SAH. MK-4, Brequinar (BQR, DHODH inhibitor), and Selisistat (SEL, SIRT1 inhibitor) were administered, respectively. Subsequently, WB, immunofluorescence was used to determine protein expression and localization, and transmission electron microscopy was used to observe neuronal mitochondrial structure while other indicators of ferroptosis were measured.
MK-4 treatment significantly upregulated the protein levels of DHODH; decreased GSH, PTGS2, NOX1, ROS, and restored mitochondrial membrane potential. Meanwhile, MK-4 upregulated the expression of SIRT1 and promoted its entry into the nucleus. BQR or SEL partially abolished the protective effect of MK-4 on, neurologic function, and ferroptosis.
Taken together, our results suggest that MK-4 attenuates ferroptosis after SAH by upregulating DHODH through the activation of SIRT1.
A subarachnoid hemorrhage (SAH) model was prepared by endovascular perforation in mice. In vitro hemoglobin stimulation of primary cortical neurons mimicked SAH. MK-4, Brequinar (BQR, DHODH inhibitor), and Selisistat (SEL, SIRT1 inhibitor) were administered, respectively. Subsequently, WB, immunofluorescence was used to determine protein expression and localization, and transmission electron microscopy was used to observe neuronal mitochondrial structure while other indicators of ferroptosis were measured.
MK-4 treatment significantly upregulated the protein levels of DHODH; decreased GSH, PTGS2, NOX1, ROS, and restored mitochondrial membrane potential. Meanwhile, MK-4 upregulated the expression of SIRT1 and promoted its entry into the nucleus. BQR or SEL partially abolished the protective effect of MK-4 on, neurologic function, and ferroptosis.
Taken together, our results suggest that MK-4 attenuates ferroptosis after SAH by upregulating DHODH through the activation of SIRT1.
摘要：
背景：Menaquinone-4（MK-4），大脑中维生素K2的同工型，对多种中枢神经系统疾病发挥神经保护作用。本研究旨在证明MK-4在SAH后神经元中的抗铁凋亡作用。
方法：通过血管内穿孔制备小鼠蛛网膜下腔出血（SAH）模型。体外血红蛋白刺激初级皮质神经元模拟SAH。MK-4，布基那（BQR，DHODH抑制剂)，和Selissistat(SEL，SIRT1抑制剂）被施用，分别。随后，WB,免疫荧光用于确定蛋白质表达和定位，用透射电镜观察神经元线粒体结构，同时测量铁性凋亡的其他指标。
结果：MK-4处理显著上调DHODH蛋白水平；GSH降低，PTGS2,NOX1,ROS,并恢复了线粒体膜电位.同时,MK-4上调SIRT1的表达并促进其进入细胞核。BQR或SEL部分取消了MK-4对，神经功能,和铁中毒。
结论：综合来看，我们的结果表明，MK-4通过激活SIRT1上调DHODH，从而减弱SAH后的铁凋亡。
方法：通过血管内穿孔制备小鼠蛛网膜下腔出血（SAH）模型。体外血红蛋白刺激初级皮质神经元模拟SAH。MK-4，布基那（BQR，DHODH抑制剂)，和Selissistat(SEL，SIRT1抑制剂）被施用，分别。随后，WB,免疫荧光用于确定蛋白质表达和定位，用透射电镜观察神经元线粒体结构，同时测量铁性凋亡的其他指标。
结果：MK-4处理显著上调DHODH蛋白水平；GSH降低，PTGS2,NOX1,ROS,并恢复了线粒体膜电位.同时,MK-4上调SIRT1的表达并促进其进入细胞核。BQR或SEL部分取消了MK-4对，神经功能,和铁中毒。
结论：综合来看，我们的结果表明，MK-4通过激活SIRT1上调DHODH，从而减弱SAH后的铁凋亡。
