电针对慢性应激抑郁大鼠脑海马磷酸二酯酶-4亚型表达的影响

发布时间：2018-08-11 20:04

【摘要】：抑郁症是一种综合症状的情感障碍性精神疾病,其特征为情绪上的长期低落。其有较高概率的发病、致残和自杀比例,并有情绪低落、焦虑、精神活动受损、食欲下降和失眠等特点。因而成为一种极大损害人类健康的疾病。据世界卫生组织WHO公开的数据统计显示,抑郁症被列为全球上高居第四位的疾病。全球抑郁症的平均发病率保守估计约为11%,预计到2020年,即五年内,将成为仅次于心脏病的第二大疾病隐患。在中国,抑郁症发病率约为3%～5%,目前有超过2600余万人因抑郁症饱受困扰。最近的流行病学调查表明,抑郁症的终生患病率可达到为16%,排名在中国疾病负担第二高的疾病。目前尚缺少特异性的方法在诊断学上为抑郁提供完美的解决方案。治疗方法以西药的治疗及电休克疗法为主。针刺疗法对治疗抑郁症的治疗在安全性、有效性上都有很好的表现,且与西医抗抑郁药相比,针刺拥有起效迅速、毒副作用相对极小、费用相比较明显低廉等一系列优势。针刺治疗抑郁的作用机制已有大量的研究成果,但还未能完全阐明其起效的靶点及全部通路,尚需开展进一步的研究。目的本研究通过电针对慢性应激抑郁模型大鼠行为学观察结果的影响,并综合运用分子生物学方法进一步探讨电针治疗抑郁症的机制,试图阐明电针对治疗抑郁症潜在靶标PDE4A、PDE4D表达的影响。方法1.动物分组用随机方法将动物分成四组,每组8只,分组分别为:空白组、模型对照组、药物对照组、电针治疗组。各组大鼠在实验前喂养一周,以适应环境。然后开始慢性应激刺激和不同干预,空白组大鼠依然用不分笼群养模式,其余组大鼠每笼1只孤养方式。空白组与其余各组大鼠饲养于隔离的不同饲养室。2.造模方法采用慢性应激模型结合孤养,参考文献方法,进行慢性应激郁症大鼠模型的制备。造模动物模型对照组、药物对照组、电针治疗组孤养,接受21天7种不同的应激刺激,包括冷水游泳、禁水、昼夜颠倒、禁食、夹尾、束缚、潮湿环境,每日随机安排一种刺激,每种刺激平均3次。3.处理方法:空白组:在正常饲养条件下21天,采用非孤养模式,期间不予应激刺激;模型对照组..慢性应激造模21天,并在21天内每日进行与电针组等同的束缚干预,时间长度相同,强度同等;药物对照组:慢性应激加孤养,每日配药,按1mg/1ml的比例将百忧解以蒸馏水配制,按照2mg/kg体重,灌胃途径给药,给药后1h应激造模,每天一次,共21天;电针治疗组:慢性应激与孤养结合,电针治疗21天,并于每天电针1h后进行应激造模。4.检测指标及方法:采用旷场实验(Open—field Test)、体质量(Body Weight)和糖水偏好实验(Sucrose Preference Test)评价模型大鼠行为学改变情况,运用Real-Time PCR技术检测大鼠海马组织PDE4A、PDE4DmRNA的含量,使用蛋白质印迹法(Western Blot)检测海马组织PDE4A、PDE4D的蛋白表达水平。结果1.行为学变化实验前各组大鼠行为学表现均无统计学差异(p0.05)。实验结束后,体重、糖水消耗量及糖水偏好百分率、旷场实验水平穿越格数、垂直竖立次数空白组均高于模型组,有显著性差异(P0.01);药物组和电针治疗组大鼠在以上方面,均高于与模型组,具有显著性差异(P0.01);糖水消耗量方面,与药物组相比,电针组大鼠的糖水消耗量没有显著性差异(p0.05);与糖水偏好百分率上,与药物组相比,电针组大鼠的糖水偏好百分率高于药物组,有统计学差异(p0.05)。2.海马组织中PDE4A、PDE4D mmRNA表达PDE4A、PDE4DmRNA的含量,与空白组比较,模型组大鼠海马表达的相对含量明显增多(P0.01),而电针组大鼠海马表达相对含量与模型组比较明显减少(P0.01),且与空白组比较无统计学差异(P0.05)。与空白组比较,药物组大鼠海马表达相对含量无统计学差异(P0.05)。3.海马组织中PDE4A、PDE4D蛋白相对表达含量PDE4A、PDE4D蛋白相对表达含量上,与空白组比较,模型组大鼠海马蛋白表达相对含量明显增多(P0.01),药物组的表达也增多,有统计学差异(P0.05);电针组与药物组在表达上有统计学差异,P0.01。结论1、氟西汀与电针的干预可改善抑郁症状,电针在改善快感缺失行为上有更明显的优势,可以提高对于奖励机制的反应敏感性。2、PDE4A和PDE4D与抑郁症存在着很大程度的关联。3、电针可以通过抑制PDE4亚型中的PDE4A、PDE4DmRNA的表达来发挥其抗抑郁作用。可产生类似PDE4抑制剂对于抑郁症的改善效果,而不涉及PDE4抑制剂的副作用。4、电针可能是通过抑制PDE4的与抑郁相关的几个单一亚型,更专一地调控与抑郁相关的靶点,进而更好地发挥治疗抑郁效果,这是其改善抑郁状态的可能作用机制之一。
[Abstract]:Depression is a psychiatric disorder characterized by a combination of emotional symptoms characterized by long-term emotional depression. It has a high incidence of morbidity, disability and suicide, and has the characteristics of depression, anxiety, impaired mental activity, loss of appetite and insomnia. It has become a disease that greatly damages human health, according to the World Health Organization WH. Depression is ranked the fourth highest disease in the world, according to O. The global average incidence of depression is conservatively estimated to be about 11%. It is expected to be the second largest disease risk after heart disease in 2020, i.e. within five years. In China, the incidence of depression is about 3%-5%. More than 26 million people are suffering from depression. Recent epidemiological studies have shown that the lifetime prevalence of depression can reach 16%, ranking second highest in China's disease burden. Currently, there is no specific method to provide a perfect diagnostic solution for depression. Compared with Western antidepressants, acupuncture has a series of advantages, such as rapid onset, relatively minimal toxic and side effects, and relatively low cost. There have been a lot of research results on the mechanism of acupuncture in treating depression, but the target of its effect has not been fully clarified. Objective To study the effect of Electroacupuncture on behavioral observation of chronic stress depression model rats, and to explore the mechanism of electroacupuncture in treating depression by using molecular biology methods, so as to clarify the effect of Electroacupuncture on the