丹栀逍遥散对广泛性焦虑大鼠Papez环路内神经营养因子及受体表达的影响

Effect of Danzhi Xiaoyao Powder on Expression of Neurotrophic Factors and Receptors in Papez Circuit of Generalized Anxiety Rats

目的观察中药丹栀逍遥散对广泛性焦虑(GAD)大鼠脑Papez环路上神经营养因子[脑源性神经营养因子(BDNF)、碱性成纤维生长因子(bFGF)、神经生长因子(NGF)]及脑源性神经营养因子受体(TrkB)、碱性成纤维生长因子受体(bFGFR)表达的影响。方法将雄性Wistar大鼠随机分为正常组、模型组、中药组(丹栀逍遥散组)和西药组(盐酸丁螺环酮组),每组8只。模型组、中药组、西药组采用天敌暴露饮水冲突刺激法建立GAD大鼠模型。中药组和西药组分别以丹栀逍遥散和盐酸丁螺环酮水溶液灌胃,每天1次,连续21 d。正常组和模型组蒸馏水灌胃21 d。造模成功后,取脑组织,行石蜡切片,免疫组织化学SP法检测Papez环路内海马(CA1、CA2、CA3、DG区)和杏仁核、丘脑前核的BDNF、TrkB、bFGF、bFGFR及NGF的表达。结果与正常组比较,模型组海马各区和杏仁核区TrkB、NGF的表达水平明显降低(P<0.05或P<0.01);模型组海马各区bFGF、bFGFR的表达水平明显升高,杏仁核区的表达水平明显下降(P<0.05或P<0.01);模型组海马各区及杏仁核区、丘脑前核区的BDNF表达与其他3组间比较,差异均无统计学意义(P>0.05),但海马CA1区模型组与正常组有下降趋势。与模型组比较,中药组海马各区和杏仁核区bFGF、TrkB、NGF的表达水平明显升高(P<0.05或P<0.01);中药组海马各区和杏仁核区bFGFR的表达水平明显升高,西药组海马各区和bFGFR的表达水平明显降低(P<0.05);丘脑前核区的表达在模型组与正常组以及中药组与西药组之间的比较,差异均无统计学意义(P>0.05)。结论经过中药丹栀逍遥散干预的GAD大鼠,海马和杏仁核区内bFGF与其受体bFGFR,BDNF的受体TrkB和NGF的表达水平明显回升,有利于bFGF、BDNF、NGF更好地发挥其生理功能,促进损伤脑区局部微环境的恢复,增强神经可塑性,保护神经元,抑制凋亡与坏死,促进神经的修复以及神经干细胞的增殖与分化。提示疏肝清热健脾法能够通过刺激神经营养因子在海马及杏仁核内的表达来发挥其促神经修复功能。

Objective To observe the expression of neurotrophic factors(BDNF,bFGF,NGF)and their receptors(TrkB,Bfgfr)in the Brain Papez circuit of rats with generalized anxiety disease(GAD)and the effect of Danzhi Xiaoyao Powder on them.Methods Male wistar rats were randomly divided into normal group,model group,traditional Chinese medicine group(Danzhi Xiaoyao Powder group)and Western medicine group(buspirone hydrochloride group),with 8 rats in each group.GAD rat models were established by natural enemy exposure and drinking water conflict stimulation in model group,traditional Chinese medicine group,and western medicine group.The rats in traditional Chinese medicine group,and western medicine group were given Danzhi Xiaoyao Powder and buspirone hydrochloride solution by gavage once a day for 21 days.The normal group and model group were given distilled water by gavage for 21 days.The expression of BDNF,TrkB,bFGF,bFGFR,and NGF in hippocampus(CA1,CA2,CA3,DG)and amygdala,anterior thalamic nucleus were detected by immunohistochemistry SP method.Results Compared with the normal group,the expression levels of TrkB and NGF in hippocampus and amygdala of model group were significantly decreased(P<0.05 or P<0.01).In the model group,the expression levels of bFGF and bFGFR in hippocampus were significantly increased,while those in amygdala were significantly decreased(P<0.05 or P<0.01).There were no significant difference in BDNF expression among the model group and the other three groups,but there was a downward trendency between the model group and the normal group in hippocampus CA1.Compared with the model group,the expression levels of bFGF,TrkB and NGF in hippocampus and amygdala of traditional Chinese medicine group were significantly increased(P<0.05 or P<0.01).The expression level of bFGFR in hippocampus and amygdala of traditional Chinese medicine group was significantly increased,and bFGFR in hippocampus that was significantly decreased in western medicine group(P<0.05).There was no significant difference in the expression of the anterior thalamic nucleus between 4 groups.Conclusion After intervention of Danzhi Xiaoyao Powder,the expression levels of bFGF,bFGFR,TrkB,and NGF in hippocampus and amygdala of GAD rats are increased significantly,to promote bFGF,BDNF,and NGF physiological functions,and promote the recovery of local microenvironment,enhance neural plasticity,protect neurons,inhibit apoptosis and necrosis,promote the nerve repairment and NSCs proliferation and differentiation.The result suggests that Shugan Qingre Jianpi method can promote nerve repair by stimulating the expression of neurotrophic factor in hippocampus and amygdala.