Yang Miao, Yu Wen-Jing, He Chun-Xiang, Jin Yi-Jie, Li Ze, Li Ping, Deng Si-Si, Yi Ya-Qiao, Cheng Shao-Wu, Song Zhen-Yan
Hunan Province Key Laboratory of Cerebrovascular Disease Prevention and Treatment of Integrated Traditional Chinese and Western Medicine, Hunan University of Chinese Medicine Changsha 410208, China.
Zhongguo Zhong Yao Za Zhi. 2023 Jan;48(2):534-541. doi: 10.19540/j.cnki.cjcmm.20220907.501.
This study investigated the mechanism of Danggui Shaoyao Powder(DSP) against mitophagy in rat model of Alzheimer's disease(AD) induced by streptozotocin(STZ) based on PTEN induced putative kinase 1(PINK1)-Parkin signaling pathway. The AD rat model was established by injecting STZ into the lateral ventricle, and the rats were divided into normal group, model group, DSP low-dose group(12 g·kg(-1)·d(-1)), DSP medium-dose group(24 g·kg(-1)·d(-1)), and DSP high-dose group(36 g·kg(-1)·d(-1)). Morris water maze test was used to detect the learning and memory function of the rats, and transmission electron microscopy and immunofluorescence were employed to detect mitophagy. The protein expression levels of PINK1, Parkin, LC3BⅠ/LC3BⅡ, and p62 were assayed by Western blot. Compared with the normal group, the model group showed a significant decrease in the learning and memory function(P<0.01), reduced protein expression of PINK1 and Parkin(P<0.05), increased protein expression of LC3BⅠ/LC3BⅡ and p62(P<0.05), and decreased occurrence of mitophagy(P<0.01). Compared with the model group, the DSP medium-and high-dose groups notably improved the learning and memory ability of AD rats, which mainly manifested as shortened escape latency, leng-thened time in target quadrants and elevated number of crossing the platform(P<0.05 or P<0.01), remarkably activated mitophagy(P<0.05), up-regulated the protein expression of PINK1 and Parkin, and down-regulated the protein expression of LC3BⅠ/LC3BⅡ and p62(P<0.05 or P<0.01). These results demonstrated that DSP might promote mitophagy mediated by PINK1-Parkin pathway to remove damaged mitochondria and improve mitochondrial function, thereby exerting a neuroprotective effect.
本研究基于磷酸酶和张力蛋白同源物缺失性磷酸酶诱导假定激酶1(PINK1)-帕金蛋白信号通路,探讨当归芍药散(DSP)对链脲佐菌素(STZ)诱导的阿尔茨海默病(AD)大鼠模型线粒体自噬的作用机制。通过向侧脑室注射STZ建立AD大鼠模型,并将大鼠分为正常组、模型组、DSP低剂量组(12 g·kg⁻¹·d⁻¹)、DSP中剂量组(24 g·kg⁻¹·d⁻¹)和DSP高剂量组(36 g·kg⁻¹·d⁻¹)。采用Morris水迷宫试验检测大鼠的学习记忆功能,运用透射电子显微镜和免疫荧光法检测线粒体自噬。通过蛋白质免疫印迹法检测PINK1、帕金蛋白、微管相关蛋白1轻链3BⅠ/微管相关蛋白1轻链3BⅡ(LC3BⅠ/LC3BⅡ)和p62的蛋白表达水平。与正常组相比,模型组大鼠学习记忆功能显著降低(P<0.01),PINK1和帕金蛋白表达降低(P<0.05),LC3BⅠ/LC3BⅡ和p62蛋白表达增加(P<0.05),线粒体自噬发生率降低(P<0.01)。与模型组相比,DSP中、高剂量组显著改善了AD大鼠的学习记忆能力,主要表现为逃避潜伏期缩短、在目标象限的停留时间延长和穿越平台次数增加(P<0.05或P<0.01),显著激活线粒体自噬(P<0.05),上调PINK1和帕金蛋白的表达,下调LC3BⅠ/LC3BⅡ和p62的蛋白表达(P<0.05或P<0.01)。这些结果表明,DSP可能通过促进PINK1-帕金蛋白通路介导的线粒体自噬,清除受损线粒体,改善线粒体功能,从而发挥神经保护作用。