Objective To investigate the effect of Xingnao Jieyu Decoction on the function of Notch/Hes signaling pathway in hippocampus of rats with post-stroke depression (PSD).Methods A total of 80 healthy male specific pathogen-free Wistar rats were randomly divided into control group,model group,prozac group and Xingnao Jieyu Decoction group;20 rats in each group.The rats in the model group,prozac group and Xingnao Jieyu Decoction group were used to prepare the cerebral ischemia model by middle cerebral artery occlusion method,and then the rats were used to prepare the depressive model by a variety of chronic unpredictable stimuli.The rats in the control group did not have any intervention.After the preparation of PSD model,the rats in the control group and model group were given saline 2.5 mL by intragastric administration,the rats in prozac group were given prozac 2.5 mL (1.8 mg · kg-1) by intragastric administration,the rats in Xingnao Jieyu Decoction group were given Xingnao Jieyu Decoction 2.5 mL (150.0 mg · kg-1) by intragastric administration;one times daily for 28 days.Ten rats in each group were sacrificed immediately on the nineteenth and twenty-eighth day of drug intervention,and the hippocampus tissues were isolated.The expression of Notch and Hes mRNA and protein in hippocampus were detected by real-time quantitative polymerase chain reaction and Western blot method.The open probability,current amplitude,opening time and closing time of Notch signaling pathway were detected by patch clamp technique.Results On the nineteenth and twenty-eighth day of drug intervention,the expression of Notch and Hes protein and mRNA in hippocampus of rats in the model group,prozac group and Xingnao Jieyu Decoction group was significantly higher than that in the control group (P ＜ 0.05);the expression of Notch and Hes protein and mRNA in hippocampus of rats in the prozac group and Xingnao Jieyu Decoction group was significantly higher than that in the model group (P ＜ 0.05);the expressions of Notch and Hes protein and mRNA in hippocampus of rats in the Xingnao Jieyu Decoction group was significantly higher than that in the prozac group (P ＜ 0.05).There was no significant difference in Notch and Hes protein and mRNA expression in hippocampus of rats between the nineteenth and twenty-eighth day of drug intervention (P ＞ 0.05).The expression of Notch and Hes protein and mRNA in hippocampus of rats on the twenty-eighth day was significantly higher than that on the nineteenth day in the model group,prozac group and Xingnao Jieyu Decoction group (P ＜ 0.05).Compared with the control group,the open probability and current amplitude of Notch signaling pathway in hippocampus of rats increased significantly,the opening time prolonged significantly,and the closing time shortened significantly in the model group,prozac group and Xingnao Jieyu Decoction group on the nineteenth and twenty-eighth day of drug intervention (P ＜ 0.05).Compared with the model group,the open probability and current amplitude of Notch signaling pathway in hippocampus of rats increased significantly,the opening time prolonged significantly,and the closing time shortened significantly in the prozac group and Xingnao Jieyu Decoction group on the nineteenth and twenty-eighth day of drug intervention (P ＜ 0.05).Compared with the prozac group,the open probability and current amplitude of Notch signaling pathway in hippocampus of rats increased significantly,the opening time prolonged significantly,and the closing time shortened significantly in the Xingnao Jieyu Decoction group on the nineteenth and twenty-eighth day of drug intervention (P ＜ 0.05).There was no significant difference in the open probability,current amplitude,opening time and closing time of Notch signaling pathway between the nineteenth and twenty-eighth day of drug intervention in the control group(P ＞ 0.05).Compared with the nineteenth day of drug intervention,the open probability and current amplitude of Notch signaling pathway in hippocampus of rats increased significantly,the opening time prolonged significantly,and the closing time shortened significantly on the twenty-eighth day of drug intervention in the model group,prozac group and Xingnao Jieyu Decoction group (P ＜ 0.05).Conclusion Xingnao Jieyu Decoction can mediate the activation of Notch / Hes signaling pathway in hippocanpus of PSD rats,thus effectively promote the recovery of neurological function.Notch/Hes signaling pathway is expected to be an important target for drug action.%目的 探讨醒脑解郁方对卒中后抑郁(PSD)大鼠海马组织中Notch/Hes信号通路功能的影响.方法 将80只健康雄性无特定病原体级Wistar大鼠随机分为对照组、模型组、百忧解组和醒脑解郁方组,每组20只.模型组、百忧解组和醒脑解郁方组大鼠采用大脑中动脉线栓法制备大鼠脑缺血模型,再给予多种慢性不可预见性刺激制备抑郁症模型;对照组大鼠不予任何干预.PSD模型制备完成后,对照组和模型组大鼠给予生理盐水2.5 mL灌胃,百忧解组大鼠给予百忧解1.8 mg·kg-1溶于2.5 mL生理盐水中灌胃,醒脑解郁方组大鼠给予醒脑解郁方150.0 mg·kg-1溶于2.5 mL生理盐水中灌胃,均为每日1次,连续灌胃28 d.药物干预第19、28天,每组各选择10只大鼠立即处死,分离海马组织,采用实时荧光定量聚合酶链反应法和Western blot法检测海马组织中Notch和Hes mR-NA及蛋白表达水平,采用膜片钳技术检测Notch信号通路开放概率、电流幅度、开放时间和关闭时间.结果 药物干预第19、28天,模型组、百忧解组和醒脑解郁方组大鼠海马组织中Notch和Hes蛋白及mRNA表达显著高于对照组(P＜0.05),百忧解组和醒脑解郁方组大鼠海马组织中Notch和Hes蛋白及mRNA表达显著高于模型组(P＜0.05),醒脑解郁方组大鼠海马组织中Notch和Hes蛋白及mRNA表达显著高于百忧解组(P＜0.05).对照组大鼠第28天海马组织中Notch和Hes蛋白及mRNA表达与第19天比较差异均无统计学意义(P＞0.05);模型组、百忧解组和醒脑解郁方组大鼠第28天海马组织中Notch和Hes蛋白及mRNA表达显著高于第19天(P＜0.05).药物干预第19、28天,模型组、百忧解组和醒脑解郁方组大鼠海马组织中Notch信号通路开放概率、电流幅度显著高于对照组,开放时间显著长于对照组,关闭时间显著短于对照组(P＜0.05);百忧解组和醒脑解郁方组大鼠海马组织中Notch信号通路开放概率、电流幅度显著高于模型组,开放时间显著长于模型组,关闭时间显著短于模型组(P＜0.05);醒脑解郁方组大鼠海马组织中Notch信号通路开放概率、电流幅度显著高于百忧解组,开放时间显著长于百忧解组,关闭时间显著短于百忧解组(P＜0.05).对照组大鼠第19、28天海马组织中Notch信号通路开放概率、电流幅度、开放时间及关闭时间比较差异均无统计学意义(P＞0.05);模型组、百忧解组和醒脑解郁方组大鼠第28天海马组织中Notch信号通路开放概率、电流幅度显著高于第19天,开放时间显著长于第19天,关闭时间显著短于第19天(P＜0.05).结论 醒脑解郁方能够介导PSD大鼠海马组织中Notch/Hes信号通路活化,从而有效促进神经功能恢复.Notch/Hes信号通路有望成为药物作用的重要靶点.
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