“通督调神”针刺对脑卒中后抑郁大鼠海马神经元保护作用及单胺类神经递质的影响

Protective effects on hippocampal neurons and the influence on hippocampal monoamine neurotransmitters with acupuncture for promoting the circulation of the governor vessel and regulating the mental state in rats with post-stroke depression

目的:观察"通督调神"针刺对卒中后抑郁大鼠海马神经元组织结构以及单胺类神经递质去甲肾上腺素(NE)、5-羟色胺(5-HT)和多巴胺(DA)的影响,探讨"通督调神"针法改善脑卒中后抑郁的可能作用机制。方法:选用清洁级SD大鼠48只,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、非经非穴组采用大脑中动脉闭塞和慢性不可预见性温和性应激法复合制备卒中后抑郁模型;假手术组仅切开皮肤,分离筋膜肌肉,暴露动脉后不进行结扎直接缝合切口。假手术组、模型组不予干预,通督调神组选取"百会""水沟""神庭""大椎"穴,针刺40 min,20 min行针1次,每日针刺1次,每周6次,连续治疗4周。非经非穴组选取大鼠前肢足背侧第3、4跖骨间隙凹陷处为"非经非穴"干预,针刺方法同通督调神组。观察干预后各组大鼠行为学变化,透射电镜观察各组大鼠海马CA1区神经元超微结构,高效液相色谱法测定海马组织中单胺类神经递质NE、5-HT和DA含量。结果:与假手术组比较,模型组大鼠Zea Longa神经行为学评分提高(P<0.01),蔗糖水饮用量降低(P<0.01),敞箱水平运动、垂直运动次数降低(均P<0.01),大鼠海马CA1区神经元超微结构发生明显损伤,海马组织中NE、5-HT和DA含量降低(均P<0.01);与模型组相比较,通督调神组大鼠Zea Longa神经行为学评分降低(P<0.01),蔗糖水饮用量增加(P<0.01),敞箱水平运动、垂直运动次数增多(均P<0.01),大鼠海马CA1区神经元超微结构损害减轻,海马组织中NE、5-HT和DA含量升高(P<0.01,P<0.05);与非经非穴组相比较,通督调神组大鼠Zea Longa神经行为学评分降低(P<0.01),蔗糖水饮用量增加(P<0.01),敞箱水平运动、垂直运动次数增多(均P<0.01),大鼠海马CA1区神经元超微结构损害减轻,海马组织中NE、5-HT和DA含量升高(P<0.01,P<0.05)。结论:"通督调神"针刺能改善卒中后抑郁模型大鼠行为、修复大鼠海马神经元的损伤,可能与其上调海马组织中单胺类神经递质(NE、5-HT、DA)含量有关。

Objective To observe the effects on hippocampal neuronal structure and hippocampal monoamine neurotransmitters such as norepinephrine(NE), 5-hydroxytryptamine(5-HT) and dopamine(DA) with acupuncture for promoting the circulation of the governor vessel and regulating the mental state(Tongdu Tiaoshen acupuncture) in rats with post-stroke depression and explore the underlying effect mechanism of Tongdu Tiaoshen acupuncture on post-stroke depression. Methods A total of 48 clean-grade SD rats were randomized into a sham-operation group, a model group, a Tongdu Tiaoshen acupuncture group and a non-meridian-non-acupoint group, 12 rats in each one. In the model group, the Tongdu Tiaoshen acupuncture group and the non-meridian-non-acupoint group, the compound method of cerebral medial arterial occlusion and the chronic unpredictable mild stress was adopted to prepare the model of post-stroke depression.In the sham-operation group, skin was opened and the fascia muscle was separated, after artery exposed, no ligation was operated and the incision was sutured directly. In the sham-operation group and the model group, no any intervention was performed. In the Tongdu Tiaoshen acupuncture group, acupuncture was applied to "Baihui"(GV 20),"Shuigou"(GV 26),"Shenting"(CV 24) and "Dazhui"(GV 14) for 40 min in each treatment and the needles were manipulated at the acupoints once in 20 min. The treatment was given once a day, 6 times a week for 4 weeks consecutively. In the nonmeridian-non-acupoint group, the depression between the 3 rd and 4 th metatarsal bones on the dorsum of the forelimb was selected in intervention and the needling technique was same as the Tongdu Tiaoshen acupuncture group. The behavioral changes were observed after intervention in each group. Using transmission electron microscopy, the ultrastructure of hippocampal CA 1 neurons was observed in the rats of each group. Using high performance liquid chromatography, the contents of NE, 5-HT and DA were determined. Results Compared with the sham-operation group, the Zea-longa neurobehavioral score was increased(P<0.01), the sucrose water consumption was reduced(P<0.01), the number of times of horizontal movement and vertical movement in open-field test were reduced(both P<0.01), the damage of the ultrastructure of hippocampal CA 1 neurons was aggravated and the contents of NE, 5-HT and DA were reduced in the model group(P<0.01). Compared with the model group, the Zea-Longa neurobehavioral score was reduced(P<0.01), the sucrose water consumption was increased(P<0.01), the number of times of horizontal movement and vertical movement in openfield test were increased(both P<0.01), the damage of the ultrastructure of hippocampal CA 1 neurons was alleviated and the contents of NE, 5-HT and DA were increased in the Tongdu Tiaoshen acupuncture group(P<0.01, P<0.05).Compared with the non-meridian-non-acupoint group, the Zea-Longa neurobehavioral score was reduced(P<0.01),the sucrose water consumption was increased(P<0.01), the number of times of horizontal movement and vertical movement in open-field test were increased(both P<0.01), the damage of the ultrastructure of hippocampal CA 1 neurons was alleviated and the contents of NE, 5-HT and DA were increased in the Tongdu Tiaoshen acupuncture group(P<0.01, P<0.05). Conclusion The Tongdu Tiaoshen acupuncture improves the behavior of the model rats with poststroke depression, repairs hippocampal neuronal damage, which is probably related to the contents of hippocampal monoamine neurotransmitters(NE, 5-HT and DA).