针刺对CUMS抑郁大鼠模型行为学及作用机制的研究

发布时间：2018-05-26 07:43

本文选题：针刺 + 抑郁症　；参考：《广州中医药大学》2016年博士论文

【摘要】：目的：通过观察针刺“四神聪”、“内关”、“三阴交”穴对慢性不可预见性轻度应激抑郁模型(CUMS)大鼠体重、饮食以及行为学的影响,并通过观察实验大鼠海马脑区GLU及GABA含量变化、外周血神经递质(DA和5-TH)水平、谷氨酸脱羧酶(GAD65,GAD67)、谷氨酸转运体(EAAT3)、GABA转运体1(GAT1)、NMDA受体亚单位NR2B、GABAA受体的蛋白表达的变化,初步研究针刺抗抑郁的作用原理,并试图探讨其机制。方法：SD大鼠50只,SPF级,雌雄各占一半,体重180-230g,置于模拟日夜的实验室中(白天7：00-19：00,夜晚19：00-7：00),并随机分为空白组、模型组、西药组(氟西汀)、穴位组和非穴组,每组10只。饲养1周后,应用慢性不可预见性轻度应激措施,建立抑郁模型(CUMS),应激措施主要有：禁食、禁水、颠倒昼夜、束缚活动、夹尾、摇晃、足底电击、冰水游泳。除空白对照组外,其余各组大鼠均进行抑郁造模,每天随机选取一种措施进行刺激,每天刺激2次。从入组第1天开始对各组大鼠进行治疗,每日1次,治疗至入组第21天。从入组第1天至第21天,穴位组选取“四神聪”、“内关”、“三阴交”穴进行针刺,每次留针30min,每日1次。非穴组在上述三穴位旁边的非穴位点进行针刺(根据国内外通用非穴位取穴方法,相近两条经络连线中点取为非穴位点),余治疗方法同穴位组。西药组给予氟西汀1.8mg/kg体重,溶于2m1纯净水中灌胃,1次/天。每组动物于入组第1天、第7天、第14天和第21天分别称量体重,记录进食量以及open-field实验,实验结束检测海马脑区GLU及GABA含量、外周血神经递质(DA和5-TH)含量、谷氨酸脱羧酶(GAD65, GAD67)、谷氨酸转运体(EAAT3)、GABA转运体1(GAT1)、NMDA受体亚单位NR2B、GABAA受体的蛋白表达。结果：1、随着造模时间的延长,空白对照组大鼠的体重增加明显,其次为西药组、穴位组体重明显增加；穴位组在后期治疗中增加幅度较西药组较大,但相互之间无明显差异(P0.05)；非穴组体重有所增加,但增加缓慢,增加量也明显比其他各治疗组大鼠少,与模型组之间无明显差异(P0.05)；与空白组相比,各治疗组在治疗21天后体重逐渐接近正常组体重水平,但尚有一定差异(P0.05),与模型组及非穴组相比,差异明显(P0.05),各组蔗糖消耗量及饲料消耗量与体重变化呈正相关。在行为学调节方面：模型组动物与正常组比较,水平运动次数与垂直运动次数均明显减少,差异显著(P0.05)；穴位组、西药组治疗组的水平运动和垂直运动得分同模型组比较,均有显著性(P0.05～0.01),而与正常组相比,则无显著性差异(P0.05)；非穴位组与模型组之间差异不明显(P0.05)；各治疗组间的得分相比无显著差异(P0.05)2、CUMS大鼠模型海马脑区GLU含量明显降低(P0.05),而GABA含量明显升高(P0.05),经过针刺穴位干预,使慢性应激抑郁大鼠模型海马脑区GLU含量明显升高(P0.01),GABA含量明显降低(P0.01)。3、CUMS大鼠模型外周血DA含量较正常组显著降低(P0.05),西药组、穴位组跟模型组比较,差异有统计学意义(P0.01)。各组大鼠外周血5-HT含量空白组与模型组比较,差异有统计学意义(P0.05)；西药组、穴位组跟模型组比较,差异有统计学意义(P0.01)4、CUMS大鼠模型GAD65蛋白表达较正常对照组水平显著升高(P0.01),EAAT3蛋白表达水平显著降低(P0.05),GAT1蛋白表达水平显著升高(P0.05),NR2B蛋白表达水平显著降低(P0.01)；与模型组比较,氟西汀和针刺穴位组GAD65蛋白表达水平显著降低(P0.05),EAAT3蛋白表达水平显著升高(P0.01,P0.05),GAT1蛋白表达水平显著降低(P0.01,P0.05),NR2B蛋白表达水平显著升高(P0.05),而针刺非穴组均无显著差异(P0.05)。结论：1、通过针刺干预,抑郁大鼠蔗糖消耗量、饲料消耗量增加以及体重均增加,同时运动活力增多,由此推断,针刺可缓解抑郁症的躯体及心理功能紊乱。2、针刺的抗抑郁机制与调节多巴胺(DA)、5-TH含量以及兴奋性氨基酸谷氨酸及其受体的表达有关。3、针刺干预抗抑郁作用具有腧穴特异性。
[Abstract]:Objective: To observe the effects of "Si Shen Cong", "Neiguan" and "Sanyinjiao" on the weight, diet and behavior of rats with chronic unpredictable mild stress depression (CUMS), and to observe the changes in the content of GLU and GABA in the hippocampus of rats, the level of peripheral blood neurotransmitters (DA and 5-TH), and glutamic acid decarboxylase (G) (GAD65, G). AD67), glutamate transporter (EAAT3), GABA transporter 1 (GAT1), NMDA receptor subunit NR2B, and the protein expression of GABAA receptor, preliminary study the mechanism of acupuncture antidepressant, and try to explore its mechanism. Methods: 50 SD rats, SPF, female and male, and weight 180-230g, are placed in the Laboratory of day and night (7:00-19:00 in the daytime). At night 19:00-7:00), and randomly divided into blank group, model group, western medicine group (Fu Xiting), acupoint group and non acupoint group, 10 in each group. After 1 weeks of feeding, the chronic unpredictable mild stress measures were used to establish the depression model (CUMS). The stress measures mainly include fasting, water prohibition, reversing day and night, binding activity, tailing, wobble, foot shock, ice water travel In addition to the blank control group, the rest of the rats were all depressive models. Each day was randomly selected to stimulate 2 times a day. From the first days of entering the group, the rats were treated, 1 times a day, and the treatment to the group twenty-first days. From the entry group first to twenty-first days, the point group selected "four Shen Cong", "Neiguan", "Three Yin Jiao". Acupoints were needled with acupuncture at 30min each time, 1 times a day. Non acupoint groups were acupuncturing at the non acupoint points next to the above three acupoints. (according to the common non acupoint acupoint extraction methods at home and abroad, non acupoint points in the middle point of two meridians and collaterals), the residual treatment method was the same as the acupoint group. The western medicine group was given the fluoxetine 1.8mg/kg weight, dissolved in the pure water in 2M1 and 1 times. Each group of animals were weighed on first days, seventh days, fourteenth days and twenty-first days respectively. The content of GLU and GABA in the hippocampus of the hippocampus, the content of neurotransmitters (DA and 5-TH), glutamic acid decarboxylase (GAD67), glutamic acid