expression of PDE4A and PDE4D. Methods 1. Animals were randomly divided into four groups: blank group, model control group, drug control group and electroacupuncture treatment group. Rats in each group were fed for one week before the experiment to adapt to the environment. Rats in blank group and other groups were fed in separate feeding rooms. Stimulation, including cold water swimming, water prohibition, day and night inversion, fasting, tail clipping, restraint, wet environment, a daily randomized stimulus, each stimulus averaged 3.3. Processing methods: blank group: 21 days under normal feeding conditions, using non-solitary mode, during which no stress stimulation; model control group.. Chronic stress model 21 days, and 21 days a day within 21 days a day. Drug control group: chronic stress plus orphanage, daily prescription, according to the proportion of 1 mg / 1 ml Prozac with distilled water, according to 2 mg / kg body weight, gastric administration, 1 hour after the administration of stress modeling, once a day for 21 days; Electroacupuncture treatment group: chronic stress and orphanage All rats were treated with electroacupuncture for 21 days, and the stress model was made after electroacupuncture for 1 hour every day. 4. Detection indexes and methods: The behavior changes of model rats were evaluated by open-field test, body weight and sucrose preference test, and the expression of PDE4A and PDE4D mRNA in hippocampus was detected by Real-Time PCR. Western Blot was used to detect the expression of PDE4A and PDE4D in the hippocampus. Results 1. There was no significant difference in the behavior of the rats before and after the experiment (p0.05). After the experiment, body weight, sugar consumption and sugar preference percentage, horizontal crossing lattice number, vertical erection times in open field experiment were observed. Compared with the model group, there was no significant difference in sugar consumption between the electroacupuncture group and the drug group (p0.05); and there was no significant difference in sugar consumption between the electroacupuncture group and the drug group (p0.05). Compared with the blank group, the relative content of PDE4A and PDE4D mRNA in the hippocampus of the model group increased significantly (P 0.01), while the relative content of PDE4A and PDE4D mRNA in the hippocampus of the electroacupuncture group was higher than that of the model group (P 0.05). Compared with the blank group, the relative expression of PDE4A and PDE4D protein in the hippocampus of the drug group was not significantly different (P 0.05). 3. The relative expression of PDE4A and PDE4D protein in the hippocampus of the model group was relative to that of the blank group. The content of fluoxetine was significantly increased (P 0.01), and the expression of drug group was also increased (P 0.05); the expression of fluoxetine was significantly different from that of drug group (P 0.01). Sex. 2, PDE4A and PDE4D are significantly associated with depression. 3. Electroacupuncture can exert its antidepressant effect by inhibiting the expression of PDE4A and PDE4D mRNA in PDE4 subtypes. It can produce an improvement effect similar to PDE4 inhibitors on depression, but does not involve the side effects of PDE4 inhibitors. 4. Electroacupuncture may inhibit PDE4 and depression. One of the possible mechanisms by which depression can be improved is that the related single subtypes more specifically regulate the targets associated with depression and thus play a better role in the treatment of depression.
【学位授予单位】：北京中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R245

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