College of Pharmacology, Fujian University of Traditional Chinese Medicine, No.1, Qiu Yang Road, Min Hou Shang Jie, Fuzhou, 350122, China.
Dept of Urology, the First Affiliated Hospital of Fujian Medical University, Fuzhou 350005, China; Dept of Urology, National Region Medical Center, Binhai Campus of the First Affiliated Hospital, Fujian Medical University, Fuzhou 350212,China; Fujian Institute of Urology, the First Affiliated Hospital,Fujian Medical University, Fuzhou 350005, China.
Phytomedicine. 2024 Dec;135:156208. doi: 10.1016/j.phymed.2024.156208. Epub 2024 Nov 9.
Ischemic stroke is a common cerebrovascular disease characterized by high incidence, disability, mortality, and recurrence. The limitations of current pharmacological treatments, which have primarily single neuroprotective action and a narrow therapeutic time window, lead to unsatisfactory therapeutic efficacy. Activation of autophagy can facilitate neural regeneration.
To clarify whether salidroside can promote axonal sprouting through autophagy resulting in protecting neurons.
In vivo, a Middle Cerebral Artery Occlusion/reperfusion (MCAO/IR) model was used, and in vitro, an Oxygen-Glucose Deprivation/Reoxygenation (OGD/R)-induced primary neuronal cell model was employed to evaluate the neuroprotective effects of salidroside. BDA neurotracer, immunofluorescence, and Western blot (WB) were utilized to determine its impact on axonal sprouting and the levels of related proteins (MAP2, GAP43, and PSD-95). Proteomics, transmission electron microscopy (TEM), and WB were applied to identify the effects on autophagy-related proteins (beclin1, LC3, p62, and LAMP2), autophagosomes and lysosomes. The mechanism of salidroside in promoting axonal sprouting through inducing autophagy was further confirmed by blocking with the autophagy inhibitor 3-MA.
Salidroside reduced neurologic deficits and infarct volume induced by MCAO/IR in vivo and protected OGD/R induced primary neuronal cells in vitro. Both in vivo and in vitro, it increased the number and length of axons and upregulated the expression of key axonal proteins (MAP2, GAP43, and PSD-95) and mediated autophagy-related proteins. Mechanistic studies showed that the promoting effects of salidroside on autophagy and axonal sprouting disappeared after the blockade by 3-MA.
This study reports for the first time that the neuroprotective effect of salidroside in ischemic stroke can be executed through mediating autophagy-related protein (beclin1, LC3, p62, and LAMP2), resulting in induced axonal sprouting or mature protein (MAP2, GAP43, and PSD-95).
缺血性脑卒中是一种常见的脑血管疾病，具有发病率高、致残率高、死亡率高和复发率高的特点。目前的药物治疗方法主要具有单一的神经保护作用，且治疗时间窗较窄，导致治疗效果不理想。自噬的激活可以促进神经再生。
阐明红景天苷是否可以通过自噬促进轴突发芽，从而保护神经元。
在体内，采用大脑中动脉闭塞/再灌注（MCAO/IR）模型，在体外，采用氧-葡萄糖剥夺/再氧合（OGD/R）诱导的原代神经元细胞模型来评估红景天苷的神经保护作用。使用 BDA 神经示踪剂、免疫荧光和 Western blot（WB）来确定其对轴突发芽和相关蛋白（MAP2、GAP43 和 PSD-95）水平的影响。蛋白质组学、透射电子显微镜（TEM）和 WB 用于鉴定自噬相关蛋白（beclin1、LC3、p62 和 LAMP2）、自噬体和溶酶体的影响。通过使用自噬抑制剂 3-MA 阻断，进一步证实了红景天苷通过诱导自噬促进轴突发芽的机制。
红景天苷减少了体内 MCAO/IR 引起的神经功能缺损和梗死体积，以及体外 OGD/R 诱导的原代神经元细胞损伤。在体内和体外，它都增加了轴突的数量和长度，并上调了关键轴突蛋白（MAP2、GAP43 和 PSD-95）和介导自噬相关蛋白的表达。机制研究表明，在 3-MA 阻断后，红景天苷对自噬和轴突发芽的促进作用消失。
本研究首次报道，红景天苷在缺血性脑卒中的神经保护作用可以通过调节自噬相关蛋白（beclin1、LC3、p62 和 LAMP2）来实现，从而诱导轴突发芽或成熟蛋白（MAP2、GAP43 和 PSD-95）。
