依泽替米贝，一种 NPC1L1 抑制剂，通过 MCAO 后 AMPK 依赖性自噬激活减轻大鼠神经元凋亡。
Ezetimibe, a NPC1L1 inhibitor, attenuates neuronal apoptosis through AMPK dependent autophagy activation after MCAO in rats.
机构信息
Department of Anesthesiology, The Second Affiliated Hospital of Zhejiang University, School of Medicine, Hangzhou, Zhejiang, China; Department of Anesthesiology and Basic Sciences, Loma Linda University, School of Medicine, Loma Linda, CA, USA.
Department of Anesthesiology and Basic Sciences, Loma Linda University, School of Medicine, Loma Linda, CA, USA.
出版信息
Exp Neurol. 2018 Sep;307:12-23. doi: 10.1016/j.expneurol.2018.05.022. Epub 2018 May 28.
Autophagy activation exerts neuroprotective effects in the ischemic stroke model. Ezetimibe (Eze), a Niemann-Pick disease type C1-Like 1 (NPC1L1) pharmacological inhibitor, has been reported to protect hepatocytes from apoptosis via autophagy activation. In this study, we explored whether Eze could attenuate neuronal apoptosis in the rat model of middle cerebral artery occlusion (MCAO), specifically via activation of the AMPK/ULK1/autophagy pathway. Two hundred and one male Sprague-Dawley rats were subjected to transient MCAO followed by reperfusion. Eze was administered 1 h after MCAO. To elucidate the underlying molecular mechanism, Dorsomorphin, a selective AMPK inhibitor, and 3-methyladenine (3-MA), an autophagy inhibitor, were injected intracerebroventricularly before MCAO. Infarct volume, neurological score, brain cholesterol levels, immunofluorescence staining, Western blot, and Fluoro-Jade C (FJC) staining were used to evaluate the effects of Eze. The endogenous NPC1L1 expression increased and mainly expressed in neurons after MCAO. Intranasal administration of Eze reduced brain infarct volume at 24 and 72 h after MCAO, with improved short and long-term neurological functions after MCAO. Eze reduced brain cholesterol levels (total cholesterol, free cholesterol and cholesteryl esters) and the number of FJC-positive neurons. The expression of phosphorylated AMPK (p-AMPK) and downstream ULK1, Beclin1, LC3BII, Bcl-2, and Bcl-xl increased, while P62 and proapoptotic Bax decreased after treatment with Eze. Pretreatment with Dorsomorphin and 3-MA reversed the beneficial effects of Eze. These findings suggest that intranasal administration of Eze plays neuroprotective role through autophagy activation after MCAO in rats. Lowered cholesterol levels and AMPK activation may act in conjunction to induce autophagy after treatment with Eze. Eze merits further investigation as a potential therapeutic agent in ischemic stroke patients.
自噬激活在缺血性中风模型中发挥神经保护作用。依泽替米贝（Eze）是一种尼曼-匹克病 C1 样 1（NPC1L1）药理学抑制剂，据报道它通过自噬激活来保护肝细胞免于凋亡。在本研究中，我们探讨了 Eze 是否可以通过激活 AMPK/ULK1/自噬通路来减轻大鼠大脑中动脉闭塞（MCAO）模型中的神经元凋亡。201 只雄性 Sprague-Dawley 大鼠接受短暂性 MCAO 后继发再灌注。MCAO 后 1 小时给予 Eze。为了阐明潜在的分子机制，在 MCAO 前通过脑室内注射 Dorsomorphin（一种选择性 AMPK 抑制剂）和 3-甲基腺嘌呤（3-MA，一种自噬抑制剂）。使用脑梗死体积、神经功能评分、脑内胆固醇水平、免疫荧光染色、Western blot 和 Fluoro-Jade C（FJC）染色来评估 Eze 的作用。MCAO 后，内源性 NPC1L1 表达增加，主要在神经元中表达。鼻内给予 Eze 可减少 MCAO 后 24 和 72 小时的脑梗死体积，并改善 MCAO 后的短期和长期神经功能。Eze 降低了脑内胆固醇水平（总胆固醇、游离胆固醇和胆固醇酯）和 FJC 阳性神经元的数量。用 Eze 处理后，磷酸化 AMPK（p-AMPK）及其下游的 ULK1、Beclin1、LC3BII、Bcl-2 和 Bcl-xl 的表达增加，而 P62 和促凋亡 Bax 减少。用 Dorsomorphin 和 3-MA 预处理可逆转 Eze 的有益作用。这些发现表明，在大鼠 MCAO 后，鼻内给予 Eze 通过自噬激活发挥神经保护作用。胆固醇水平降低和 AMPK 激活可能共同作用于 Eze 治疗后诱导自噬。Eze 作为缺血性中风患者的潜在治疗药物值得进一步研究。
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