摘要
目的探索自拟安神方(简称安神方)对大鼠不同脑区5-羟色胺(5-hydroxytryptamine,5-HT)和多巴胺(dopamine,DA)信号通路的影响,揭示安神方抗大鼠心肾不交证失眠的机制。方法将SD雄性大鼠随机分为正常组,模型组,阳性对照组,安神方高、中、低剂量组,每组10只。除正常组外,其他各组采用多因素方法制备改良的心肾不交证失眠大鼠模型,灌胃治疗2周。HE染色观察大鼠脑组织形态结构。高效液相色谱-电化学法(HPLC-ECD)检测大鼠皮质区和海马区DA和5-HT的含量。免疫蛋白印迹法检测5-HT信号通路上5-HT_(1A)受体、5-HT_(2A)受体、磷酸腺苷反应元件结合蛋白(cAMP-response element binding protein,CREB)、蛋白激酶A(protein kinase A,PKA)和DA信号通路上D_(1)R_(1)在皮质区和海马区的蛋白表达。结果HE染色显示模型组皮质区神经元有少量的凋亡,海马结构区变化不明显;安神方组皮质区神经元凋亡有所减轻。HPLC-ECD结果显示,与正常组比较,模型组皮质区和海马区DA含量均升高(P<0.05),5-HT含量均显著下降(P<0.05);与模型组比较,安神方组皮质区和海马区DA含量显著降低(P<0.05),同时海马区5-HT含量有所提高(P<0.05)。免疫印迹结果显示,与正常组比较,模型组皮质区5-HT_(1A)和CREB蛋白表达降低(P<0.05),海马区D_(1)R_(1)蛋白表达升高(P<0.05);与模型组比较,安神方组皮质区5-HT_(1A)和信号通路上关键蛋白CREB的蛋白表达上调(P<0.05),并且海马区D_(1)R_(1)的蛋白表达下调(P<0.05),其信号通路上CREB蛋白表达差异虽无统计学意义,但各给药组仍有上调趋势。结论安神方抗心肾不交证失眠的机制可能与抑制皮质区神经元凋亡,并促进皮质区和海马区5-HT通路功能、抑制DA通路功能有关。
Objective To reveal the mechanism of Self-prepared Anshen Decoction(自拟安神方,ASD)on insomnia of rats with heart kidney intersection based on the changes of 5-hippocampus and dopamine signaling pathways in cortex and hippocampal.Methods SD male rats were randomly divided into normal group,model group,positive control group and high-dose,medium-dose,low-dose ASD group.Multifactorial stimulus were used to make a modified rat model of insomnia except for the normal group.After 2 weeks,rats in different groups were given different drugs for 2 weeks.HE staining was used to observe morphology change.The content of 5-HT and DA in cortex and hippocampus were detected by high performance liquid chromatography-electrochemical method(HPLC-ECD).Western blotting was used to observe the changes of 5-HT_(1A),5-HT_(2A),D_(1)R_(1),cAMP-response element binding protein(CREB)and protein kinase A(PKA)protein expression in cortex and hippocampus.Results HE staining showed apoptosis of a small amount of neurons in cortex and there were no obvious changes in the hippocampus.ASD group significantly inhibited neuronal apoptosis.HPLC-ECD results showed compared with the normal group,the content of DA was increased(P<0.05)and the content of 5-HT was decreased(P<0.05)in both cortex and hippocampus in model group.Compared with the model group,ASD group significantly decreased the content of DA in cortex and hippocampus(P<0.05),and significantly increased the content of 5-HT in hippocampus(P<0.05).Western blotting results showed compared with the normal group,the expression of 5-HT_(1A) and CREB in cortex were decreased(P<0.05)and the expression of D_(1)R_(1) protein in hippocampus was increased(P<0.05)in model group.Compared with the model group,ASD significantly increased the expression of 5-HT_(1A) and CREB protein in cortex(P<0.05),and significantly decreased the expression of D_(1)R_(1) protein in hippocampus(P<0.05).Although the CREB expression acted in the DA signaling pathway in the hippocampus was not statistically significant,
作者
李雪
徐雅
杜庆红
白俊杰
王旭
李靖
LI Xue;XU Ya;DU Qinghong;BAI Junjie;WANG Xu;LI Jing(Beijing University of Chinese Medicine,Beijing 100029,China;University of Tibetan Medicine,Lhasa 850000,Tibet,China)
出处
《辽宁中医药大学学报》
CAS
2023年第10期20-25,F0003,共7页
Journal of Liaoning University of Traditional Chinese Medicine
基金
西藏自治区藏医药管理局级科技专项项目(JJKT2020006)
西藏自治区科技计划项目(重点研发计划)(XZ202201ZY0019G)
西藏自治区教育厅项目(90020171720022)。
关键词
自拟安神方
心肾不交证失眠
不同脑区
多巴胺
5-羟色胺
Self-prepared Anshen Decoction(自拟安神方)
insomnia with heart kidney intersection
different brain regions
DA
5-HT
