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目的:探讨逍遥散治疗肝郁脾虚证的基因调节机制。方法:25只SD雄性大鼠随机等分为5组,正常组、模型组、假手术组、α-氨基羟甲基恶唑丙酸(AMPA)组和逍遥散组。以21d慢性束缚应激方法造肝郁脾虚证模型组。假手术组、AMPA组和逍遥散组均采用21d慢性束缚联合脑部埋管微量注射方法造模,AMPA组在双侧海马CA1区埋管微量注射AMPA,逍遥散组灌服逍遥散溶液。运用RT-PCR一步法检测海马CA1区和杏仁核区AMPA受体的重要亚基GluR1 mRNA和GluR2 mRNA的表达变化。结果:在杏仁核区,与AMPA组比较,逍遥散组GluR1mRNA和GluR2 mRNA表达差异无统计学意义;在海马CA1区,与正常组比较,AMPA组GluR1 mRNA和GluR2mRNA表达差异均有统计学意义(P<0.01,P<0.05);与AMPA组比较,逍遥散组GluR1 mRNA和GluR2 mRNA表达差异有统计学意义(P<0.01)。结论:结合前期实验,从基因表达角度,进一步推断逍遥散通过纠正杏仁核和海马AMPA受体的“兴奋-抑制”失衡,重建稳态,来治疗肝郁脾虚证。
Objective: To investigate the gene regulation mechanism of Xiaoyao powder in treatment of liver depression and spleen deficiency syndrome. Methods: Twenty-five SD male rats were randomly divided into five groups: normal group, model group, sham operation group, AMPA group and Xiaoyao powder group. 21d chronic restraint stress method to make the model of liver depression and spleen deficiency model group. Sham-operated group, AMPA group and Xiaoyao powder group were all treated with 21-day chronic tethering and brain-embedded microinjection method. AMPA group was injected with AMPA in bilateral CA1 area of hippocampus, Xiaoyao powder group was given Xiaoyaosan solution. The expression of GluR1 mRNA and GluR2 mRNA, which are important subunits of AMPA receptor in hippocampal CA1 region and amygdala, were detected by RT-PCR one-step method. Results: Compared with AMPA group, there was no significant difference in GluR1mRNA and GluR2 mRNA expression in AMPA group. In hippocampal CA1 region, the expression of GluR1 mRNA and GluR2 mRNA in AMPA group were significantly different P <0.01, P <0.05). Compared with AMPA group, the expression of GluR1 mRNA and GluR2 mRNA in Xiaoyaosan group had statistical significance (P <0.01). Conclusion: Combined with the previous experiments, from the perspective of gene expression, Xiaoyao Powder further concludes that Xiaoyao San can treat liver-spleen-deficiency syndrome by correcting the “excitement-inhibition” imbalance of amygdala and hippocampal AMPA receptors and rebuilding the homeostasis.