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  • Title: [Effect of Tongdu Tiaoshen acupuncture on PI3K/Akt/mTOR signaling pathway and autophagy-related proteins of hippocampus in rats with post-stroke depression].
    Author: Sun PY, Li PF, Wang T, Wu J, Li N, Liu H, Li AH, Zhang JJ, Chu HR.
    Journal: Zhongguo Zhen Jiu; 2020 Nov 12; 40(11):1205-10. PubMed ID: 33788489.
    Abstract:
    OBJECTIVE: To observe the improvement of Tongdu Tiaoshen acupuncture on depression-like behavior in rats with post-stroke depression (PSD), and to explore its possible mechanism. METHODS: A total of 48 SD rats were randomized into a sham-operation group, a model group, a Tongdu Tiaoshen group and a non-meridian-non-acupoint group, 12 rats in each group. In the the model group, the Tongdu Tiaoshen group and the non-meridian-non-acupoint group, PSD model was established by the compound method of cerebral medial arterial occlusion, chronic unpredictable mild stress and isolation. Acupuncture was applied at "Dazhui"(GV 14), "Shuigou"(GV 26), "Baihui"(GV 20) and "Shenting" (GV 24) in the Tongdu Tiaoshen group, and at the bilateral dorsum hollows between 3rd and 4th metatarsal bones of the forelegs in the non-meridian-non-acupoint group, needles were retained for 40 min in the both groups, once a day, 6 times a week for 4 weeks. Before model establishment and after intervention, the ethology indice of Zea Longa neurological behavioral score, open-field test and sucrose preference test were evaluated in the 4 groups. The levels of phosphatidylinositol 3-kinase (PI3K), phosphorylated protein kinase B (p-Akt), phosphorylated mammalian target of rapamycin (p-mTOR), Beclin1 and LC3B-Ⅱ/Ⅰin the CA1 region of hippocampus were detected by western blot. The expressions of Beclin1, LC3B-ⅡmRNA in the CA1 region of hippocampus were detected by real-time PCR. RESULTS: After model established, compared with the sham-operation group, the Zea Longa neurological behavioral scores were increased (P<0.05), the scores of horizontal and vertical motion of the open-field test and the consumption of sucrose solution were decreased in the model group, the Tongdu Tiaoshen group and the non-meridian-non-acupoint group (P<0.05). After intervention, compared with the sham-operation group, the Zea Longa neurological behavioral score, the expression of Beclin1, LC3B-Ⅱ/Ⅰ and LC3B-Ⅱin the CA1 region of hippocampus were increased (P<0.05), the scores of horizontal and vertical motion of the open-field test, the consumption of sucrose solution and the expression of PI3K, p-Akt and p-mTOR were decreased in the model group (P<0.05). After intervention, compared with the model group and the non-meridian-non-acupoint group, the Zea Longa neurological behavioral score, the expression of Beclin1, LC3B-Ⅱ/Ⅰ and LC3B-Ⅱin the CA1 region of hippocampus were decreased (P<0.05), the scores of horizontal and vertical motion of the open-field test, the consumption of sucrose solution and the expression of PI3K, p-Akt and p-mTOR were increased in the Tongdu Tiaoshen group (P<0.05). CONCLUSION: Tongdu Tiaoshen acupuncture can effectively improve the depression-like behavior in PSD rats, its mechanism may relate to the activation of PI3K/Akt/mTOR signaling pathway and the inhibition of hippocampal neuron autophagy. 目的：观察“通督调神”针刺对脑卒中后抑郁（PSD）模型大鼠抑郁症状的改善作用，并探讨其可能作用机制。方法：将48只SD大鼠随机分为假手术组、模型组、通督调神组和非经非穴组，每组12只。模型组、通督调神组和非经非穴组大鼠采用大脑中动脉线栓阻塞法制备脑卒中模型，采用慢性不可预知性温和性应激法后孤养法复合制备PSD模型。通督调神组针刺“大椎”“水沟”“百会”“神庭”，非经非穴组大鼠针刺双侧前肢足背侧第3、4跖骨间隙凹陷处，两组均留针40 min，每日针刺1次，每周6 d，连续干预4周。各组大鼠分别于造模后与干预结束后进行行为学指标检测（Zea Longa神经行为学评分、敞箱实验、蔗糖水消耗实验）。Western blot法检测大鼠海马CA1区磷脂酰肌醇-3-激酶（PI3K）、磷酸化蛋白激酶B（p-Akt）、磷酸化哺乳动物雷帕霉素靶蛋白（p-mTOR）、Beclin1、LC3B-Ⅱ/Ⅰ表达水平，real-time PCR法检测大鼠海马CA1区Beclin1、LC3B-ⅡmRNA的表达。结果：造模后，模型组、通督调神组、非经非穴组大鼠Zea Longa神经行为学评分均高于假手术组（P<0.05），敞箱实验水平与垂直运动积分、蔗糖水消耗量均低于假手术组（P<0.05）。干预后，模型组大鼠Zea Longa神经行为学评分，海马CA1区Beclin1、LC3B-Ⅱ/Ⅰ、LC3B-Ⅱ表达水平高于假手术组（P<0.05）；敞箱实验水平与垂直运动积分、蔗糖水消耗量，海马CA1区PI3K、p-Akt、p-mTOR表达水平低于假手术组（P<0.05）。干预后，通督调神组大鼠Zea Longa神经行为学评分，海马CA1区Beclin1、LC3B-Ⅱ/Ⅰ、LC3B-Ⅱ表达水平低于模型组及非经非穴组（P<0.05），敞箱实验水平与垂直运动积分、蔗糖水消耗量，海马CA1区PI3K、p-Akt、p-mTOR表达水平高于模型组及非经非穴组（P<0.05）。结论：“通督调神”针刺能明显改善PSD大鼠抑郁症状，其作用机制可能与激活PI3K/Akt/mTOR通路，抑制海马神经元自噬有关。.
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