transporter (EAAT3), GABA transporter 1 (GAT1), NMDA receptor subunit in the hippocampus were measured at the end of the experiment. NR2B, GABAA receptor protein expression. Results: 1, with the prolongation of the molding time, the weight of the blank control group increased significantly, followed by the western medicine group, the acupoint group increased significantly, the acupoint group increased in the later treatment than the western medicine group, but there was no significant difference (P0.05), but the weight of the non acupoint group increased, but increased, but increased, but increased, but increased, but increased, but increased, but increased, but increased, but increased, but increased, but increased, but increased There was no significant difference between the model group and the other treatment group (P0.05). Compared with the blank group, the weight gradually approached the normal weight level in the treatment group after 21 days of treatment, but there was a certain difference (P0.05), and the difference was obvious (P0.05) compared with the model group and the non acupoint group, and the consumption of sucrose and the feeding of each group were significantly different. There was a positive correlation between the amount of material consumption and the change of body weight. In the aspect of behavioral regulation, the number of horizontal movement times and the number of vertical movements in the model group were significantly reduced, and the difference was significant (P0.05). The scores of horizontal and vertical movement in the acupoint group and the western medicine group were significantly higher than those in the model group (P0.05 to 0.01). There was no significant difference in the normal group (P0.05), and there was no significant difference between the non acupoint group and the model group (P0.05); there was no significant difference in the scores between the treatment groups (P0.05) 2, the GLU content in the hippocampus of the CUMS rat model decreased significantly (P0.05), while the GABA content increased significantly (P0.05), and the chronic stress was depressed in the rats through acupuncture point intervention. The content of GLU in the hippocampus of the model was significantly increased (P0.01), the content of GABA decreased significantly (P0.01).3, and the content of DA in the peripheral blood of CUMS rats was significantly lower than that of the normal group (P0.05). The difference was statistically significant (P0.01) compared with the model group (P0.01). The difference was statistically significant (P) in the blank group of peripheral blood 5-HT content in each group (P). 0.05): compared with the model group, the difference between the western medicine group and the model group was statistically significant (P0.01) 4. The expression of GAD65 protein in the CUMS rat model was significantly higher than that in the normal control group (P0.01), the expression level of EAAT3 protein was significantly decreased (P0.05), the expression level of GAT1 protein increased significantly (P0.05), and the expression level of NR2B protein was significantly decreased (P0.01). The expression level of GAD65 protein in fluoxetine and acupuncture point group was significantly decreased (P0.05), the expression level of EAAT3 protein was significantly increased (P0.01, P0.05), the expression level of GAT1 protein was significantly decreased (P0.01, P0.05), the expression level of NR2B protein was significantly increased (P0.05), but there was no significant difference in acupuncture non acupoint group (P0.05). Conclusion: 1, through acupuncture intervention, depression is big. The consumption of sucrose, the increase of feed consumption, the increase of weight and the increase of exercise activity, it is concluded that acupuncture can relieve the physical and psychological dysfunction of depression.2. The antidepressant mechanism of acupuncture is related to the regulation of dopamine (DA), the content of 5-TH and the expression of glutamic acid and its receptors of excitatory amino acids,.3, and acupuncture intervention for antidepressant. The effect has acupoint specificity.
【学位授予单位】：广州中医药大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R245;R-332

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