加味四逆散抗应激性抑郁效应及其海马NMDA受体通道机制的初步研究

目的研究加味四逆散(JWSNS)抗应激性抑郁效应及其海马N-甲基-D-天冬氨酸(N-methyl-D-aspartate,NMDA)受体通道的机制。方法采用慢性轻度不可预计应激(chronicmild unpredictable stress,CMUS)制作大鼠抑郁症模型。在造模前后测量基础糖水消耗量和体重的变化;离体海马脑片TTC染色检测各组大鼠海马损伤情况以及JWSNS含药血清的保护作用;运用细胞贴附式膜片钳记录模式检测大鼠海马NMDA受体通道开放概率和平均开放时间的变化,Tillvision图像处理系统检测海马神经元细胞内游离Ca2+浓度。结果CMUS可引起大鼠糖水偏爱度下降(P<0.01),JWSNS和MK801可提高应激性抑郁症大鼠糖水偏爱度(P<0.01,P<0.05),但对大鼠的体重无影响。JWSNS、20%JWSNS含药血清和MK801均能明显升高应激性抑郁症大鼠海马光密度值,降低海马脑片损伤百分率(P<0.05,P<0.01)。模型组大鼠海马NMDA受体通道开放概率明显增高(P<0.05),而JWSNS、MK801能明显降低海马NMDA受体通道开放概率(P<0.05,P<0.01);对NMDA受体通道平均开放时间无明显影响。JWSNS、MK801能明显降低应激性抑郁症大鼠海马神经元内升高的游离Ca2+浓度(P<0.01)。结论JWSNS具有抗抑郁作用,NMDA受体可能是其药理学作用靶点之一,调控海马神经元NMDA受体的功能状态可能是JWSNS发挥抗抑郁效应的直接作用机制之一。

The initial research on the effect of anti-stressed depression of Jiaweisinisan and it's mechanism of N-methyl-D-aspartate receptor channel in hippocampus

Aim To study the anti-stress depression effect of Jiaweisinisan(JWSNS)and it's mechanism of N-methyl-D-aspartate(NMDA)receptor channel in hippocampus.Methods Chronic mild unpredictable stress(CMUS)was adopted to establish the rat depres-sion model.Before and after model establishing,the sucrose consumptions and the weight of rats were detected.TTC dyeing of hippocampus brain slices ex vivo were used to detect the situation of hippocampus neuron damage and the protective effect of serum containing components of JWSNS.The opening probability and the average opening time of NMDA receptor channels were detected by attach on cell patch clamp,the Ca2+ concentrations in single cell of hippocampus neurons were detected by Tillvis ION software.Results CMUS could decrease the sucrose preference and the weights of rats(P<0.01),JWSNS and MK801 could increase sucrose preference,but had no effects on the weights.The formula of JWSNS,the 20% serum containing components of JWSNS and MK801 could significantly increase the OD values of hippocampus brain slices TTC staining and decrease the hippocampus neuron damage rates(P<0.05,P<0.01).The opening probability of NMDA receptor channels in model group was higher than that of normal group(P<0.05),while JWSNS and MK801 could significantly decrease the opening probability of NMDA receptor channels in hippocampus(P<0.05,P<0.01).But there were no statistical significance among groups in the average opening time of NMDA receptor channels.Meanwhile,JWSNS and MK801 could obviously decrease the Ca2+ concentration in single cell of hippocampus neurons(P<0.01).Conclusion JWSNS has the effect of anti-stress depression.It is suggested that NMDA receptor may be one of the pharmacological targets of JWSNS for anti-stress depression by directly regulating the function of NMDA receptor.