针刺百会、印堂对脑卒中抑郁大鼠神经递质释放及miRNA-219/CaMKⅡγ通路的影响

Effects of Needling CV20 and EX-HN3 on Neurotransmitter Releases and miRNA-219/CaMKⅡγPathway in Rats with Post-Stroke Depression

目的:探讨针刺百会、印堂对脑卒中抑郁(PSD)大鼠神经递质释放及miRNA-219、钙/钙调素依赖性蛋白激酶Ⅱγ(CaMKⅡγ)通路的影响。方法:SD大鼠随机分为假手术组、模型组、针刺组、非经非穴组和文拉法辛组,每组12只,除假手术组外,其余各组建立PSD大鼠模型,分组处理后,各组大鼠进行神经功能缺损评分;以蔗糖水消耗实验及敞箱实验检测各组大鼠抑郁症状;以高效液相色谱法检测各组大鼠脑组织中神经递质去甲肾上腺素(NE)、5-羟色胺(5-HT)和多巴胺(DA)的含量;以实时荧光定量(qRT-PCR)及蛋白免疫印迹法检测各组大鼠脑组织miRNA-219、CaMKⅡγ信号表达情况。结果:与假手术组相比,模型组大鼠神经功能缺损评分、CaMKⅡγmRNA及蛋白表达明显升高(P<0.05),蔗糖水消耗量、水平运动与垂直运动次数、神经递质NE及5-HT、DA含量和脑组织miRNA-219表达明显降低(P<0.05)。与模型组相比,文拉法辛组、针刺组大鼠神经功能缺损评分、CaMKⅡγmRNA及蛋白表达降低(P<0.05),蔗糖水消耗量、水平运动与垂直运动次数、神经递质NE及5-HT、DA含量和脑组织miRNA-219表达升高(P<0.05);非经非穴组大鼠各指标差异无统计学意义(P>0.05)。针刺组与文拉法辛组比较,大鼠各指标差异无统计学意义(P>0.05)。结论:针刺百会、印堂可上调miRNA-219表达,下调CaMKⅡγ表达,修复PSD大鼠神经功能,促进神经递质释放,改善大鼠抑郁症状。

Objective:To investigate the effects of needling Baihui(CV20)and Yintang(EX-HN3)on neurotransmitter releases and miRNA-219/CaMKⅡγ(calcium/calmodulin-dependent protein kinaseⅡγ)pathway in rats with post-stroke depression(PSD).Methods:SD rats were randomly divided into the sham operation group,the model group,the acupuncture group,non-meridian and non-acupoint group and Venlafaxine group,with 12 rats in each group.PSD model was established in each group except for the sham operation group.The scores of neurological deficit were assessed in the groups after corresponding interventions.The depression symptoms were detected by sucrose water consumption experiment and open box experiment.The contents of noradrenaline(NE),5-hydroxytryptamine(5-HT)and dopamine(DA)in brain tissue of rats were detected by high-performance liquid chromatography in the groups.The signal expressions of miRNA-219 and CaMKⅡγin brain tissue of rats in each group were detected by real-time fluorescence quantitative analysis(qRT-PCR)and Western blotting method.Results:Compared to the sham operation group,the scores of neurological deficit,the mRNA and protein expressions of CaMKⅡγwere significantly increased in the model group(P<0.05);whereas the consumption of sucrose water,the times of horizontal and vertical movements,the contents of NE,5-HT and DA,and the expressions of miRNA-219 in brain tissue decreased significantly in the model group(P<0.05).Compared to the model group,the scores of neurological deficit,the mRNA and protein expressions of CaMKⅡγwere significantly decreased(P<0.05);whereas the consumption of sucrose water,the times of horizontal and vertical movements,the contents of NE,5-HT and DA,and the expression of miRNA-219 in brain tissue increased significantly in Venlafaxine group and in the acupuncture group(P<0.05);and there were no significant differences in the indexes of non-meridian and non-acupoint group(P>0.05).There were no significant differences in the indexes between the acupuncture group and Venlafaxine group(P>0.05).Conclusion:Needling CV20 and EX-HN3 can up-regulate the expression of miRNA-219 and down-regulate the expression of CaMKⅡγ,repair the neural function of PSD rats,promote the releases of neurotransmitters and improve the depressive symptoms of the rats.