背景:开心解郁颗粒(KJG),Kai-xin-san和Si-ni-san的改进公式,是一种非常有效的配方,在以前的研究中证明了预防抑郁症的功效。然而,KJG抗抑郁作用对炎症分子的潜在分子机制尚不清楚.本研究旨在通过网络药理学和实验验证来探讨KJG对抑郁症的治疗作用。
方法:我们采用了多方面的方法,结合高效液相色谱(HPLC),网络药理学,和分子对接,解开KJG抗抑郁作用的潜在机制。为了证实我们的发现,我们对小鼠进行了至少两个独立的体内实验,利用慢性不可预测的轻度应激(CUMS)诱导和脂多糖(LPS)诱导的模型。此外,体内实验的结果通过体外实验进行了验证。行为测试被用来评估抑郁样行为,而Nissl染色用于评估海马的形态变化。使用免疫荧光染色联合测定促炎细胞因子和通路相关蛋白的表达,酶联免疫吸附测定(ELISA),和蛋白质印迹(WB)。
结果:我们基于网络的方法表明,人参皂苷Rg1(GRg1)和柴胡皂苷d(Ssd)是KJG的主要成分,它们通过调节TLR4,PI3K,AKT1和FOXO1通过toll样受体靶向,PI3K/AKT,和FoxO途径。在体内,KJG可以减轻抑郁样行为,保护海马神经元细胞,并减少促炎介质(TNF-α,IL-6和IL-1β)通过抑制TLR4表达,通过核出口抑制FOXO1。此外,KJG增加PI3K的表达水平,AKT,p-PI3K,p-AKT,P-PTEN我们的体外测定与我们的体内研究一致。另一方面,通过应用TAK242和LY294002可以逆转上述效果。
结论:我们的研究结果表明,KJG可以通过抑制TLR4激活,通过PI3K/AKT/FOXO1通路调节神经炎症,发挥抗抑郁作用。这项研究的发现揭示了KJG抗抑郁作用的新机制,为抑郁症的靶向治疗方法的发展提供了有希望的途径。
BACKGROUND: Kaixin Jieyu Granule (KJG), an improved formula of Kai-xin-san and Si-ni-san, is a highly effective formula with demonstrated efficacy in preventing depression in previous studies. However, the underlying molecular mechanisms of KJG\'s antidepressant effects on inflammatory molecules remain unclear. This
study aimed to explore the therapeutic effects of KJG on depression using network pharmacology and experimental validation.
METHODS: We employed a multi-faceted approach, combining high-performance liquid chromatography (HPLC), network pharmacology, and molecular docking, to unravel the underlying mechanisms of KJG\'s anti-depressant effects. To confirm our findings, we conducted at least two independent in vivo experiments on mice, utilizing both the chronic unpredictable mild stress (CUMS)-induced and lipopolysaccharide (LPS)-induced models. Furthermore, the results of in vivo experiments were verified by in vitro assays. Behavioral tests were utilized to evaluate depression-like behaviors, while Nissl staining was used to assess morphological changes in the hippocampus. Pro-inflammatory cytokines and pathway-related protein expressions were determined using a combination of immunofluorescence staining, enzyme-linked immunosorbent assay (ELISA), and Western Blotting (WB).
RESULTS: Our network-based approaches indicated that ginsenoside Rg1 (GRg1) and saikosaponin d (Ssd) are the major constituents of KJG that exert an anti-depressant effect by regulating TLR4, PI3K, AKT1, and FOXO1 targets through the toll-like receptor, PI3K/AKT, and FoxO pathways. In vivo, KJG can attenuate depression-like behaviors, protect hippocampal neuronal cells, and reduce the production of pro-inflammatory mediators (TNF-α, IL-6, and IL-1β) by repressing TLR4 expression, which was regulated by the inhibition of FOXO1 through nuclear exportation. Furthermore, KJG increases the expression levels of PI3K, AKT, p-PI3K, p-AKT, and p-PTEN. Our in vitro assays are consistent with our in vivo studies. On the other hand, the above effects can be reversed by applying TAK242 and LY294002.
CONCLUSIONS: Our findings suggest that KJG can exert anti-depressant effects by regulating neuroinflammation through the PI3K/AKT/FOXO1 pathway by suppressing TLR4 activation. The
study\'s findings reveal novel mechanisms underlying the anti-depressant effects of KJG, presenting promising avenues for the development of targeted therapeutic approaches for depression.