Targeting phenylpyruvate restrains excessive NLRP3 inflammasome activation and pathological inflammation in diabetic wound healing.
Moderate inflammation is essential for standard wound healing. In pathological conditions, such as diabetes, protracted and refractory wounds are associated with excessive inflammation, manifested by persistent proinflammatory macrophage states. However, the mechanisms are still unclear. Herein, we perform a metabolomic profile and find a significant phenylpyruvate accumulation in diabetic foot ulcers. Increased phenylpyruvate impairs wound healing and augments inflammatory responses, whereas reducing phenylpyruvate via dietary phenylalanine restriction relieves uncontrolled inflammation and benefits diabetic wounds. Mechanistically, phenylpyruvate is ingested into macrophages in a scavenger receptor CD36-dependent manner, binds to PPT1, and inhibits depalmitoylase activity, thus increasing palmitoylation of the NLRP3 protein. Increased NLRP3 palmitoylation is found to enhance NLRP3 protein stability, decrease lysosome degradation, and promote NLRP3 inflammasome activation and the release of inflammatory factors, such as interleukin (IL)-1β, finally triggering the proinflammatory macrophage phenotype. Our study suggests a potential strategy of targeting phenylpyruvate to prevent excessive inflammation in diabetic wounds.
Pubmed ID: 37480849 RIS Download
Research resources used in this publication
None foundAdditional research tools detected in this publication
- UniProt (RRID:SCR_002380)
- SPSS (RRID:SCR_002865)
- PeproTech (RRID:SCR_006802)
- FlowJo (RRID:SCR_008520)
- Proteintech Group (RRID:SCR_008986)
- Adobe Illustrator (RRID:SCR_010279)
- AdipoGen (RRID:SCR_010507)
- ProteoWizard (RRID:SCR_012056)
- AutoDock (RRID:SCR_012746)
- Abcam (RRID:SCR_012931)
- BD Biosciences (RRID:SCR_013311)
- MetaboLights (RRID:SCR_014663)
- XCMS (RRID:SCR_015538)
- Autogrid (RRID:SCR_015982)
- test (RRID:SCR_018743)
- ThermoFisher Scientific EVOS M5000 Imaging System (RRID:SCR_023650)
Antibodies used in this publication
- Goat Anti-Rat IgG(H+L), FITC conjugate (RRID:AB_2890989)
- Alexa Fluor 647-conjugated Goat Anti-Rabbit IgG (H+L) (RRID:AB_2768330)
- Alexa Fluor 555-conjugated Goat Anti-Mouse IgG (H+L) (RRID:AB_2768321)
- TruStain FcX(TM) (anti-mouse CD16/32) (RRID:AB_1574973)
- APC anti-mouse CD36 (RRID:AB_571994)
- APC anti-mouse/human CD11b (RRID:AB_312795)
- Brilliant Violet 421(TM) anti-mouse F4/80 (RRID:AB_2563102)
- PE anti-mouse CD163 (RRID:AB_2860724)
- APC anti-mouse CD86 (RRID:AB_493342)
- CD107b antibody (RRID:AB_2881468)
- CD206/MRC1 (E6T5J) XP® Rabbit mAb (RRID:AB_2892682)
- iNOS Rabbit Antibody (RRID:AB_2687529)
- Anti -F4/80 (RRID:AB_2814687)
- CD31 antibody (RRID:AB_2299349)
- smooth muscle actin antibody (RRID:AB_2223009)
- Ki67 antibody - Proliferation Marker (RRID:AB_443209)
- Low Density LRP antibody [EPR3724] (RRID:AB_2234877)
- Hsc70 antibody (RRID:AB_2120153)
- Mannose 6 Phosphate Receptor (Cation independent) antibody [EPR6599] (RRID:AB_10974087)
- Recombinant Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [EPR20018-251] (RRID:AB_2916341)
- Anti-PPT1 Antibody (RRID:AB_2923599)
- Anti-p38 (phospho T180 + Y182) antibody [EPR18120] (RRID:AB_2576214)
- AKT antibody (RRID:AB_10912803)
- Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (RRID:AB_2315049)
- Recombinant Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (RRID:AB_2934088)
- Recombinant Anti-ERK1 + ERK2 antibody [EPR17526] (RRID:AB_2802136)
- NFkB p65 antibody [E379] (RRID:AB_776751)
- NFkB p65 (phospho S536) antibody [EP2294Y] (RRID:AB_1524028)
- Rabbit Anti-GAPDH Monoclonal Antibody, Unconjugated, Clone 14C10 (RRID:AB_561053)
- β-Actin (13E5) Rabbit mAb (RRID:AB_2223172)
- Caspase-1 (E2Z1C) Rabbit mAb (RRID:AB_2890194)
- Cleaved Caspase-1 (Asp296) (E2G2I) Rabbit mAb (RRID:AB_2923067)
