PCMT1 Ameliorates Neuronal Apoptosis by Inhibiting the Activation of MST1 after Subarachnoid Hemorrhage in Rats
PCMT1 Ameliorates Neuronal Apoptosis by Inhibiting the Activation of MST1 after Subarachnoid Hemorrhage in Rats
复制标题
PCMT1 通过抑制大鼠蛛网膜下腔出血后 MST1 的激活来改善神经元凋亡
DOI：
10.1007/s12975-017-0540-8
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发表时间：
2017-10-01
影响因子：
6.9
通讯作者：
Zhang, Jianmin
中科院分区：
文献类型：
--
作者：
Shi, Ligen;Al-Baadani, Ammar;Zhang, Jianmin
哺乳动物不育20-样蛋白1(MST1)被发现促进神经细胞的凋亡。蛋白-L-异门冬氨酸(D-天冬氨酸)O-甲基转移酶(PCMT1)是最近发现的一种与MST1相互作用的蛋白，是一种抗细胞凋亡因子。本研究旨在探讨PCMT1在减少大鼠蛛网膜下腔出血(SAH)后MST1诱导的神经细胞凋亡中的作用。实验选用雄性SD大鼠198只。血管内穿孔SAH后1h皮下注射外源性PCMT1激动剂CGP 3466B。SAH后即刻经侧脑室注射白屈菜红碱或盏花素A。分别于SAH后24和72h测定SAH分级、Garcia评分和脑含水量。免疫荧光法检测神经细胞凋亡。免疫印迹法检测内源性PCMT1、MST1、磷酸化MST1(p-MST1)、裂解MST1(CL-MST1)及细胞凋亡相关蛋白的表达水平。SAH后PCMT1、MST1表达减少，caspase3活性升高。CGP 3466B治疗可改善神经行为功能，减少脑含水量，抑制MST1活性，减轻神经细胞凋亡。这些神经保护作用可通过促进Calyculin A的MST1磷酸化或白屈菜红碱增加CL-MST1而显著减弱。PCMT1通过降低MST1的磷酸化和CL-MST1的水平来抑制神经细胞的凋亡。PCMT1/MST1通路可能是减轻SAH后早期脑损伤的替代治疗靶点。
Mammalian sterile 20-like kinase 1 (MST1) is found to promote neuronal apoptosis. Protein-l-isoaspartate (d-aspartate)O-methyltransferase (PCMT1), an anti-apoptosis factor, was recently identified as an MST1-interacting protein. This study aims to explore the potential role of PCMT1 in reducing MST1-induced neuronal apoptosis after subarachnoid hemorrhage (SAH) in rats. One hundred ninety-eight male Sprague-Dawley rats were used. An exogenous PCMT1 agonist, CGP 3466B, was injected subcutaneously 1 h after the SAH induced by endovascular perforation. Chelerythrine or calyculin A was given immediately via intracerebroventricular administration after SAH. The SAH grade, Garcia score, and brain water content were measured at 24 and 72 h after the SAH. Neuronal apoptosis was detected by an immunofluorescent assay. The expression levels of endogenous PCMT1, MST1, phospho-MST1 (p-MST1), cleaved MST1 (cl-MST1), and apoptosis-related proteins were studied by western blotting. The expression of PCMT1 and MST1 decreased, while the level of active caspase 3 increased in rats after SAH. CGP 3466B treatment improved neurobehavioral function, reduced brain water content, inhibited the activity of MST1, and relieved neuronal apoptosis. These neuroprotective effects were significantly weakened either through accelerating MST1 phosphorylation by calyculin A or increasing cl-MST1 by chelerythrine. PCMT1 inhibited neuronal apoptosis by reducing MST1 phosphorylation and the level of cl-MST1. PCMT1/MST1 pathway might be an alternative therapeutic target for alleviating early brain injury after SAH.