Cai Jun, Liu Yuxuan, Gong Li, Deng Shengfeng, Zheng Wenhua, Li Shuai
Innovation Center of Cardiometabolic Diseases, Guangdong Medical University, Dongguan, China.
Guangdong Provincial Engineering Research Center of Molecular Imaging, The Fifth Affiliated Hospital of Sun Yat-Sen University, Zhuhai, China.
Mol Neurobiol. 2025 Jun 10. doi: 10.1007/s12035-025-05141-8.
Alzheimer's disease (AD) represents the most common manifestation of dementia. Cerebrovascular dysfunction constitutes one of the initial events in the pathogenic process of AD. Recently, our research group has reported that artemether, a first-line antimalarial drug approved by the FDA, exhibits a neuroprotective property. Nevertheless, the impact of artemether on cerebrovascular dysfunction, along with its underlying mechanisms, still awaits comprehensive elucidation. This study systematically investigated the neuroprotective effects of artemether against Aβ-induced injury in brain microvascular endothelial cells and explored the underlying molecular mechanisms. Our findings demonstrate that artemether potently mitigates Aβ-mediated cytotoxicity and endothelial barrier dysfunction in mouse brain microvascular endothelial cells. Moreover, artemether attenuated blood-brain barrier disruption by upregulating tight junction proteins OCLN, CLDN-5, and ZO-1. Further studies revealed that artemether treatment improved elevated ROS levels, the dissipation of mitochondrial membrane potential, and the decrease of ATP content. Immunoblotting analysis demonstrated that artemether stimulated the CAMKK2-AMPK kinase and its downstream target PGC1α. In addition, the blockage of the CAMKK2/AMPK/PGC1α pathway by inhibitor STO-609, Compound C, or knocking down the expression level of PGC1α with sgRNA significantly attenuated the neuroprotective activity of artemether. These results indicate the essential role of the CAMKK2/AMPK/PGC1α pathway in the protective effect of artemether in mouse brain microvascular endothelial cells. Similar results were obtained in Aβ-treated human brain microvascular endothelial cells. In the AD mice model, we found that artemether treatment increased the expression level of ZO-1, CLDN-5, and OCLN of the AD mice model. Collectively, these findings demonstrate that artemether exerts neuroprotection against Aβ-induced injury in brain microvascular endothelial cells through activation of the CAMKK2/AMPK/PGC1α signaling axis, which supports the protective effect of artemether in the prevention and treatment of AD through improving cerebrovascular dysfunction.
阿尔茨海默病（AD）是痴呆最常见的表现形式。脑血管功能障碍是AD致病过程中的初始事件之一。最近，我们的研究小组报道，美国食品药品监督管理局（FDA）批准的一线抗疟药物蒿甲醚具有神经保护特性。然而，蒿甲醚对脑血管功能障碍的影响及其潜在机制仍有待全面阐明。本研究系统地研究了蒿甲醚对Aβ诱导的脑微血管内皮细胞损伤的神经保护作用，并探讨了其潜在的分子机制。我们的研究结果表明，蒿甲醚能有效减轻小鼠脑微血管内皮细胞中Aβ介导的细胞毒性和内皮屏障功能障碍。此外，蒿甲醚通过上调紧密连接蛋白OCLN、CLDN - 5和ZO - 1减轻血脑屏障破坏。进一步研究表明，蒿甲醚治疗改善了升高的ROS水平、线粒体膜电位的耗散和ATP含量的降低。免疫印迹分析表明，蒿甲醚刺激了CAMKK2 - AMPK激酶及其下游靶点PGC1α。此外，用抑制剂STO - 609、Compound C阻断CAMKK2 / AMPK / PGC1α通路，或用sgRNA敲低PGC1α的表达水平，均显著减弱了蒿甲醚的神经保护活性。这些结果表明CAMKK2 / AMPK / PGC1α通路在蒿甲醚对小鼠脑微血管内皮细胞的保护作用中起重要作用。在Aβ处理的人脑微血管内皮细胞中也获得了类似的结果。在AD小鼠模型中，我们发现蒿甲醚治疗增加了AD小鼠模型中ZO - 家族性高胆固醇血症（FH）是一种常染色体显性疾病，由低密度脂蛋白受体（LDLR）、载脂蛋白B（APOB）、前蛋白转化酶枯草溶菌素9（PCSK9）或低密度脂蛋白受体衔接蛋白1（LDLRAP1）等基因的功能丧失突变引起。FH的特征是低密度脂蛋白胆固醇（LDL-C）水平极高，早发性心血管疾病风险增加。目前的指南建议对FH患者进行高强度他汀类药物治疗，目标是将LDL-C降低至少50%。然而，许多患者未能达到这一目标，需要额外的降脂治疗。单克隆抗体已成为FH治疗的重要组成部分，特别是那些对他汀类药物不耐受或反应不足的患者。在这项研究中，我们评估了一种新型单克隆抗体evolocumab在杂合子FH患者中的疗效和安全性。
我们进行了一项多中心、双盲、安慰剂对照试验，纳入了141例杂合子FH患者。患者被随机分配接受evolocumab（420 mg皮下注射，每2周一次）或安慰剂治疗24周。主要终点是第24周时LDL-C从基线的百分比变化。次要终点包括其他血脂参数的变化、安全性和耐受性。
结果显示，与安慰剂组相比，evolocumab组的LDL-C水平显著降低（平均降低59.5% vs 安慰剂组的平均降低0.9%）。evolocumab组还观察到其他血脂参数的显著改善，包括总胆固醇、甘油三酯和载脂蛋白B水平的降低，以及高密度脂蛋白胆固醇水平的升高。在安全性方面，evolocumab组和安慰剂组之间没有观察到显著差异。最常见的不良事件是注射部位反应，两组的发生率相似。
这些结果表明，evolocumab在杂合子FH患者中是一种有效且安全的降脂治疗方法。它显著降低了LDL-C水平，并改善了其他血脂参数，且安全性良好。这些发现为FH的治疗提供了一种新的选择，特别是对于那些对他汀类药物治疗反应不足的患者。 1、CLDN - 5和OCLN的表达水平。总体而言，这些发现表明，蒿甲醚通过激活CAMKK2 / AMPK / PGC1α信号轴对Aβ诱导的脑微血管内皮细胞损伤发挥神经保护作用，这支持了蒿甲醚通过改善脑血管功能障碍在AD预防和治疗中的保护作用。
