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TMT-based proteomic analysis of electroacupuncture and drug intervention in OVA-induced urticaria-like passive cutaneous anaphylaxis: rat model

ABSTRACT: This project investigates protein expression changes in rat skin tissue following electroacupuncture (EA) treatment and positive drug intervention in an OVA-induced passive cutaneous anaphylaxis (PCA) urticaria-like model. A total of 12 skin samples were divided into 4 groups: Control (K), Model (M), EA treatment (X), and Positive drug control (Z), with 3 biological replicates per group. Proteins were labeled using TMT reagents and analyzed by LC-MS/MS (Q-Exactive). Differential expression analysis (FC>1.2, p<0.05) was performed for comparisons including M_vs_K, Z_vs_M, X_vs_M, and X_vs_Z.

ORGANISM(S): Rattus Norvegicus

SUBMITTER: Jiquan Li

PROVIDER: PXD075803 | iProX | Wed Mar 18 00:00:00 GMT 2026

REPOSITORIES: iProX

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Project description:Background: The lifetime prevalence of acute urticaria is approximately 20%. Although electroacupuncture (EA) has been investigated as an adjunctive treatment for urticaria, the molecular mechanisms underlying these effects remain elusive. Passive cutaneous anaphylaxis (PCA) serves as an experimental model of localized acute IgE-mediated hypersensitivity in the skin, and it recapitulates the key phenotypic characteristics of acute urticaria. In this study, we leverage an ovalbumin (OVA)-induced PCA model to characterize molecular alterations in the skin and serum following EA intervention at LI11 and SP10 using integrative proteomic and metabolomic analyses.Methods: An acute urticaria-like model was established via OVA-induced PCA. Rats were treated at the relevant acupoints for 5 consecutive days, during which the blue wheal areas and scratching frequencies were recorded. Immunoassays (ELISA and immunohistochemistry) and histological staining (H&E and toluidine blue) were used to assess cutaneous pathological alterations, vascular permeability, and inflammatory status. Proteomic and metabolomic analyses were conducted on skin and serum samples from three experimental groups to identify differentially expressed proteins and metabolites. An integrative analysis of the omics data was then conducted to explore potential molecular indicators associated with this intervention at both the protein and metabolite levels.Results: In the acute PCA model, the treatment was concurrent with reduced wheal areas, decreased scratching frequencies, and diminished cutaneous vascular permeabilities. These results suggested attenuated acute inflammatory responses. Multi-omics profiling revealed alterations in the lipid and amino acid metabolic pathways linked to this intervention. The western blot and ELISA results indicated that this intervention coincided with the decreased expression of C3a/C5a and their receptors (C3aR/C5aR) in skin tissues, as well as reduced serum levels of inflammatory mediators (IL-6, TNF-a, and 5-HT).Conclusions: The results of our study indicated that in the OVA-induced PCA model, this intervention was associated with decreased expression of components of the C3a/C5a-C3aR/C5aR axis and alterations in metabolic profiles. These exploratory findings may inform future mechanistic studies of complement and metabolism associations in this acute model.

Project description:The electroacupuncture-induced analgesic effect has been used widely to alleviate diverse pains. However, significant individual variations in analgesic effect of EA for both experiments and clinics were reported. According to the sensitivity of the analgesic response to EA stimulation, the subjects could be categorized into high responders (HR) and low responders (LR). However, the molecular mechanism of individual variability in the analgesic response to acupuncture stimulation is still uncertain. This study aims to investigate the potential gene expression in spinal dorsal horn induced by 2Hz/100Hz electroacupuncture in HR and LR rats. Rats were given 2Hz or 100Hz electroacupuncture for 30 min and using cDNA microarrays to compare different gene expression in dorsal horn. Transcriptome profiling analysis found that some co-regulated genes related with electroacupuncture or 2Hz or 100Hz freqencies. These co-regulated genes were plasticity-related by GO analysis. We also found some special regulated genes in HR vs. LR in 2Hz/100Hz electroacupuncture stimulation. These results suggested that neurotransmitter system and cytokine different between HR with LR in 2Hz electroacupuncture. But in 100Hz electroacupunture, there were many different regulated genes related with ribosome between HR with LR, which needs more studies to research the function and may play an important role in HR vs. LR by 100Hz electroacupuncture. Keywords: Transcriptome analysis Rats were exposed to different frequencies (2Hz or 100Hz) electroacupuncture stimlation for 30 min and nociceptive testing and returned to home cages for 1 hours before sacrificed. According to the sensitivity of the analgesic response to 2Hz or 100Hz EA stimulation, the rats divided into four groups: 2Hz-HR group, 2Hz-LR group, 100Hz-HR group and 100Hz-LR group. Subsequently analyzed their dorsal horn transcript profile using cDNA microarrays, the rats were without receiving electroacupuncture as control. The tissue of dorsal horn (DH) of the fifth and sixth lumbar (L5 and L6) of four or five rats were selected for transcript analysis in each group. The five control rats were mixed and labeled with cy5, each rat of experiment groups was labeled with cy3.

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TMT-based proteomic analysis of electroacupuncture and drug intervention in OVA-induced urticaria-like passive cutaneous anaphylaxis: rat model

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