神经复元方对H_(2)O_(2)诱导的大鼠海马神经元损伤后雄激素受体活化的影响

Study on the effects of the Shenjing Fuyuan Decoction on the androgen receptor in H_(2)O_(2)-induced hippocampal neuron injury in rats

[目的]探究神经复元方对过氧化氢(H_(2)O_(2))诱导的SD大鼠海马神经元损伤中雄激素受体(AR)的活化影响,探讨其治疗卒中后抑郁的作用机制。[方法]取新生24 h的SD大鼠的海马组织,分离原代大鼠海马神经元,通过微管相关蛋白2(MAP-2)免疫荧光鉴定细胞。对SD大鼠给予神经复元方灌胃处理,分离血清,获得空白血清和含药血清,再利用细胞计数试剂(CCK8)检测确定含药血清的最佳处理浓度。使用100μmol/L的H_(2)O_(2)对大鼠海马神经元细胞处理致损伤,分为空白血清组、损伤模型+空白血清组、损伤模型+含药血清组、损伤模型+氟西汀4组。利用蛋白质免疫印迹法(Western blot)和免疫荧光检测各组AR蛋白磷酸化及非磷酸化的情况。[结果]与空白血清组相比,损伤模型+空白血清组的AR、p-AR蛋白表达显著下降。与损伤模型+空白血清组相比,损伤模型+含药血清组和损伤模型+氟西汀组的AR、p-AR蛋白表达上升。[结论]神经复元方对H_(2)O_(2)诱导的SD大鼠海马神经元损伤有一定修复作用,能够促进AR活化,提示AR可能为神经复元方治疗脑卒中后抑郁的重要靶蛋白。

[Objective]To explore the activation effects of the neurorestorative formula on androgen receptor(AR)in H_(2)O_(2)-induced hippocampal neuron injury in Sprague-Dawley(SD)rats and to investigate its mechanism of action in the treatment of post-stroke depression(PSD).[Methods]The hippocampal tissues of 24-hour-old SD rats were taken and primary rat hippocampal neurons were isolated and identified by MAP-2 immunofluorescence.Shenjing Fuyuan Decoction was orally administered to SD rats,and serum was separated to obtain blank serum and drug serum.CCK8 was used to determine the optimal treatment concentration of blank serum and drug serum.A concentration of 100μmol/L H_(2)O_(2) was used to induce damage to rat hippocampal neurons.Western blot and immunofluorescence were used to determine the changes in phosphorylated and non-phosphorylated AR proteins in the blank serum group and H_(2)O_(2)+blank serum/drug serum/fluoxetine group.[Results]Compared with the blank serum group,the expression of AR and p-AR proteins in the H_(2)O_(2)+blank serum group was significantly decreased.Compared with the H_(2)O_(2)+blank serum group,the expression of AR and p-AR proteins in the H_(2)O_(2)+drug serum group and H_(2)O_(2)+fluoxetne group increased.[Conclusion]Shenjing Fuyuan Decoction has a certain repairing effect on H_(2)O_(2)-induced hippocampal neuron injury in SD rats,and it can promote AR activation,indicating that AR may be an important target protein for the treatment of PSD with the Shenjingfuyuan Decoction.