醒脾解郁方对抑郁大鼠免疫功能的影响及机制研究

The rapeutic effect and mechanism of Xingpi Jieyu formula on immune function of depression model rats

目的研究醒脾解郁方对抑郁模型大鼠抑郁症行为学指征以及免疫功能的影响,并探索其可能的作用机制。方法将40只SPF级SD雄性大鼠随机分为空白对照组、模型组、西酞普兰组、醒脾解郁方组,每组10只。空白对照组不予刺激,其余3组应用慢性不可预见轻度应激(chronic unpredictable mild stress,CUMS)的方法制备抑郁模型。造模同时灌胃给药,西酞普兰组和醒脾解郁方组分别以西酞普兰和醒脾解郁方灌胃治疗,空白对照组、模型组给予等剂量0.9%氯化钠溶液灌胃。采用糖水偏好实验、旷场实验、强迫游泳实验评价大鼠行为学的变化,苏木精—伊红染色观察脾组织形态变化,免疫组化检测脾脏Foxp3的表达情况,流式细胞术检测脾脏、外周血中CD_(3)^(+)T细胞、CD_(4)^(+)T细胞以及CD_(8)^(+)T细胞的变化。结果与空白对照组相比,模型组大鼠糖水偏好率、旷场实验直立次数和穿越次数均降低(P<0.05),强迫游泳不动时间升高(P<0.05);脾组织结构中度异常,红髓、白髓界限不清;脾脏Foxp3阳性率升高(P<0.05),脾脏以及外周血CD_(3)^(+)T细胞、CD_(4)^(+)T细胞及脾脏CD_(8)^(+)T细胞比例降低(P<0.05)。与模型组比较,醒脾解郁方组大鼠糖水偏好率、旷场实验直立次数和穿越次数均升高(P<0.05),强迫游泳不动时间降低(P<0.05);脾组织结构异常程度减轻,红髓、白髓分布均匀,界限较清楚;脾脏Foxp3阳性率降低(P<0.05),脾脏及外周血CD_(3)^(+)T细胞、CD_(4)^(+)T细胞比例升高(P<0.05)。结论醒脾解郁方能够改善抑郁模型大鼠抑郁样行为,其机制可能与改变T细胞亚群的比例,改善免疫功能有关。

Objective To investigate the therapeutic effect of behavioral indications of depression and immune function of depression model rats and its possible mechanism of Xingpi Jieyu formula.Methods Forty SPF-grade SD male rats were randomly divided into blank control group,model group,citalopram group,and Xingpi Jieyu formula group,with ten rats in each group.The blank control group was not stimulated,while the remaining three groups were given chronic unpredictable mild stress(CUMS)to establish depression model rats.Drugs was administered through Gastric gavage at the same time as molding.The Citalopram group and the Xingpi Jieyu formula group were given citalopram and Xingpi Jieyu formula respectively,while the blank control group and the model group were given the same dose of 0.9%sodium chloride solution.The behavioral changes of rats were evaluated by sugar water preference experiment,open field experiment and forced swimming experiment,spleen tissue morphological changes were observed by the method HE staining,the expression of Foxp3 in spleen was detected by immunohistochemistry,and the changes of CD_(3)^(+)T cells,CD_(4)^(+)T cells and CD_(8)^(+)T cells in spleen and peripheral blood were detected by flow cytometry.Results Compared with the blank control group,the rate of sugar water preference,times of upright and crossing in open field experiments were decresed(P<0.05),and the immobile time of forced swimming was increased(P<0.05),the spleen tissue structure was moderately abnormal,the boundaries between red and white pulp were not clear,the positive rate of Foxp3 in the spleen was increased(P<0.05),the proportion of CD_(3)^(+)T cells and CD_(4)^(+)T cells in the spleen and peripheral blood was decresed(P<0.05),and proportion of CD_(8)^(+)in the spleen was decresed(P<0.05)in the model group.Compared with the model group,the sugar water preference rate,times of upright crossings in open field experiment were increased(P<0.05),and the time of forced swimming was decresed(P<0.05),the degree of spleen tissue structure abnormality was decresed,red pulp and white pulp were evenly distributed,the boundaries were clearer,the positive rate of Foxp3 in the spleen was decresed(P<0.05),and the proportion of CD_(3)^(+)T cells and CD_(4)^(+)T cells in the spleen and peripheral blood was increased(P<0.05)in the Xingpi Jieyu formula group.Conclusion Xingpi Jieyu formula can improve depressive behaviors of depression model rats,and the mechanism may be related to changing the proportion of T cell subsets to improve the immune function.