Neuropathic pain (NP) is a type of chronic pain resulting from injury or dysfunction of the nerves or spinal cord. Previous studies have shown that the combination of ligustrazine (LGZ) and sinomenine (SIN) exerts a synergistic antinociceptive effect in peripheral and central NP models. On this basis, a comprehensive analgesic evaluation was performed in a chronic constriction injury (CCI)-induced NP model in rats. Sciatic nerve histopathological changes were observed, and 22 cytokines and chemokines levels were analyzed. We also combined network pharmacology and metabolomics to explore their molecular mechanisms. Results showed that the combination of LGZ and SIN significantly alleviated the pain-like behaviors in CCI rats in a time- and dose-dependent manner, demonstrating superior therapeutic effects compared to LGZ or SIN alone. It also improved pathological damage to sciatic nerves and regulated inflammatory cytokine levels. Network pharmacology identified shared and distinct pain-related targets for LGZ and SIN, while metabolomics revealed 54 differential metabolites in plasma, and 17 differential metabolites in CSF were associated with the combined intervention of LGZ and SIN. Finally, through an integrated analysis of the core targets and differential metabolites, tyrosine metabolism, phenylalanine metabolism, and arginine and proline metabolism were identified as potential key metabolic pathways underlying the therapeutic effects of LGZ and SIN in CCI treatment. In conclusion, our study provides evidence to support the clinical application of LGZ and SIN in the treatment of NP.

Background: Neuropathic pain (NP) is a chronic pain condition caused by a lesion or disease of the somatosensory nervous system. Its development and persistence are closely associated with peripheral nociceptive input, central sensitization, neuroimmune dysregulation, and remodeling of the local metabolic microenvironment. Our previous studies demonstrated that combined treatment with ligustrazine and sinomenine ameliorates early sciatic nerve pathological damage and systemic metabolic disturbances in rats with chronic constriction injury (CCI). However, whether this combination modulates central sensitization in NP remains unclear. Objective: This study aimed to evaluate the effects of combined ligustrazine and sinomenine treatment on pain-related behaviors and motor function during the maintenance phase of CCI-induced neuropathic pain and to investigate the potential mechanisms underlying its attenuation of central sensitization, with particular emphasis on metabolic remodeling, neurotransmitter homeostasis, and associated regulatory molecules in the spinal dorsal horn. Methods: A rat model of CCI was established, and animals were treated with ligustrazine combined with sinomenine (12.5 + 12.5, 25 + 25, or 50 + 50 mg/kg) for 7 consecutive days. Mechanical withdrawal threshold, cold hypersensitivity, and gait behavior were assessed. A multiplex assay was used to profile 12 plasma inflammatory cytokines and pain-related mediators. Integrated transcriptomic and metabolomic analyses of the spinal dorsal horn were subsequently performed to identify key altered pathways. Oxylipin profiling, targeted neurotransmitter analysis, and quantification of metabolites associated with the arginine–polyamine pathway were further conducted to characterize alterations in the local metabolic microenvironment associated with central sensitization. Finally, quantitative real-time PCR (qRT-PCR) and Western blotting were used to evaluate molecules related to polyamine metabolism, redox homeostasis, and central sensitization. Results: Combined ligustrazine and sinomenine treatment improved pain-related behaviors and gait dysfunction in CCI rats and partially normalized systemic inflammatory dysregulation. The ipsilateral-to-contralateral maximum contact-area ratio decreased from 102.8 ± 9.0% in the Sham group to 53.1 ± 9.7% in the Model group and recovered to 85.5 ± 4.4% in the high-dose group. Metabolomic and transcriptomic analyses identified 54 differential metabolites and 130 differentially expressed genes, respectively, following treatment. Integrated multi-omics analysis highlighted arginine-related metabolic pathways, together with alterations in neurotransmitter, polyamine, oxylipin, and redox homeostasis.

Polygonum capitatum (PC) is an ethnomedicine with reported antibacterial and anti-inflammatory activities and has been clinically used in urinary tract infection (UTI)-related disorders. However, its in vivo exposure characteristics and metabolically associated therapeutic mechanisms in acute pyelonephritis (AP) remain insufficiently understood. To address this issue, this study aimed to evaluate the therapeutic effects of PC in an Escherichia coli (E. coli)-induced rat model of AP and to explore constituents and metabolic pathways associated with its activity. Different PC extracts were screened for antibacterial and anti-inflammatory activities, and the 70% ethanol extract was selected for further study. Seven major compounds were quantified by HPLC. In AP rats, the pharmacokinetic profiles of these compounds in plasma and the renal cortex were analyzed by microdialysis-coupled HPLC-MS/MS. Pharmacodynamic evaluation included urinary bacterial load, urinalysis, renal function, inflammatory cytokines, and renal histopathology. Exploratory PK–PD analysis, untargeted renal metabolomics, and targeted metabolomics of the tryptophan–kynurenine (Trp–Kyn) pathway were also performed. The 70% ethanol extract of PC exhibited the strongest antibacterial and anti-inflammatory activities. The total content of seven active compounds was 3.85%, with gallic acid being the predominant compound (3.42%). Pharmacokinetic analysis revealed that gallic acid, protocatechuic acid, methyl gallate, and quercitrin achieved relatively high systemic exposure and renal distribution. In AP rats, the pharmacokinetic profiles of several compounds were altered, with increased plasma exposure of protocatechuic acid, vanillic acid, ethyl gallate, and syringic acid, while protocatechuic acid also showed higher exposure in the renal cortex. PC treatment reduced urinary bacterial load, improved renal function and urinalysis parameters, alleviated histopathological injury, and decreased inflammatory mediator levels, particularly in renal tissue. Exploratory PK–PD correlations were observed between several compounds and selected pharmacodynamic indicators. Metabolomic analysis suggested disturbances in glycerophospholipid metabolism and the Trp–Kyn pathway in AP rats, some of which were partially reversed after PC treatment. PC showed antibacterial and anti-inflammatory effects in AP rats. Gallic acid, protocatechuic acid, methyl gallate, and quercitrin may be candidate constituents associated with the therapeutic effects of PC, while modulation of glycerophospholipid metabolism and the Trp–Kyn pathway may be involved in its action against AP. These findings provide preclinical pharmacological evidence supporting the therapeutic potential of PC in AP.

