Objective: Chaihu Shugan San (CSS) has a long history for treating major depressive disorder (MDD), which has been verified effectively and safely in clinical studies. Deficient angiogenesis plays important roles in MDD. However, the underlying mechanisms of CSS on angiogenesis remain poorly understood.
Methods: Network pharmacology analysis was applied to explore the potential angiogenic targets and pathways between CSS and MDD. These targets would be validated in chronic unpredictable mild stress (CUMS)-induced depressive-like mice by Western blots, immunofluorescence, and immunohistochemistry. Then, the underlying molecular mechanisms were further investigated in brain microvascular endothelial cells (BMVECs) with CSS-containing serum by Western blots and immunofluorescence.
Results: Network pharmacology analysis showed that the antidepressant role of CSS was closely associated with Silent information regulator protein 1 (SIRT1)/Forkhead box O1 (FOXO1) axis-mediated angiogenesis. This prediction was confirmed in the following experiments. CSS induced angiogenesis, increased SIRT1 expression, and decreased FOXO1 expression in the hippocampus of CUMS mice. Five percent CSS-containing serum produced a significant increase in BMVECs proliferation, migration, and tube formation, but these effects were reduced by SIRT1 silencing. CSS serum could also promote FOXO1 translocation to the cytoplasm through SIRT1 signaling, which triggered FOXO1 protein degradation. What is more, CSS upregulated VEGFA and BDNF expressions not only in the hippocampus of depressive mice but also in BMVECs supernatants. Of note, these trophic factors play important roles in promoting neurogenesis.
Conclusion: The study showed that CSS could promote angiogenesis and neurogenesis in the hippocampus of CUMS-induced mice. The underlying molecular mechanism involves the SIRT1/FOXO1 axis and subsequent regulation of VEGFA and BDNF. These findings provide novel insight into CSS drug development, and targeting the SIRT1/FOXO1 axis might be a promising strategy to treat MDD.
Graphical Abstract:

Keywords: major depressive disorder, Chaihu Shugan San, SIRT1/FOXO1 axis, angiogenesis, brain microvascular endothelial cell


中文翻译：

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SIRT1/FOXO1 轴介导的海马血管生成参与柴胡疏肝散的抗抑郁作用

目的：柴胡舒肝散（CSS）治疗重度抑郁症（MDD）的历史悠久，已在临床研究中得到有效、安全的验证。血管生成缺陷在 MDD 中起重要作用。然而，CSS 对血管生成的潜在机制仍然知之甚少。
方法：应用网络药理学分析探讨CSS和MDD之间潜在的血管生成靶点和通路。这些目标将通过蛋白质印迹、免疫荧光和免疫组织化学在慢性不可预测的轻度应激 (CUMS) 诱导的抑郁样小鼠中得到验证。然后，通过蛋白质印迹和免疫荧光进一步研究了含有 CSS 血清的脑微血管内皮细胞 (BMVEC) 的潜在分子机制。
结果：网络药理学分析表明，CSS的抗抑郁作用与沉默信息调节蛋白1（SIRT1）/叉头盒O1（FOXO1）轴介导的血管生成密切相关。这一预测在以下实验中得到证实。CSS 在 CUMS 小鼠的海马中诱导血管生成、增加 SIRT1 表达并降低 FOXO1 表达。5% 的含 CSS 的血清显着增加了 BMVEC 的增殖、迁移和管形成，但这些影响因 SIRT1 沉默而降低。CSS血清还可以通过SIRT1信号促进FOXO1易位至细胞质，从而引发FOXO1蛋白降解。更重要的是，CSS不仅在抑郁小鼠的海马中上调了VEGFA和BDNF的表达，而且在BMVECs上清液中也有上调。值得注意的是，
结论：研究表明，CSS可以促进CUMS诱导小鼠海马的血管生成和神经发生。潜在的分子机制涉及 SIRT1/FOXO1 轴以及随后对 VEGFA 和 BDNF 的调节。这些发现为 CSS 药物开发提供了新的见解，靶向 SIRT1/FOXO1 轴可能是治疗 MDD 的有希望的策略。
图解摘要：

关键词：重度抑郁症，柴胡舒肝散，SIRT1/FOXO1轴，血管生成，脑微血管内皮细胞