背景:金葵肾气丸(JSP)是一种用于治疗“肾阳虚”疾病的经典中药。先前的研究表明JSP对小鼠神经元凋亡的保护作用。
目的:本研究,将网络药理学与体内实验相结合,探讨JSP预防小鼠神经管缺陷(NTDs)的作用机制。
方法:网络药理学分析了JSP成分和靶标,鉴定具有NTDs的常见基因并探索潜在途径。分子对接评估关键JSP组分和途径蛋白之间的相互作用。在全反式维甲酸(atRA)诱导的NTDs小鼠模型中,HE染色观察组织病理学改变,使用TUNEL检测神经元凋亡,和WesternBlot评估PI3K/AKT信号通路和凋亡相关蛋白的变化。
结果:不同浓度的JSP导致小鼠胚胎中神经管缺陷发生的不同程度的减少,最高剂量显示出最显著的下降。此外,与叶酸(FA)相比,它显示出更好的NTDs降低率。网络药理学构建了药物-活性成分-基因靶网络,建议使用关键活性成分,例如槲皮素,Wogonin,β-谷甾醇,山奈酚,和豆甾醇,可能作用于PI3K/Akt信号通路。分子对接证实了稳定的结合结构。Western印迹分析显示p-PI3K的表达增加,p-Akt,p-Akt1,p-Akt2,p-Akt3,裂解的半胱天冬酶-3和Bax的下调,和上调Bcl-2,表明通过抗凋亡作用预防NTDs。
结论:我们已经确定了JSP预防NTDs的有效剂量,在atRA诱导的小鼠胚胎NTDs中,通过激活PI3K/Akt信号通路和抑制细胞凋亡揭示其潜力。
BACKGROUND: jinkui Shenqi Pill (JSP) is a classic traditional Chinese medicine used to treat \"Kidney Yang Deficiency\" disease. Previous studies indicate a protective effect of JSP on apoptosis in mouse neurons.
OBJECTIVE: This research, combining network pharmacology with in vivo experiments, explores the mechanism of JSP in preventing neural tube defects (NTDs) in mice.
METHODS: Network pharmacology analyzed JSP components and targets, identifying common genes with NTDs and exploring potential pathways. Molecular docking assessed interactions between key JSP components and pathway proteins. In an all-trans retinoic acid (atRA)-induced NTDs mouse model, histopathological changes were observed using HE staining, neuronal apoptosis was detected using TUNEL, and Western Blot assessed changes in the PI3K/AKT signaling pathway and apoptosis-related proteins.
RESULTS: Different concentrations of JSP led to varying degrees of reduction in the occurrence of neural tube defects in mouse embryos, with the highest dose showing the most significant decrease. Furthermore, it showed a better reduction in NTDs rates compared to folic acid (FA). Network pharmacology constructed a Drug-Active Ingredient-Gene Target network, suggesting key active ingredients such as Quercetin, Wogonin, Beta-Sitosterol, Kaempferol, and Stigmasterol, possibly acting on the PI3K/Akt signaling pathway. Molecular docking confirmed stable binding structures. Western Blot analysis demonstrated increased expression of p-PI3K, p-Akt, p-Akt1, p-Akt2, p-Akt3, downregulation of cleaved caspase-3 and Bax, and upregulation of Bcl-2, indicating prevention of NTDs through anti-apoptotic effects.
CONCLUSIONS: We have identified an effective dosage of JSP for preventing NTDs, revealing its potential by activating the PI3K/Akt signaling pathway and inhibiting cell apoptosis in atRA-induced mouse embryonic NTDs.