目的：本文主要探讨白芍提取物对PMS肝气郁证大鼠下丘脑Cav1.2钙通道关键蛋白CACNA1C及其下游信号通路中CaM、BDNF蛋白表达和对CaMK Ⅱ磷酸化的影响，从而明确白芍提取物治疗肝气郁结证的分子作用靶点。方法：利用慢性束缚应激法制备PMS肝气郁证大鼠模型，使用白芍提取物进行药物干预。模型制备成功后检测下丘脑CACNA1C蛋白表达，以及下游信号通路中CaM、BDNF蛋白表达和CaMK Ⅱ的磷酸化水平。离体培养海马原代神经元，通过KCl激活L型钙通道，检测白芍提取物中的有效成分单体芍药苷对细胞内钙超载的影响。结果：PMS肝气郁证大鼠下丘脑CACNA1C蛋白表达增加，CaMK Ⅱ磷酸化水平提高，BDNF表达减少，说明白芍提取物可显著改善上述蛋白表达异常的现象，抑制KCl激活的L型钙通道引起的细胞内钙离子浓度的增加。结论：白芍提取物可能是通过调控细胞内Cav1.2，抑制其下游CaM/CaMK Ⅱ信号通路的活化发挥治疗PMS肝气郁证的作用。

This study aimed to explore the impacts of the extracts from Radix Paeoniae Alba on hypothalamic CACNA1C,the key protein of Cav1.2 channel,and hypothalamic CaM and BDNF in its downstream,as well as the phosphorylation of CaMK Ⅱ in the hypothalamus in PMS rats with liver-qi depression.On this basis,the molecular targets of the extracts engaged in the treatment was confirmed.The PMS model rats with liver-qi depression were caused by chronic restraint stress taking the extracts from Radix Paeoniae Alba as its treatment.When verified success of the modeling of liver-qi depression,the hypothalamic protein expressions of CACNA1C,CaM and BDNF of its downstream and the phosphorylation of CaMK Ⅱ were detected.Then the hippocampal primary neurons were cultivated in vitro,in which the L-type calcium channels was activated by KCl to test the effects of paeoniflorin monomer,one of the active ingredients in the extracts,on intercellular calcium overload in the hippocampal neurons.In the experiment,it was found that the protein expressions of hypothalamus CACNA1C and p-CaMK II increased,while hypothalamus BDNF decreased significantly,which indicated that the extract of Radix Paeoniae Alba could regulate the abnormal expressions of the aforementioned proteins and inhibit the augmented intercellular calcium concentration caused by the activation the L-type calcium channels stimulated by KCl.In conclusion,it was demonstrated that the extract of Radix Paeoniae Alba could suppress the activation of CaM/CaMK Ⅱ signaling in its downstream by the regulation of intercellular Cav1.2 channels to achieve the efficacy of mitigating PMS liver-qi depression in rats.

国家自然科学基金委青年科学基金项目（81473359）：Cav1.2介导的CaM/CaMK Ⅱ钙信号通路在PMS肝气郁证中的作用及舒郁胶囊干预机制研究，负责人：宋春红；山东省卫生厅中医药科技发展计划项目任务书（2013-025）：舒郁胶囊对抑郁症模型大鼠海马脑区钙调蛋白依赖的蛋白激酶Ⅱ（CaMKII）表达和分布的影响，负责人：宋春红。