- IL-1β (D6D6T) Rabbit mAb (Mouse Specific) (RRID:AB_2799001)
- Cleaved-IL-1β (Asp117) (E7V2A) Rabbit mAb (Mouse Specific) (RRID:AB_2799639)
- ASC/TMS1 (D2W8U) Rabbit mAb (Mouse Specific) (RRID:AB_2799736)
- anti-NLRP3/NALP3, mAb (Cryo-2) (RRID:AB_2885199)
- NLRP3 (D4D8T) (RRID:AB_2722591)
- NLRP3 Antibody (RRID:AB_2839376)
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This is a list of tools and resources that we have found mentioned in this publication.
UniProt (tool)
RRID:SCR_002380
Collection of data of protein sequence and functional information. Resource for protein sequence and annotation data. Consortium for preservation of the UniProt databases: UniProt Knowledgebase (UniProtKB), UniProt Reference Clusters (UniRef), and UniProt Archive (UniParc), UniProt Proteomes. Collaboration between European Bioinformatics Institute (EMBL-EBI), SIB Swiss Institute of Bioinformatics and Protein Information Resource. Swiss-Prot is a curated subset of UniProtKB.
View all literature mentionsSPSS (tool)
RRID:SCR_002865
Software package used for interactive, or batched, statistical analysis in social science, health sciences and marketing. Software platform offers advanced statistical analysis, a library of machine-learning algorithms, text analysis, open-source extensibility, integration with big data and deployment into applications.Versions that were produced by SPSS Inc. before the IBM acquisition (Versions 18 and earlier) would be given origin or publisher of SPSS Inc. in Chicago.
View all literature mentionsFlowJo (tool)
RRID:SCR_008520
Software for single-cell flow cytometry analysis. Its functions include management, display, manipulation, analysis and publication of the data stream produced by flow and mass cytometers.
View all literature mentionsProteintech Group (tool)
RRID:SCR_008986
Proteintech Europe Ltd is an ISO 9001:2008 certified company
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RRID:SCR_010279
Vector graphics software to create digital graphics, illustrations, and typography for several types of media: print, web, interactive, video, and mobile.
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RRID:SCR_012056
Software that enables rapid tool creation by providing a robust, pluggable development framework that simplifies and unifies data file access, and performs standard proteomics and LCMS dataset computations.
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RRID:SCR_012746
Software suite of automated docking tools. Designed to predict how small molecules, such as substrates or drug candidates, bind to receptor of known 3D structure. AutoDock consist of AutoDock 4 and AutoDock Vina. AutoDock 4 consists of autodock to perform docking of ligand to set of grids describing target protein, and autogrid to pre calculate these grids.
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RRID:SCR_012931
A commercial antibody supplier which supplies primary and secondary antibodies, biochemicals, proteins, peptides, lysates, immunoassays and other kits.
View all literature mentionsMetaboLights (tool)
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A cross-species, cross-technique database for metabolomics experiments, data, and derived information. It includes metabolite structures and their reference spectra, their biological roles, locations and concentrations, and experimental data from metabolic experiments.
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RRID:SCR_015538
Cloud-based mass spectrometry data processing platform for metabolomics and lipidomics.
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RRID:SCR_015982
THIS RESOURCE IS NO LONGER IN SERVICE. Documented on February 23,2023.Software for automated docking analysis to precalculate the set of grids describing the target protein. It is a part of automated molecular modeling simulation software AutoDock.
