【摘要】 目的:探讨针刺对两种不同抑郁模型的起效时间和突触结构的影响。方法:将150只健康成年SPF级C57BL/6J雄性小鼠分为5个时间点组:1小时、3小时、6小时、12小时和24小时。每个时间点组又分为模型组、东莨菪碱组和针刺组,每组10只。在干预前给予小鼠强迫游泳刺激一次,持续时间为15分钟。将120只健康雄性成年SPF级SD大鼠分为3个时间点组:1小时、6小时和24小时。每个时间点组又分为正常组、模型组、东莨菪碱组和针刺组,每组10只。采用慢性不可预测轻度应激(CUMS)与单独培养相结合的方法构建抑郁症大鼠模型。利用SPT和OFT用于评估该模型的成功。在动物的百会(GV20)、印堂(GV29)、合谷(LI4)、太冲(LR3)给予针刺,留针20分钟,期间不进行其他操作。其它组给予相应的药物治疗。根据不同的时间点,对小鼠进行FST检测,对大鼠进行FST和NSFT检测。采用高效液相色谱法检测前额叶皮层和海马单胺类神经递质(NE、DA、DOPAC、5-HT、5-HIAA、HVA)的水平。通过western blot检测前额叶皮层和海马中synapsin I、PSD95、p-mTOR、mTOR和BDNF的表达。结果:根据FST模型小鼠实验部分,在1小时时间点,与模型组比较,针刺组的不动时间减少(P<0.05);其他四个时间点(3小时、6小时、12小时和24小时),东莨菪碱和针刺组的固定时间均减少(P<0.05)。在1小时和24小时时间点,与模型组比较,针刺组前额叶皮层和海马组织的5-HT、5-HIAA、NE、DA、DOPAC和HVA水平无差异(P>0.05);针刺组前额叶皮质synapsinI、PSD95、p-mTOR和BDNF的表达水平升高(P<0.05);针刺组海马synapsinI、PSD95、p-mTOR和BDNF的表达水平差异无差异(P>0.05)。根据CUMS模型大鼠实验部分,在1h、6小时和24小时时间点,与正常组比较,模型组的不动时间和延迟进食时间增加(P<0.05);与模型组比较,针刺组不动时间缩短(P<0.05)。在6小时和24小时时间点,东莨菪碱组不动时间较模型组缩短(P<0.01)。在1小时和24小时,与正常组相比,模型组前额叶皮层和海马组织中5-HT、5-HIAA、NE、DA、DOPAC和HVA的表达水平降低(P<0.05),synapsin I、PSD95、P-mTOR和BDNF的表达水平降低(P<0.01)。与模型组比较,针刺组前额叶皮层和海马synapsin I、PSD95、p-mTOR和BDNF的表达水平升高(P<0.05)。结论:针刺可能通过增加前额叶皮层突触可塑性蛋白的表达来发挥快速抗抑郁作用。更多还原

【Abstract】 Objective:To investigate the effect of acupuncture onset time and synaptic structure in two different models of depression.Methods:A total of 150 healthy male adult SPF C57BL/6J mice were divided into five time-point groups:1 h,3 h,6 h,12 h and 24 h.Each time-point group was further divided into three groups:the model,scopolamine and acupuncture groups with 10 rats each.Rats were given forced swimming stimulation(15 min,once) before the intervention.A total of 200 healthy male adult specific pathogen-free(SPF)Sprague-Dawley(SD) rats were divided into 30 normal rats,and 170 modelling ones.A total of 90 rats were successfully modeled,and were randomly divided into three time-point groups:1 h,6 h and 24 h.And each time-point group was further divided into the normal,model,scopolamine,and acupuncture group.Chronic unpredictable mild stress(CUMS) combined with individual feeding was used to establish the depressed rat model.The sucrose preference test(SPT) and open field test(OFT) were used to evaluate the success of the model.There were 10 rats in each group.The acupuncture intervention was performed once for 20 min on "Baihui(GV20)", "Yintang(GV29)", "Hegu(LI4)", and "Taichong(LR3)" without any other operations for the corresponding animals.Other groups were given corresponding medications.According to different time points,the forced swimming test(FST) was conducted in mice,and the FST and novelty-suppressed feeding test(NSFT) were measured in rats.Monoamine neurotransmitters(NE,DA,DOPAC,5-HT,5-HIAA,HVA) in the prefrontal cortex and hippocampus were detected with high performance liquid chromatography(HPLC).The expression of synapsin I,PSD95,p-mTOR,mTOR,and BDNF in the prefrontal cortex and hippocampus was determined by western blot.Results:At the 1 h time point,compared with the model group,the immobility time was significantly decreased in the acupuncture group(P<0.05).At 3 h,6 h,12 h,and 24 h,compared with the model group,the immobility time was significantly reduced in both scopolamine and acupuncture groups(P <0.05).At the 1 h time point,compared with the normal group,the immobility time of FST and the latency to feed were significantly increased in the model group(P <0.05).Compared with the model group,the immobility time of FST showed a significant reduction in the acupuncture group(P <0.05).At the 6h and 24 h time points,compared with the normal group,the immobility time of FST and the latency to feed were significantly increased in the model group(P <0.05).Compared with the model group,the immobility time of FST was significantly decreased in the scopolamine and acupuncture groups(P <0.05).At the 1 h and 24 h time points,compared with the normal group,the expression levels of 5-HT,5-HIAA,NE,DA,DOPAC,and HVA of the prefrontal cortical and hippocampal tissues were significantly reduced in the model group(P <0.05).At the 1 h and 24 h time points,compared with the model group,the expression levels of synapsin I,PSD95,p-mTOR,and BDNF of the prefrontal cortex were significantly increased in the acupuncture group(P <0.05).At the 1 h and 24 h time points,compared with the normal group,the expression levels of synapsin I,PSD95,p-mTOR,and BDNF of the prefrontal cortex were significantly reduced in the model group(P <0.05).Compared with the model group,the expression levels of synapsin I,PSD95,p-mTOR,and BDNF of the prefrontal cortex were significantly increased in the acupuncture group(P <0.05).At the 1 h and 24 h time points,compared with the normal group,the expression levels of synapsin I,PSD95,p-mTOR,and BDNF of the hippocampal tissues were significantly reduced in the model group(P <0.05).Compared with the model group,the expression levels of synapsin I,PSD95,p-mTOR,and BDNF of the hippocampus were not significantly different in the acupuncture group(P> 0.05).Conclusion:Acupuncture may play a rapid antidepressant effect by increasing the expression of synaptic plasticity proteins in the prefrontal cortex.更多还原