卒中后抑郁大鼠小脑浦肯野细胞形态学改变及神经元凋亡情况研究

Morphological Changes of Cerebellar Purkinje Cells and Neuronal Apoptosis in Rats with Post- stroke Depression

目的观察卒中后抑郁(PSD)大鼠小脑浦肯野细胞的形态学改变,并检测神经元的凋亡情况。方法选用健康雄性SD大鼠72只,随机分为假手术组(给予假手术处理)、卒中组(行大脑中动脉闭塞术)、抑郁组〔给予慢性不可预见的温和刺激(CUMS)结合孤养〕和PSD组(大脑中动脉闭塞术后给予CUMS结合孤养)。应激(CUMS)后第1、7、14、21天测量体质量,并行蔗糖水实验和旷野实验(OFT)观察大鼠的行为学变化。造模成功后行尼氏染色观察小脑浦肯野细胞的形态学改变,TUNEL染色检测大鼠小脑浦肯野细胞的凋亡情况,并使用免疫组织化学染色法检测各组大鼠浦肯野细胞激活型半胱氨酸天冬氨酸蛋白酶3(activated-caspase 3)的表达。结果与假手术组比较,卒中组、抑郁组及PSD组大鼠应激后第21天体质量、糖水消耗比例、水平运动及垂直运动均降低(P<0.05);与卒中组比较,抑郁组、PSD组大鼠应激后第21天上述指标均降低(P<0.05)。与假手术组比较,卒中组、抑郁组、PSD组均有尼氏体减少甚至消失。卒中组、抑郁组、PSD组浦肯野细胞activated-caspase 3阳性细胞数及凋亡率均高于假手术组,抑郁组、PSD组又高于卒中组(P<0.05)。结论 PSD大鼠浦肯野细胞存在明显的凋亡损伤,该损伤可能与PSD发病相关。

Objective To observe the morphological changes of cerebellar Purkinje cells,and to detect the apoptotic neurons in rats with post- stroke depression（PSD）.Methods A total of 72 healthy male Sprague- Dawley（SD） rats were randomly divided into four groups：sham operated group（sham operated model were given the sham operation）,stroke group（stroke model were set up by blocking the middle cerebral artery）,depression group 〔depression model were raised separately and were given chronic unpredictable mild stress（CUMS） 〕,and PSD group（the middle cerebral artery were blocked,then PSD model were raised separately and were given chronic unpredictable mild stress）.On the 1st,7th,14th and 21st day after CUMS,body weight was measured,to observe the behavioral changes of rats,open- field test（OFT） and the sucrose consumption test were employed.After the completion of model,morphologcal changes of cerebellar Purkinje cells were detected by Nissl staining,TUNEL staining were used to explore the apoptotic changes in Purkinje cells,expression of activated- caspase 3 in Purkinje cells were detected by immunohistochemical staining.Results On the 21st day after CUMS,the body weight,sugar consumption rate and the scores of horizontal movement and vertical movement in stroke group,depression group and the PSD group were significantly lower than those in sham operated group（P〈0.05）;On the 21st day after CUMS,the above indicators in depression group and PSD group were significantly lower than those in stroke group（P〈0.05）.Compared with the sham operated group,the number of Nissl bodies reduced or disappeared in stroke group,depression group and the PSD group.The number of activated- caspase 3 positive Purkinje cells and apoptosis rate of Purkinje cells in stroke group,depression group and PSD group were significantly higher than those in sham operated group,meanwhile,the above indicators in depression group and PSD group were significantly higher than those in stroke group（P〈0.05）.Conclusion Purkinje cells in rats with PSD have obvious apoptotic damage,the damage maybe associated with the onset of PSD.