Objective To investigate the mechanism of Longdan Xiegan Tang (LXT) in the treatment of diabetic peripheral neuralgia (DPN).

Methods Network pharmacology was used to predict the pathway targets of LXT in the treatment of DPN, and molecular docking was performed between its major active components and the key target serine/threonine protein kinase 1 (AKT1). A DPN model was established in female C57BL mice using a high-fat diet combined with intraperitoneal injection of streptozotocin (STZ). The mice were divided into a blank group, a control group, a duloxetine group, and an LXT group. The therapeutic effects of LXT on DPN mice were evaluated by measuring body weight and blood glucose, together with behavioral tests, including thermal withdrawal latency (TWL) and mechanical withdrawal threshold (MWT). In vitro, under a high-glucose (HG) microenvironment, a CCK-8 cell viability assay was used to analyze proliferative activity following intervention with a PI3K agonist (740Y-P), a PI3K antagonist (LY294002), an AKT agonist (SC79), an AKT antagonist (MK-2206), combined intervention with a PI3K antagonist plus an AKT agonist (HG+LY+SC), and combined intervention with an AKT agonist plus an AKT antagonist (HG+SC+MK). Flow cytometry was used to analyze the early apoptosis rate and viable-cell survival rate under the above interventions.

Results Network pharmacology analysis identified 70 effective active components of LXT and 194 intersecting targets with DPN, and screened five core active components and five core targets, which mainly regulate signaling pathways such as PI3K/AKT and AGE-RAGE. Molecular docking showed that the binding energy between the core target AKT1 and 7-hydroxy-5-methoxyflavone reached −6.42 kcal/mol, and AKT1 formed characteristic hydrogen bonds with four core components. Animal experiments verified successful establishment of the DPN model. After 14 consecutive days of treatment, both the LXT group and the duloxetine group showed improvements in TWL and MWT (both P ＜ 0.001). Cellular experiments showed that the proliferative activity of Schwann cells in the HG group was (62.766±1.922)%, which was lower than that in the low-glucose group (94.771±1.376)% (P ＜ 0.05). After intervention with agonists 740Y-P and SC79 proliferative activity increased to (67.016±1.284)% and (77.267±0.661)%, respectively (both P ＜ 0.05), whereas antagonists LY294002 and MK-2206 further aggravated the inhibition. Factorial analysis revealed interaction effects between SC79 and LY294002 (F = 57.82, P ＜ 0.0001) and between SC79 and MK-2206 (F = 36.53, P = 0.0003).

Conclusions LXT exerts a therapeutic effect in the treatment of experimental DPN. The PI3K/AKT signaling pathway predicted by network pharmacology was validated by cellular experiments to exert regulatory effects on apoptosis in Schwann cells, and PI3K and AKT show a cascade-dependent relationship, which may represent the mechanism of LXT in the treatment of DPN.