Neuropathic pain (NP) is a chronic pain resulting from injury or dysfunction of the nerves or spinal cord. Previous studies have shown that the combination of ligustrazine (LGZ) and sinomenine (SIN) exerts a synergistic antinociceptive effect in peripheral and central NP models. This study aims to evaluate the pharmacodynamic characteristics of LGZ and SIN in a chronic constriction injury (CCI)-induced NP models in rats, and to analyze its molecular regulatory mechanisms from the perspectives of network pharmacology and metabolomics. Firstly, a comprehensive analgesic evaluation including the mechanical withdrawal threshold test, cold allodynia test, and the incapacitance test was performed after 1, 2 and 3 days of intervention. The sciatic nerve histopathological changes were observed, and inflammatory factor analysis were conducted. Secondly, the pain-related targets of LGZ and SIN were systematically analyzed based on network pharmacology. The differential metabolites in the plasma and cerebrospinal fluid (CSF) were screened, and their content were quantified using LC-MS metabolomics technology. Finally, the key metabolic pathways with crucial role of LGZ and SIN in treating NP was identified from a joint analysis of the potential targets and the differential metabolites. The combination of LGZ and SIN significantly alleviated pain-like behaviors of CCI rats time- and dose-dependently, and the therapeutic effect was notably superior to that of LGZ or SIN alone. The combination of LGZ and SIN could improve pathological damage to the sciatic nerve and regulate the levels of inflammatory cytokine. Network pharmacology analysis revealed that LGZ and SIN have 6 shared pain-related targets, while LGZ have 16 distinct pain-related targets and SIN have 52 distinct pain-related targets. The metabolomics analyses revealed that 54 differential metabolites in plasma and 17 differential metabolites in CSF were associated with the combined intervention of LGZ and SIN. Finally, through integrated analysis of the core targets and differential metabolites, tyrosine metabolism, phenylalanine metabolism, and arginine and proline metabolism were identified as potential key metabolic pathways underlying the therapeutic effects of LGZ and SIN in CCI treatment. This study provides an integrated approach to elucidate the molecular mechanisms underlying the combined use of LGZ and SIN in the treatment of NP. Furthermore, the findings offer new experimental evidence to support the clinical application of LGZ and SIN in the treatment of NP.

Background: Chronic insomnia is an important global health concern, but its molecular heterogeneity remains incompletely understood. Objective biomarkers may help characterize chronic insomnia and its traditional Chinese medicine (TCM) syndrome patterns. Methods: We applied a Public Evidence-informed Prioritization–Multi-omics Discovery–Targeted Validation (PMT) framework integrating public transcriptomic datasets with serum profiles from healthy controls and patients with chronic insomnia classified as excess (Shi) or deficiency (Xu) syndrome. Analyses included transcriptomic data processing and machine learning-based gene prioritization, serum metabolomics and lipidomics, targeted neurotransmitter quantification, bile acid profiling, and pathway-level comparison across omics datasets. Results: Transcriptomic analysis and weighted gene co-expression network analysis identified a phenotype-associated module containing 178 differentially expressed genes (DEGs), and three machine learning methods prioritized 11 candidate genes. Compared with controls, the Shi group had 53 differential metabolites and 87 differential lipids, whereas the Xu group had 49 differential metabolites and 80 differential lipids. Serum analyses revealed syndrome-associated differences in N-acetylated amino acids, lipid species, neurotransmitter-related metabolites, and bile acids. Kyoto Encyclopedia of Genes and Genomes (KEGG)-based integrative analysis identified shared pathways linking transcriptomic alterations with serum metabolomic and lipidomic changes, highlighting pathway-level convergence across the three omics layers. Cross-validated receiver operating characteristic analyses showed variable discriminatory performance, generally stronger for insomnia identification than for differentiation between Shi and Xu syndromes. Conclusions: Using the PMT framework, this study delineated distinct gene–metabolite–lipid signatures of chronic insomnia and its major TCM syndromes, providing a molecular basis for objective diagnosis, syndrome differentiation, and precision intervention.