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目的:观察肝郁脾虚证模型大鼠海马色氨酸羟化酶2(TPH2)与吲哚胺-2,3-双加氧酶1(IDO1)表达的变化并探讨逍遥散的调节作用。方法:雄性SD大鼠随机分成4组,分别是正常组、模型组、逍遥散组、氟西汀组,每组6只。采用慢性束缚应激的方法制备大鼠肝郁脾虚证模型,造模持续21 d。通过荧光定量PCR与Western-blot方法检测各组大鼠海马TPH2与IDO1的表达情况。结果:模型组大鼠海马TPH2 mRNA的表达水平低于其余3组大鼠(P<0.05),逍遥散与氟西汀对TPH2 mRNA的表达有一定的上调作用;模型组大鼠海马IDO1 mRNA的表达显著增加(P<0.01),逍遥散与氟西汀对IDO1 mRNA表达的下调作用明显(P<0.01),且逍遥散对IDO1 mRNA的下调作用更明显。模型组、逍遥散组、氟西汀组大鼠海马TPH2的蛋白的表达低于正常组(P<0.01),逍遥散组TPH2蛋白表达高于模型组(P<0.05);模型组大鼠海马IDO1的蛋白表达显著高于其余3组(P<0.01)。结论:肝郁脾虚证模型大鼠海马TPH2的表达减少,IDO1的表达增多,逍遥散可能通过调节海马TPH2与IDO1的表达水平进而影响5-HT的含量,起到治疗作用。
Objective: To observe the changes of hippocampal tryptophan hydroxylase 2 (TPH2) and indoleamine-2,3-dioxygenase 1 (IDO1) in rats with liver-qi and spleen-deficiency syndrome and to explore the regulatory effect of Xiaoyaosan. Methods: Male SD rats were randomly divided into 4 groups: normal group, model group, Xiaoyaosan group and fluoxetine group, with 6 rats in each group. Chronic restraint stress was used to prepare rat model of liver-qi stagnation and spleen-deficiency syndrome for 21 days. The expression of TPH2 and IDO1 in hippocampus of rats in each group were detected by real-time PCR and Western-blot. Results: The expression level of TPH2 mRNA in hippocampus of model group was lower than that of the other three groups (P <0.05), and Xiaoyaosan and fluoxetine up-regulated the expression of TPH2 mRNA in model group. IDO1 mRNA (P <0.01), Xiaoyao San and fluoxetine had significant effect on the down-regulation of IDO1 mRNA expression (P <0.01), and Xiaoyao San down-regulated the expression of IDO1 mRNA more significantly. The expression of TPH2 protein in hippocampus of model group, Xiaoyao powder group and fluoxetine group was lower than that in normal group (P <0.01), while TPH2 protein in Xiaoyao powder group was higher than that in model group (P <0.05) IDO1 protein expression was significantly higher than the other three groups (P <0.01). Conclusion: The expression of TPH2 in the hippocampus of rats with liver-qi and spleen-deficiency syndrome is decreased and the expression of IDO1 is increased. Xiaoyaosan may play a therapeutic role by regulating the expression levels of TPH2 and IDO1 in the hippocampus and thus the content of 5-HT.