Abstract：
OBJECTIVE: Ganoderic Acid A (GAA), a primary bioactive component in Ganoderma, has demonstrated ameliorative effects on depressive-like behaviors in a Chronic Social Defeat Stress (CSDS) mouse model. This study aims to elucidate the underlying molecular mechanisms through proteomic analysis.
METHODS: C57BL/6 J mice were allocated into control (CON), chronic social defeat stress (CSDS), GAA, and imipramine (IMI) groups. Post-depression induction via CSDS, the GAA and IMI groups received respective treatments of GAA (2.5 mg/kg) and imipramine (10 mg/kg) for five days. Behavioral assessments utilized standardized tests. Proteins from the prefrontal cortex were analyzed using LC-MS, with further examination via bioinformatics and PRM for differential expression. Western blot analysis confirmed protein expression levels.
RESULTS: Chronic social defeat stress (CSDS) induced depressive-like behaviors in mice, which were significantly alleviated by GAA treatment, comparably to imipramine (IMI). Proteomic analysis identified distinct proteins in control (305), GAA-treated (949), and IMI-treated (289) groups. Enrichment in mitochondrial and synaptic proteins was evident from GO and PPI analyses. PRM analysis revealed significant expression changes in proteins crucial for mitochondrial and synaptic functions (namely, Naa30, Bnip1, Tubgcp4, Atxn3, Carmil1, Nup37, Apoh, Mrpl42, Tprkb, Acbd5, Dcx, Erbb4, Ppp1r2, Fam3c, Rnf112, and Cep41). Western blot validation in the prefrontal cortex showed increased levels of Mrpl42, Dcx, Fam3c, Ppp1r2, Rnf112, and Naa30 following GAA treatment.
CONCLUSIONS: GAA exhibits potential antidepressant properties, with its action potentially tied to the modulation of synaptic functions and mitochondrial activities.
METHODS: C57BL/6 J mice were allocated into control (CON), chronic social defeat stress (CSDS), GAA, and imipramine (IMI) groups. Post-depression induction via CSDS, the GAA and IMI groups received respective treatments of GAA (2.5 mg/kg) and imipramine (10 mg/kg) for five days. Behavioral assessments utilized standardized tests. Proteins from the prefrontal cortex were analyzed using LC-MS, with further examination via bioinformatics and PRM for differential expression. Western blot analysis confirmed protein expression levels.
RESULTS: Chronic social defeat stress (CSDS) induced depressive-like behaviors in mice, which were significantly alleviated by GAA treatment, comparably to imipramine (IMI). Proteomic analysis identified distinct proteins in control (305), GAA-treated (949), and IMI-treated (289) groups. Enrichment in mitochondrial and synaptic proteins was evident from GO and PPI analyses. PRM analysis revealed significant expression changes in proteins crucial for mitochondrial and synaptic functions (namely, Naa30, Bnip1, Tubgcp4, Atxn3, Carmil1, Nup37, Apoh, Mrpl42, Tprkb, Acbd5, Dcx, Erbb4, Ppp1r2, Fam3c, Rnf112, and Cep41). Western blot validation in the prefrontal cortex showed increased levels of Mrpl42, Dcx, Fam3c, Ppp1r2, Rnf112, and Naa30 following GAA treatment.
CONCLUSIONS: GAA exhibits potential antidepressant properties, with its action potentially tied to the modulation of synaptic functions and mitochondrial activities.
摘要：
目的：灵芝酸A（GAA），灵芝的主要生物活性成分,在慢性社会失败应激（CSDS）小鼠模型中已证明对抑郁样行为的改善作用。本研究旨在通过蛋白质组学分析阐明潜在的分子机制。
方法：将C57BL/6J小鼠分配到对照（CON）中，慢性社会失败压力(CSDS)，GAA,和丙咪嗪（IMI）组。通过CSDS诱导抑郁后，GAA和IMI组分别接受GAA(2.5mg/kg)和丙咪嗪(10mg/kg)治疗5天.行为评估采用标准化测试。使用LC-MS分析来自前额叶皮质的蛋白质,通过生物信息学和PRM进一步检查差异表达。蛋白质印迹分析证实了蛋白质表达水平。
结果：慢性社会失败应激（CSDS）诱导小鼠抑郁样行为，GAA治疗可显着缓解，与丙咪嗪（IMI）相比。蛋白质组学分析鉴定了对照中的不同蛋白质(305)，GAA处理（949），和IMI治疗组（289）。从GO和PPI分析中可以明显看出线粒体和突触蛋白的富集。PRM分析揭示了对线粒体和突触功能至关重要的蛋白质的显着表达变化（即，Naa30,Bnip1,Tubgcp4,Atxn3,Carmil1,Nup37,Apoh,Mrpl42,Tprkb,Acbd5,Dcx,Erbb4,Ppp1r2,Fam3c,Rnf112和Cep41）。蛋白质印迹验证在前额叶皮层显示Mrpl42，Dcx，Fam3c,GAA处理后的Ppp1r2、Rnf112和Naa30。
结论：GAA具有潜在的抗抑郁特性，其作用可能与突触功能和线粒体活动的调节有关。
方法：将C57BL/6J小鼠分配到对照（CON）中，慢性社会失败压力(CSDS)，GAA,和丙咪嗪（IMI）组。通过CSDS诱导抑郁后，GAA和IMI组分别接受GAA(2.5mg/kg)和丙咪嗪(10mg/kg)治疗5天.行为评估采用标准化测试。使用LC-MS分析来自前额叶皮质的蛋白质,通过生物信息学和PRM进一步检查差异表达。蛋白质印迹分析证实了蛋白质表达水平。
结果：慢性社会失败应激（CSDS）诱导小鼠抑郁样行为，GAA治疗可显着缓解，与丙咪嗪（IMI）相比。蛋白质组学分析鉴定了对照中的不同蛋白质(305)，GAA处理（949），和IMI治疗组（289）。从GO和PPI分析中可以明显看出线粒体和突触蛋白的富集。PRM分析揭示了对线粒体和突触功能至关重要的蛋白质的显着表达变化（即，Naa30,Bnip1,Tubgcp4,Atxn3,Carmil1,Nup37,Apoh,Mrpl42,Tprkb,Acbd5,Dcx,Erbb4,Ppp1r2,Fam3c,Rnf112和Cep41）。蛋白质印迹验证在前额叶皮层显示Mrpl42，Dcx，Fam3c,GAA处理后的Ppp1r2、Rnf112和Naa30。
结论：GAA具有潜在的抗抑郁特性，其作用可能与突触功能和线粒体活动的调节有关。
