Zou Ling, Song Xiaoge, Zhao Yanbiao, Qian Tingting, Chu Yifan, Pan Wen, Chu Haoran, Yang Shaojie, Sun Meixiang, Sun Peiyang
Seventh Department of Encephalopathy, Second Affiliated Hospital of Anhui University of CM, Hefei 230061, China.
Research Institute of Acupuncture and Meridian of Anhui University of CM.
Zhongguo Zhen Jiu. 2025 Sep 12;45(9):1281-1289. doi: 10.13703/j.0255-2930.20240922-k0001. Epub 2024 Dec 3.
To observe the effects of acupuncture (for unblocking the obstruction in the governor vessel and regulating the spirit) on depression-like behavior and the hippocampal Endophilin A1/reactive oxygen species (ROS) pathway in the chronic unpredictable mild stress (CUMS) model rats, and explore the mechanism of this therapy for depression.
Forty-eight male SD rats of SPF grade were randomly divided into a normal group (=12) and a modeling group (=36). In the modeling group, CUMS was performed to establish depression model. The successfully-modeled rats were randomized into a model group, a Tongdu Tiaoshen acupuncture group (referred to as the acupuncture group), and a fluoxetine group, with 12 rats in each group. In the acupuncture group, "Baihui" (GV20), "Shenting" (GV24), "Shuigou" (GV26) and "Dazhui" (GV14) were stimulated with acupuncture. This intervention measure was delivered once a day, continuously for 6 days; it was discontinued on day 7 and was completed in 28 days. In the fluoxetine group, intragastric administration was done with fluoxetine solution (2.1 mg/kg), once a day, and for 28 consecutive days. Before and after modeling, and after intervention completion, the body mass, sucrose preference rate and the total distance of movement and the boxes of horizontal crossing in the open field experiment were observed in each group. After intervention, using HE staining, the hippocampal neuron morphology was observed; using Nissl staining, the hippocampal Nissl body number was counted. The hippocampal mitochondria was observed under transmission electron microscopy. The average fluorescence intensity of ROS in hippocampal was determined using flow cytometry. With Western blot method, the protein expression of Endophilin A1, growth associated protein 43 (GAP-43), and brain-derived neurotrophic factor (BDNF) in hippocampal was detected; and with RT-qPCR method, the mRNA expression of Endophilin A1, GAP-43, and BDNF was recorded. Using the immunofluorescence, the average fluorescence intensity of Endophilin A1, GAP-43, and BDNF in hippocampal tissue was determined.
Compared with the normal group, in the model group, the body mass, sucrose preference rate, and the total distance of movement and the boxes of horizontal crossing in the open field experiment decreased (<0.01); the hippocampal neuronal structure was unclear, the matrix was relatively loose, and the number of Nissl body decreased (<0.01); mitochondrial structure was disarranged, the outer membrane was ruptured, mitochondrial cristae was irregular or missed; the average fluorescence intensity of ROS in hippocampal tissue, the protein and mRNA expression and the average fluorescence intensity of Endophilin A1 in hippocampal tissue increased (<0.01), while the protein and mRNA expression of GAP-43 and BDNF and its average fluorescence intensity decreased (<0.01). Compared with the model group, the acupuncture group and the fluoxetine group showed the increase in body mass, sucrose preference rate, the total distance of movement and the boxes of horizontal crossing in the open field experiment (<0.05, <0.01); the hippocampal neuronal structure became relatively clear, the matrix was relatively dense, and the number of Nissl body was elevated (<0.01); mitochondrial structure got normal and disarranged slightly, the average fluorescence intensity of ROS in hippocampal tissue, the protein and mRNA expression and the average fluorescence intensity of Endophilin A1 in hippocampal tissue were reduced (<0.01), while the protein and mRNA expression of GAP-43 and BDNF and the average fluorescence intensity rose (<0.01, <0.05). Compared with the fluoxetine group, the acupuncture group presented the increase in the average fluorescence intensity of ROS, the protein expression and the average fluorescence intensity of Endophilin A1, the protein expression of GAP-43 and the mRNA expression of BDNF (<0.01, <0.05), and the decrease of the protein expression and the average fluorescence intensity of BDNF, the mRNA expression of Endophilin A1, and the average fluorescence intensity of GAP-43 (<0.01, <0.05).
acupuncture alleviates depression-like behaviors in CUMS model rats and protects hippocampal neurons, which may be related to suppressing Endophilin A1/ROS signaling pathway and attenuating oxidative stress reactions.
观察针刺（督脉通阳调神法）对慢性不可预测性温和应激（CUMS）模型大鼠抑郁样行为及海马Endophilin A1/活性氧（ROS）通路的影响，探讨针刺治疗抑郁症的作用机制。
将48只SPF级雄性SD大鼠随机分为正常组（n = 12）和造模组（n = 36）。造模组采用CUMS法建立抑郁模型。将造模成功的大鼠随机分为模型组、通督调神针刺组（简称针刺组）和氟西汀组，每组12只。针刺组针刺“百会”（GV20）、“神庭”（GV24）、“水沟”（GV26）、“大椎”（GV14），每天干预1次，连续6天；第7天停止干预，共干预28天。氟西汀组给予氟西汀溶液（2.1 mg/kg）灌胃，每天1次，连续28天。观察各组造模前后及干预结束后体质量、蔗糖偏爱率、旷场实验总运动距离及水平穿越格数。干预后，采用HE染色观察海马神经元形态；采用尼氏染色计数海马尼氏体数量。透射电镜观察海马线粒体。流式细胞术检测海马组织ROS平均荧光强度。采用蛋白质免疫印迹法检测海马组织中Endophilin A1、生长相关蛋白43（GAP-43）和脑源性神经营养因子（BDNF）的蛋白表达；采用RT-qPCR法检测Endophilin A1、GAP-43和BDNF的mRNA表达。免疫荧光法检测海马组织中Endophilin A1、GAP-43和BDNF的平均荧光强度。
与正常组比较，模型组体质量、蔗糖偏爱率、旷场实验总运动距离及水平穿越格数均降低（P < 0.01）；海马神经元结构不清，基质疏松，尼氏体数量减少（P < 0.01）；线粒体结构紊乱，外膜破裂，线粒体嵴不规则或缺失；海马组织ROS平均荧光强度、Endophilin A1蛋白和mRNA表达及平均荧光强度升高（P < 0.01），而GAP-43和BDNF蛋白及mRNA表达及其平均荧光强度降低（P < 0.01）。与模型组比较，针刺组和氟西汀组体质量、蔗糖偏爱率、旷场实验总运动距离及水平穿越格数均升高（P < 0.05，P < 0.01）；海马神经元结构较清晰，基质致密，尼氏体数量增多（P < 0.01）；线粒体结构基本正常，轻度紊乱，海马组织ROS平均荧光强度、Endophilin A1蛋白和mRNA表达及平均荧光强度降低（P < 0.01），而GAP-43和BDNF蛋白及mRNA表达和平均荧光强度升高（P < 0.01，P < 0.05）。与氟西汀组比较，针刺组ROS平均荧光强度、Endophilin A1蛋白表达及平均荧光强度、GAP-43蛋白表达和BDNF mRNA表达升高（P < 0.01，P < 0.05），BDNF蛋白表达及平均荧光强度、Endophilin A1 mRNA表达和GAP-43平均荧光强度降低（P < 0.01，P < 0.05）。
针刺可改善CUMS模型大鼠的抑郁样行为，保护海马神经元，其机制可能与抑制Endophilin A1/ROS信号通路、减轻氧化应激反应有关。
