基于海马CA1区微量注射AMPA对肝郁脾虚证模型大鼠杏仁核BLA区阳性细胞的影响探讨逍遥散的调节机制  

Regulation mechanism of Xiaoyao San studied based on influence of microinjection of AMPA in hippocampus CA1 zone on positive cells in amygdaloid nucleus BLA zone in rat model with syndrome of liver depression and spleen deficiency

作　　者：岳利峰[1] 王玲[2] 奚胜艳[3] 王福[1] 陈昌乐[1] 王琦 赵健[1] 

机构地区：[1]北京中医药大学,北京100029 [2]北京航空航天大学 [3]厦门大学 [4]山西省实验中学

出　　处：《北京中医药大学学报》2012年第11期743-746,751,I0002,共6页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家自然科学青年基金项目(No.61101008);北京中医药大学自主选题项目(No.2009JYBZZ-JS006)

摘　　要：目的运用免疫组化方法,对比海马CA1区微量注射α-氨基羟甲基恶唑丙酸(AMPA)对肝郁脾虚证模型大鼠杏仁核BLA区AMPA受体亚基阳性细胞的影响,探讨逍遥散调节肝郁脾虚证的机制。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、AMPA组和逍遥散组。以21 d慢性束缚应激方法造肝郁脾虚证模型,于模型大鼠双侧海马CA1区,埋管微量注射AMPA受体的激动剂AMPA,逍遥散组造模方法和AMPA组尽可能相似,突出逍遥散和AMPA干预的可比性。运用免疫组化方法检测杏仁核基底外侧区(BLA区)AMPA受体的2个重要亚基谷氨酸受体1(GluR1)和谷氨酸受体2(GluR2)的阳性细胞数变化情况,比较逍遥散组和AMPA组上述指标的变化趋势是否一致。结果造模21 d后,在杏仁核BLA区,与正常组比较,AMPA组和逍遥散组GluR1和GluR2阳性细胞数表达差异均无统计学意义(均P>0.05);与模型组比较,AMPA组和逍遥散组GluR1和GluR2阳性细胞数表达差异均有统计学意义(均P<0.01),且2组GluR1阳性细胞数表达均较模型组增高,GluR2阳性细胞数表达均较模型组降低,2组阳性细胞数表达升降趋势一致。各组阳性细胞积分光密度结果与阳性细胞数结果一致。结论结合前期实验,从免疫组化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,从而治疗肝郁脾虚证。Objective To compare the influence of microinjection of AMPA in hippocampus CA1 zone on subunit positive cells of AMPA in amygdaloid nucleus BLA zone in rat model with syndrome of liver depression and spleen deficiency by using immunohistochemistry assay, and investigate the mechanism of Xiaoyao San in regulation of syndrome of liver depression and spleen deficiency. Methods Male SD rats （n = 25） were randomly divided into normal group, model group, sham-operation group, AMPA group and Xiaoyao San group. The model of syndrome of liver depression and spleen deficiency was established with 21-day chronic constraint stress in model group and microinjection of AMPA in CA1 zone of bilateral hippocampus. The modeling method in Xiaoyao San group was as the same as possible with that in AMPA group for highlighting the comparability between Xiaoyao San and AMPA. The changes of positive cell number of GluR1 and GluR2 in amygdaloid BLA zone were detected by using immunohistochemistry technique, and whether the change trend of above indexes was consistent was compared between AMPA group and Xiaoyao San group. Results After modeling for 21 days, the differences in the expressions of positive cells of GluR1 and GluR2 had no statistical significance between AMPA group or Xiaoyao San group and normal group （all P 〉0.05 ）. The differences in the expressions of positive cells of GluR1 and GluR2 had statistical significance between AMPA group or Xiaoyao San group and model group （all P 〈0.01）. The expression of positive cell of GluR1 was higher, and expression of positive cell of GluR2 was lower in AMPA group and Xiaoyao San group than those in model group, so the increasing and decreasing trends of positive cell expressions were consistent in these two groups. The results of integral optical density of positive cells were consistent with the results of positive cell counts in all groups. Conclusion Based on early research and from the angle of immunohistochemistry, it can be deduced further that Xiaoyao San can tr

关 键 词：逍遥散 肝郁脾虚证 调节机制 免疫组化 GLUR1 GLUR2 大鼠 

分 类 号：R285.5[医药卫生—中药学]