逍遥散对肝郁脾虚证模型大鼠海马和杏仁核AMPA受体亚基基因表达的影响

目的 探讨逍遥散治疗肝郁脾虚证的基因调节机制.方法 75只SD大鼠随机等分为5组,正常组、模型组、假手术组、6-氰基-7-nitroquinoxaline-2,3-二酮(CNQX组)和逍遥散组.以21 d慢性束缚造模型组,在此基础上,运用脑立体定位仪微量注射α-氨基羟甲基恶唑丙酸(AMPA)受体的拮抗剂造CNQX组,加造假手术组为了评估手术创伤对模型的影响程度.逍遥散组造模方法和CNQX组尽可能相似,突出逍遥散和CNQX二者干预的可比性.运用RT-PCR一步法检测海马CA1、CA3和杏仁核AMPA受体的2个重要亚基GluR1 mRNA、GluR2 mRNA的表达变化,比较CNQX组和逍遥散组在各区指标变化趋势是否一致.结果 在海马CA1和CA3区,正常组、CNQX组和逍遥散组间GluR1 mRNA和GluR2 mRNA表达均无统计学意义(P＞0.05);在基底外侧杏仁核BLA区,CNQX组由于拮抗AMPA受体,GIuR1 mRNA和GluR2 mRNA均表达最低,但正常组和逍遥散组间均无统计学意义(P＞0.05).排除了手术创伤等混杂因子,逍遥散组和CNQX组上述2个指标RT-PCR表达结果在海马和杏仁核变化趋势均相似.结论 逍遥散对肝郁脾虚证模型大鼠海马和杏仁核AMPA受体亚基基因表达的调节机制可能和CNQX的调节机制吻合.CNQX通过拮抗杏仁核AMPA受体,抑制杏仁核兴奋,缓解海马各区受损.进而推断逍遥散通过纠正杏仁核和海马的"兴奋一抑制"失衡,重建稳态,来治疗肝郁脾虚证.

Influence of Xiaoyao Powder on subunit gene expression of AMPA acceptor in amygdala and hippocampus in rat model of liver-stagnation and spleen-deficiency syndrome

Objective To investigate the mechanism of gene regulation of xiaoyao Powder in the treatment of liver-stagnation and spleen-deficiency syndrome.Methods All SD rats(n=75)were randomly divided into 5 groups,including the normal group,model group,sham-operation group,CNQX group and Xiaoyao Powder group.The model was established by chronic restraint for 21 days and then CNQX group wag given CNQX microinjection by cerebral stereotaxis instrument.The sham-operation group was set up for review the influence of operative injury on the model.The modeling method of xiaoyao Powder group was as the same ag possible with that of CNQX group for the comparison between these two groups. The expression changes of GluR1-mRNA and GluR2-mRNA were detected, which were two important subunits in CA1 and CA3 regions of hippocampus and AMPA receptor of amygdala, by onestep RT-PCR method. The trends of indexes in all regions were compared between CNQX group and Xiaoyao Powder group. Results The expressions of GluR1-mRNA and GluB2-mRNA had no statistical significance among the normal group, CNQX group and Xiaoyao Powder group in CA1 and CA3 regions of hippocampus (all P＞0.05). In BLA region of amygdala the expressions of GluR1-mRNA and GluR2mRNA reached the lowest in CNQX group because the antagonism of CNQX to AMPA receptor, but had no statistical significance between the normal group and Xiaoyao Powder group ( all P＞0. 05 ). The trends of the expressions of GluRI-mRNA and GluR2-mRNA in hippocampns and amygdala were similar showed by RT-PCR in CNQX group and Xiaoyao Powder group after dismissing confounding factors like operative injury. Conclusion The regulating mechanism of Xiaoyao Powder to the expressions of subunits of hippocampus and amygdala AMPA receptor may be coincidence with that of CNQX in rat model of liver-stagnation and spleen-deficiency syndrome. It is deduced that Xiaoyao Powder cures liver-stagnation and spleen-deficiency syndrome through correcting the disorder of "excitation-inhibition" of hippocampus and amygdala and reconstructing the homeostasis.