Li Zirong, Shu Yi, Liu Deguo, Xie Sheng, Xian Liangbo, Luo Jiaqi, Huang Xiuwen, Jiang Haixing
The First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning, China.
Graduate School, Guangxi University of Chinese Medicine, Nanning, China.
Brain Res Bull. 2025 Jan;220:111141. doi: 10.1016/j.brainresbull.2024.111141. Epub 2024 Dec 3.
Using a chronic unpredictable mild stress (CUMS) combined with multi-platform water environment sleep deprivation (SD) as an animal model, the occurrence and development of human depression combined with insomnia were simulated. The abnormal mitochondrial autophagy signaling caused by the putative kinase 1/Parkin E3 ubiquitin protein ligase (Pink1/Parkin) signaling pathway directly affects the normal secretion of melatonin by the pineal gland, which may explain the pathogenesis of depression combined with insomnia. This study aims to explore the depression-like behavior, sleep changes, central oxidative stress response, pineal mitochondrial autophagy damage, melatonin secretion, histopathological changes of the pineal gland, and the expression of Pink1/Parkin signaling-related factors in CUMS+SD rats. The results showed that the levels of reactive oxygen species (ROS) in cerebrospinal fluid of CUMS+SD rats significantly increased along with the inflammatory factors Interleukin-1β (IL-1β) and nuclear factor kappa-B (NF-κB) in cerebrospinal fluid. In addition, the number of pineal gland cells significantly decreased, cell boundaries became blurred, cell volume shrank, and apoptotic bodies appeared in the pineal gland tissue under HE staining, indicating pineal gland inflammation. Sleep deprivation further disrupted the levels of autophagy damage factors, including histamine (MDA), glutathione (GSH), and catalase (CAT), in the cerebrospinal fluid of CUMS+ SD rats. Transmission electron microscopy of the pineal gland in CUMS+SD rats revealed damage to mitochondrial autophagy. The levels of 5-hydroxytryptamine (5-HT) and aromatic amine-N-acetyltransferase (AANAT) in the cerebrospinal fluid, as well as melatonin levels in the pineal gland, were significantly decreased. Additionally, the expression of IL-1β, NF-κB, Pink1, and Parkin in the pineal gland of CUMS+SD rats significantly increased. The expression of microtubule-associated protein 1 light chain 3-β (LC3), selective autophagy adaptor protein (P62), cytochrome c oxidase IV (COXIV), and mitochondrial outer membrane translocation enzyme 20 (TOM20) proteins downstream of the Pink1/Parkin signaling pathway was enhanced, while the expression of downstream brain-derived neurotrophic factor (BDNF), Beclin 1, and BCL2 interacting protein 3 (BNIP3) proteins was negatively regulated. Pink1/Parkin signaling may specifically respond to mitochondrial autophagy damage in the pineal gland, affecting the normal synthesis and secretion of melatonin in the pineal gland. In summary, mitochondrial autophagy damage in the pineal gland affects the normal secretion of melatonin in CUMS+SD rats, which is closely related to the specific autophagy signaling impairment of Pink1/Parkin pathway, which may mediate the occurrence of depression combined with insomnia.
以慢性不可预测性温和应激（CUMS）联合多平台水环境睡眠剥夺（SD）作为动物模型，模拟人类抑郁合并失眠的发生发展过程。由假定激酶1/帕金E3泛素蛋白连接酶（Pink1/帕金）信号通路引起的线粒体自噬信号异常，直接影响松果体褪黑素的正常分泌，这可能解释了抑郁合并失眠的发病机制。本研究旨在探讨CUMS+SD大鼠的抑郁样行为、睡眠变化、中枢氧化应激反应、松果体线粒体自噬损伤、褪黑素分泌、松果体组织病理学变化以及Pink1/帕金信号相关因子的表达。结果显示，CUMS+SD大鼠脑脊液中活性氧（ROS）水平显著升高，同时脑脊液中的炎性因子白细胞介素-1β（IL-1β）和核因子κB（NF-κB）水平也显著升高。此外，HE染色显示松果体组织中细胞数量显著减少，细胞边界模糊，细胞体积缩小，出现凋亡小体，提示松果体发生炎症。睡眠剥夺进一步扰乱了CUMS+SD大鼠脑脊液中自噬损伤因子的水平，包括组胺（MDA）、谷胱甘肽（GSH）和过氧化氢酶（CAT）。透射电子显微镜观察发现CUMS+SD大鼠松果体线粒体自噬受损。脑脊液中5-羟色胺（5-HT）和芳香胺-N-乙酰基转移酶（AANAT）水平以及松果体中褪黑素水平均显著降低。此外，CUMS+SD大鼠松果体中IL-1β、NF-κB、Pink1和帕金的表达显著增加。Pink1/帕金信号通路下游的微管相关蛋白1轻链3-β（LC3）、选择性自噬衔接蛋白（P62）、细胞色素c氧化酶IV（COXIV）和线粒体外膜转位酶20（TOM20）蛋白的表达增强，而下游脑源性神经营养因子（BDNF）、Beclin 1和BCL2相互作用蛋白3（BNIP3）蛋白的表达受到负调控。Pink1/帕金信号可能特异性地响应松果体中的线粒体自噬损伤，影响松果体中褪黑素的正常合成和分泌。综上所述，松果体线粒体自噬损伤影响CUMS+SD大鼠褪黑素的正常分泌，这与Pink1/帕金通路特异性自噬信号受损密切相关，可能介导了抑郁合并失眠的发生。
