背景:加味丹芝逍遥散(MDXS)是中国治疗抑郁症的有效临床处方,明代被剥夺了丹芝逍遥散。MDSX对新抗抑郁药的开发具有重要意义,但其药理机制却鲜有研究。
目的:通过网络药理学和体内外实验验证,揭示MDXS治疗抑郁症的活性成分和分子机制。
方法:UPLC-Q-TOF-MS/MS用于鉴定MDXS冻干粉中的化学成分,含药血清,和脑脊液(CSF)。基于对CSF中原型组件的分析,主要组成部分,使用网络药理学和分子对接研究了MDXS治疗抑郁症的潜在治疗靶点和可能的药理学机制。建立皮质酮(CORT)诱导的小鼠抑郁模型,探讨MDXS的抗抑郁作用。培养HT22细胞以验证活性组分的神经保护作用和核心靶标。
结果:MDXS冻干粉中含有81种化合物,血清中36种原型成分,并确定了CSF中的13个原型组件,分别。网络药理学分析表明,CSF中的这13种原型成分与抑郁症共有190个共同靶标,主要富集MAPK和PI3K/AKT信号通路。PPI分析表明AKT1和MAPK1(ERK1/2)是核心目标。分子对接显示壬二酸(AA),senkyunolideA(SA),atractylenolideIII(ATIII),与AKT1和MAPK1的结合能最高。动物实验证实MDXS能逆转CORT诱导的抑郁样行为,改善突触可塑性,减轻海马CA3区神经元损伤,并上调p-ERK1/2和p-AKT的蛋白表达。在HT22细胞中,壬二酸,senkyunolideA,并对CORT引起的细胞损伤有明显的保护作用,并上调p-ERK1/2和p-AKT的蛋白水平。
结论:这些结果表明,MDXS可能部分通过壬二酸发挥抗抑郁作用,senkyunolideA,以及靶向ERK1/2和AKT的白曲内酯III。
BACKGROUND: Modified Danzhi Xiaoyao San (MDXS) is an effective clinical prescription for depression in China, which was deprived of Danzhi Xiaoyao San in the Ming Dynasty. MDSX has significant implications for the development of new antidepressants, but its pharmacological mechanism has been rarely studied.
OBJECTIVE: To reveal the active components and molecular mechanism of MDXS in treating depression through network pharmacology and experimental verification in vivo and in vitro.
METHODS: UPLC-Q-TOF-MS/MS was used to identify the chemical components in the MDXS freeze-dried powder, drug-containing serum, and cerebrospinal fluid (CSF). Based on the analysis of prototype components in the CSF, the major constituents, potential therapeutic targets and possible pharmacological mechanisms of MDXS in treating depression were investigated using network pharmacological and molecular docking. Then corticosterone (CORT)-induced mice model of depression was established to investigate the antidepressant effects of MDXS. HT22 cells were cultured to verify the neuroprotective effects and core targets of the active components.
RESULTS: There were 81 compounds in MDXS freeze-dried powder, 36 prototype components in serum, and 13 prototype components in CSF were identified, respectively. Network pharmacology analysis showed that these 13 prototype components in the CSF shared 190 common targets with depression, which were mainly enriched in MAPK and PI3K/AKT signaling pathways. PPI analysis suggested that AKT1 and MAPK1 (ERK1/2) were the core targets. Molecular docking revealed that azelaic acid (AA), senkyunolide A (SA), atractylenolide III (ATIII), and tokinolide B (TB) had the highest binding energy with AKT1 and MAPK1. Animal experiments verified that MDXS could reverse CORT-induced depression-like behaviors, improve synaptic plasticity, alleviate neuronal injury in hippocampal CA3 regions, and up-regulate the protein expression of p-ERK1/2 and p-AKT. In HT22 cells, azelaic acid, senkyunolide A, and atractylenolide III significantly protected the cell injury caused by CORT, and up-regulated the protein levels of p-ERK1/2 and p-AKT.
CONCLUSIONS: These results suggested that MDXS may exert antidepressant effects partially through azelaic acid, senkyunolide A, and atractylenolide III targeting ERK1/2 and AKT.