MetaboLights

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identification of metabolites was performed by comparing of accuracy m/z value (<10 ppm), and MS/MS spectra with an in-house database established with available authentic standards.All annotations were subjected to strict manual secondary verification and confirmation, with identification confidence level at Level 2 or above according to MSI criteria.MetaboLightsPublicLiquid Chromatography MS - negative - reverse-phaseLiquid Chromatography MS - positive - reverse-phaseAnalysis was performed using an UHPLC (1290 Infinity LC, Agilent Technologies) coupled to a quadrupole time-of-flight (AB Sciex TripleTOF 6600). For HILIC separation, samples were analyzed using a 2.1 mm ¡Á 100 mm ACQUITY UPLC BEH Amide 1.7 ¦Ìm column (waters, Ireland). In both ESI positive and negative modes, the mobile phase contained A = 25 mM ammonium acetate and 25 mM ammonium hydroxide in water and B = acetonitrile. The gradient was 95% B for 0.5 min and was linearly reduced to 65% in 6.5 min, and then was reduced to 40% in 1 min and kept for 1 min, and then increased to 95% in 0.1 min, with a 3 min re-equilibration period.Frontiers in Immunology.Liaoning University Of Traditional Chinese MedicineYOU LINXINblood serummass spectrometry assayRat serum samples were thawed at 4¡ãC. An aliquot of each sample was mixed with pre-cooled methanol/acetonitrile/water (2:2:1, v/v/v), vortexed, and sonicated at low temperature for 30 min. After incubation at ?20¡ãC for 10 min, the mixture was centrifuged at 14,000 g for 20 min at 4¡ãC. The supernatant was dried in a vacuum centrifuge. For LC-MS analysis, the samples were re-dissolved in 100 ¦ÌL acetonitrile/water (1:1, v/v) solvent and centrifuged at 14,000 g at 4¡ãC for 15 min, then the supernatant was injected.Rattus norvegicushttps://www.ebi.ac.uk/metabolights/MTBLS14327JingYu Zhang. 1711270021@qq.com.SiJia Li. lnzyydxlsj@163.com.JiQuan Li. Liaoning University Of Traditional Chinese Medicine. lnzy_zjtn_ljq@126.com.Yan Zhao. 723891308@qq.com.LinXin You. youlinxin282520@163.com.The raw MS data were converted to MzXML files using ProteoWizard MSConvert before importing into freely available XCMS software. For peak picking, the following parameters were used: centWave m/z = 10 ppm, peakwidth = c (10, 60), prefilter = c (10, 100). For peak grouping, bw = 5, mzwid = 0.025, minfrac = 0.5 were used. CAMERA (Collection of Algorithms of MEtabolite pRofile Annotation) was used for annotation of isotopes and adducts. In the extracted ion features, only the variables having more than 50% of the nonzero measurement values in at least one group were kept. After sum-normalization, the processed data were analyzed by R package (ropls), where it was subjected to multivariate data analysis, including Pareto-scaled principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA). The 7-fold cross-validation and response permutation testing were used to evaluate the robustness of the model. The variable importance in the projection (VIP) value of each variable in the OPLS-DA model was calculated to indicate its contribution to the classification. Student¡¯s t test was applied to determine the significance of differences between two groups of independent samples. VIP > 1 and p value < 0.05 were used to screen significant changed metabolites. Pearson¡¯s correlation analysis was performed to determine the correlation between two variables.GroupTreatmentyoulinxin282520@163.comBlood was collected without anticoagulant and allowed to clot at room temperature for 30 min, then the samples were centrifuged for 15 min (1,500 g, 4¡ãC). Serum was obtained and stored at ?80¡ãC until LC-MS analysis.MetabolomicsMetabolomicsElectroacupunctureuntargeted analysisRattus norvegicusblood serumurticariaLipid Metabolismexperimental blankComplement systemAgilent 1290 Infinity HPLCAmino acid metabolismAB SCIEX TripleTOF 6600MetabolomicsElectroacupunctureuntargeted analysisurticariaRattus norvegicusblood serumLipid Metabolismexperimental blankComplement systemAgilent 1290 Infinity HPLCAmino acid metabolismAB SCIEX TripleTOF 6600The ESI source conditions were set as follows: Ion Source Gas1 (Gas1) as 60, Ion Source Gas2 (Gas2) as 60, curtain gas (CUR) as 30, source temperature: 600¡ãC, IonSpray Voltage Floating (ISVF) ¡À 5500 V. In MS only acquisition, the instrument was set to acquire over the m/z range 60-1000 Da, and the accumulation time for TOF MS scan was set at 0.20 s/spectra. In auto MS/MS acquisition, the instrument was set to acquire over the m/z range 25-1000 Da, and the accumulation time for product ion scan was set at 0.05 s/spectra. The product ion scan is acquired using information dependent acquisition (IDA) with high sensitivity mode selected. The parameters were set as follows: the collision energy (CE) was fixed at 35 V with ¡À 15 eV; declustering potential (DP), 60 V (+) and ?60 V (?); exclude isotopes within 4 Da, candidate ions to monitor per cycle: 10.falseElectroacupuncture at Ll11 and SP10 is associated with alleviation of acute urticaria-like reactions in passive cutaneous anaphylaxis: an exploratory analysis of complement-related proteins and multiscale omicsBackground: 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.2026-06-032026-04-21MTBLS14327