Sun Yang, Tu Ya, Guo Yu, Jiang Hui-Li, Li Ya-Huan, Wang Yu, Zhao Bing-Cong, Gao Xing-Zhou, Li Xiang, Feng Shi-Xing, Li Yu-Fei
School of Acupuncture-moxibustion and Tuina, Beijing University of Chinese Medicine, Beijing 100029, China; Research Center of Mental and Neurological Disorders, School of Acupuncture-moxibustion and Tuina, Beijing University of Chinese Medicine, Beijing 100029.
Institute of Acupuncture and Moxibustion, China Academy of Chinese Medical Sciences, Beijing 100700.
Zhen Ci Yan Jiu. 2019 Jun 25;44(6):412-8. doi: 10.13702/j.1000-0607.190098.
To investigate the effect of acupuncture on behavior changes and the expression of reactive oxygen species (ROS), cytochrome C, cysteine-containing aspartate-specific proteases-3 (caspase-3), apoptosis inducing factor (AIF) in the hippocampus of rats exposed to chronic restraint stress (CRS), so as to provide experimental evidence for the prevention and treatment of depression.
Thirty two male SD rats were equally randomized into control, model, acupuncture and medication (Fluoxetine) groups according to the random number table. The depression model was established by chronic psychological stress stimulation (fasting, water deprivation, chronic restraint combined with solitary raising for 28 days). Manual acupuncture stimulation (2 circles/s) was applied to "Baihui" (GV20), "Yintang" (GV29) and bilateral "Sanyinjiao" (SP6) for 2 min, followed by retaining the needles for 20 min. Rats in the medication group were treated by gavage of Fluoxetine (0.18 mg/mL, 1 mL/100 g body weight). The treatment was conducted once daily for 28 days. The depression-like behavior was evaluated by using open-field test, sucrose preference test, and body weight. The content of ROS of the right hippocampus tissue was detected by using dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay, and the expression levels of cytochrome C, caspase-3 and AIF proteins of hippocampus were determined by using Western blot.
After 28 days' stress procedure, the depression rats in the model group showed a significant decrease in the numbers of crossing and rearing of open-field test, sucrose preference index and the body weight compared to the control group (<0.01). The content of ROS and expression levels of cytochrome C, caspase-3 and AIF in the hippocampus of the model group were significantly increased relevant to the control group (all <0.01). After the intervention and compared with the model group, the decreased levels of the numbers of crossing and rearing, body weight and sucrose preference index, and the up-regulated levels of content of ROS and expression of cytochrome C, caspase-3 and AIF proteins were all reversed in both acupuncture and medication groups (<0.01，<0.05), displaying an anti-depressant effect. The effect of acupuncture was significantly superior to that of medication in up-regulating the numbers of crossing and rearing (<0.01, <0.05), and in down-regulating the expression of caspase-3 (<0.01). No significant differences were found between the medication and acupuncture groups in increasing the body weight and sucrose preference index, and in down-regulating the expression of cytochrome C and AIF proteins (>0.05).
Acupuncture intervention can significantly reduce ROS content and expression level of cytochrome C, caspase-3 and AIF proteins in the hippocampus possibly by inhibiting the chronic psychological stress-hippocampal oxidative stress-mitochondrial apoptotic pathway in psychological stress-induced depression rats, which may contribute to its effect in relieving depression.
探讨针刺对慢性束缚应激（CRS）大鼠行为变化及海马中活性氧（ROS）、细胞色素C、含半胱氨酸的天冬氨酸蛋白水解酶-3（caspase-3）、凋亡诱导因子（AIF）表达的影响，为抑郁症的防治提供实验依据。
将32只雄性SD大鼠按随机数字表法均分为对照组、模型组、针刺组和药物（氟西汀）组。采用禁食、禁水、慢性束缚结合单笼饲养28天的慢性心理应激刺激建立抑郁症模型。对“百会”（GV20）、“印堂”（GV29）及双侧“三阴交”（SP6）进行手法针刺刺激（2圈/秒），持续2分钟，然后留针20分钟。药物组大鼠灌胃给予氟西汀（0.18mg/mL，1mL/100g体重）。每天治疗1次，共28天。采用旷场试验、蔗糖偏好试验和体重评估抑郁样行为。用二氯二氢荧光素二乙酸酯（DCFH-DA）法检测右侧海马组织中ROS的含量，用蛋白质免疫印迹法检测海马中细胞色素C、caspase-3和AIF蛋白的表达水平。
应激28天后，与对照组相比，模型组抑郁大鼠旷场试验中的穿越次数、直立次数、蔗糖偏好指数及体重均显著降低（<0.01）。与对照组相比，模型组海马中ROS含量及细胞色素C、caspase-3和AIF的表达水平均显著升高（均<0.01）。干预后，与模型组相比，针刺组和药物组的穿越次数、直立次数、体重及蔗糖偏好指数降低水平，以及ROS含量及细胞色素C、caspase-3和AIF蛋白表达上调水平均得到逆转（<0.01，<0.05），显示出抗抑郁作用。在增加穿越次数和直立次数方面（<0.01，<0.05），以及下调caspase-3表达方面（<0.01），针刺组的效果显著优于药物组。在增加体重和蔗糖偏好指数，以及下调细胞色素C和AIF蛋白表达方面，药物组与针刺组之间无显著差异（>0.05）。
针刺干预可能通过抑制心理应激诱导的抑郁症大鼠慢性心理应激-海马氧化应激-线粒体凋亡途径，显著降低海马中ROS含量及细胞色素C、caspase-3和AIF蛋白的表达水平，这可能是其缓解抑郁的作用机制。
