Wu Yuanhua, Ren Yanna, Cao Liping, Zhao Jianghu, Cai Jing, Wang Azhen, Tian Maoping, Li Yueli, Zheng Yijia, Shi Jingyu, Lei Tong
Department of neurology, The First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine, Guiyang 550002, Guizhou, China.
Department of Psychology, College of Humanities and Management, Guizhou University of Traditional Chinese Medicine, Guiyang 550025, Guizhou, China.
Int Immunopharmacol. 2025 Aug 22;164:115398. doi: 10.1016/j.intimp.2025.115398.
As a common cognitive impairment disease, vascular dementia (VD) belongs to the category of "dementia" in Chinese medicine, and should be treated with the principle "Governor Vessel" as the primary focus. The occurrence of VD is closely related to cerebral ischemia, hippocampal neuron damage, and excessive mitochondrial autophagy activation, which has been considered as is a key mechanism of PINK1/Parkin signaling pathway regulation. Our study aims to investigate molecular mechanisms of the Governor Vessel acupuncture method in improving VD cognitive dysfunction by regulating mitochondrial autophagy.
The study utilized 310 healthy adult male Sprague-Dawley (SD) rats (SPF-grade, aged 18 months, weighing 500-700 g). Vascular dementia (VD) model was established using permanent bilateral common carotid artery ligation (2VO). Additionally, adenoviral vector intervention and drug intervention groups were set up to explore the mechanistic pathways. Morris water maze and shuttle box tests were used to assess cognitive function. Hippocampal tissue changes were analyzed through Nissl staining, HE staining, TUNEL (cell apoptosis), JC-1 (mitochondrial membrane potential), qPCR, and Western blot (to detect PINK1/Parkin pathway proteins). Data are presented as mean ± SEM, analyzed by one-way ANOVA (for normal distribution) or Friedman test (for non-normal distribution), with Tukey's post-hoc test. p < 0.05 was considered statistically significant.
Rats in the VD model group showed significant cognitive decline in behavioral evaluations, while neurons in the hippocampal CA1 region exhibited varying degrees of damage and apoptosis. Besides, the expressions of autophagy-related proteins, Beclin-1, p-Parkin/Parkin, PINK1, and LC3 II/I was notably up-regulated, whereas the expression of p62 and Mfn2 proteins was significantly decreased. After governor vessel acupuncture, the cognitive ability of VD rats was improved significantly, and neuronal damage in hippocampal CA1 region was effectively alleviated, indicating that governor vessel acupuncture inhibited autophagy by down-regulating the PINK1/Parkin signaling pathway, thereby alleviating VD. Further, PINK1 viral vector and drug intervention experiments further confirmed that inhibiting autophagy could effectively protect neurons by down-regulating the PINK1/Parkin signaling pathway.
Governor vessel acupuncture can effectively protect hippocampal neurons and enhance cognitive functions in VD models by down-regulating PINK1/Parkin signaling pathway to inhibit mitochondrial autophagy, providing a theoretical basis for its clinical application.
血管性痴呆（VD）作为一种常见的认知障碍疾病，在中医范畴中属于“痴呆”，应以“督脉”理论为主要依据进行治疗。VD的发生与脑缺血、海马神经元损伤以及过度激活的线粒体自噬密切相关，这被认为是PINK1/Parkin信号通路调控的关键机制。本研究旨在探讨督脉针刺法通过调节线粒体自噬改善VD认知功能障碍的分子机制。
本研究选用310只健康成年雄性Sprague-Dawley（SD）大鼠（SPF级，18月龄，体重500 - 700 g）。采用永久性双侧颈总动脉结扎（2VO）建立血管性痴呆（VD）模型。此外，设置腺病毒载体干预组和药物干预组以探究作用机制途径。通过Morris水迷宫和穿梭箱试验评估认知功能。通过尼氏染色、HE染色、TUNEL（细胞凋亡）、JC-1（线粒体膜电位）、qPCR和蛋白质免疫印迹法（检测PINK1/Parkin通路蛋白）分析海马组织变化。数据以平均值±标准误表示，采用单因素方差分析（用于正态分布）或Friedman检验（用于非正态分布）进行分析，并进行Tukey事后检验。p < 0.05被认为具有统计学意义。
VD模型组大鼠在行为学评估中表现出明显的认知功能下降，海马CA1区神经元出现不同程度的损伤和凋亡。此外，自噬相关蛋白Beclin-1、p-Parkin/Parkin、PINK1和LC3 II/I的表达显著上调，而p62和Mfn2蛋白的表达显著降低。督脉针刺后，VD大鼠的认知能力显著提高，海马CA1区神经元损伤得到有效缓解，表明督脉针刺通过下调PINK1/Parkin信号通路抑制自噬，从而减轻VD。此外，PINK1病毒载体和药物干预实验进一步证实，抑制自噬可通过下调PINK1/Parkin信号通路有效保护神经元。
督脉针刺可通过下调PINK1/Parkin信号通路抑制线粒体自噬，有效保护VD模型中的海马神经元并增强认知功能，为其临床应用提供理论依据。