View all literature mentionsThermoFisher Scientific EVOS M5000 Imaging System (tool)
RRID:SCR_023650
Microsoft system includes integrated digital inverted benchtop microscope for four-color fluorescence, transmitted-light, and color imaging. Provides interchangeable optics with autofocus, single-click multi-channel image acquisition.3.2 Megapixels, monochrome, CMOS camera. Offers software for acquisition, analysis, and automated cell counting.
View all literature mentionsGoat Anti-Rat IgG(H+L), FITC conjugate (antibody)
RRID:AB_2890989
This polyclonal secondary targets IgG
View all literature mentionsAlexa Fluor 647-conjugated Goat Anti-Rabbit IgG (H+L) (antibody)
RRID:AB_2768330
This unknown targets IgG (H+L)
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RRID:AB_2768321
This unknown targets IgG (H+L)
View all literature mentionsTruStain FcX(TM) (anti-mouse CD16/32) (antibody)
RRID:AB_1574973
This monoclonal targets CD16/32
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RRID:AB_571994
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RRID:AB_312795
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RRID:AB_2860724
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View all literature mentionsAPC anti-mouse CD86 (antibody)
RRID:AB_493342
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View all literature mentionsCD107b antibody (antibody)
RRID:AB_2881468
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View all literature mentionsCD206/MRC1 (E6T5J) XP® Rabbit mAb (antibody)
RRID:AB_2892682
This monoclonal targets CD206/MRC1
View all literature mentionsiNOS Rabbit Antibody (antibody)
RRID:AB_2687529
This monoclonal targets iNOS
View all literature mentionssmooth muscle actin antibody (antibody)
RRID:AB_2223009
This polyclonal targets smooth muscle actin
View all literature mentionsKi67 antibody - Proliferation Marker (antibody)
RRID:AB_443209
This polyclonal targets Ki67 antibody - Proliferation Marker
View all literature mentionsLow Density LRP antibody [EPR3724] (antibody)
RRID:AB_2234877
This monoclonal targets LRP1
View all literature mentionsHsc70 antibody (antibody)
RRID:AB_2120153
This polyclonal targets Hsc70
View all literature mentionsMannose 6 Phosphate Receptor (Cation independent) antibody [EPR6599] (antibody)
RRID:AB_10974087
This monoclonal targets Mannose 6 Phosphate Receptor (Cation independent) antibody [EPR6599]
View all literature mentionsRecombinant Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [EPR20018-251] (antibody)
RRID:AB_2916341
This recombinant monoclonal targets DDDDK epitope tag
View all literature mentionsAnti-PPT1 Antibody (antibody)
RRID:AB_2923599
This polyclonal targets PPT1
View all literature mentionsAnti-p38 (phospho T180 + Y182) antibody [EPR18120] (antibody)
RRID:AB_2576214
This monoclonal targets phospho-p38 (phospho T180 + Y182)
View all literature mentionsPhospho-Akt (Ser473) (D9E) XP® Rabbit mAb (antibody)
RRID:AB_2315049
This monoclonal targets Phospho-Akt (Ser473)
View all literature mentionsRecombinant Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (antibody)
RRID:AB_2934088
This recombinant monoclonal targets ERK1 (phospho T202) + ERK2 (phospho T185)
View all literature mentionsRecombinant Anti-ERK1 + ERK2 antibody [EPR17526] (antibody)
RRID:AB_2802136
This recombinant targets ERK1 + ERK2
View all literature mentionsNFkB p65 antibody [E379] (antibody)
RRID:AB_776751
This monoclonal targets Human NFkB p65
View all literature mentionsNFkB p65 (phospho S536) antibody [EP2294Y] (antibody)
RRID:AB_1524028
This monoclonal targets Human NFkB p65 (phospho S536)
View all literature mentionsRabbit Anti-GAPDH Monoclonal Antibody, Unconjugated, Clone 14C10 (antibody)
RRID:AB_561053
This recombinant monoclonal targets GAPDH
View all literature mentionsβ-Actin (13E5) Rabbit mAb (antibody)
RRID:AB_2223172
This monoclonal targets beta-Actin
View all literature mentionsCaspase-1 (E2Z1C) Rabbit mAb (antibody)
RRID:AB_2890194
This monoclonal targets Caspase-1
View all literature mentionsCleaved Caspase-1 (Asp296) (E2G2I) Rabbit mAb (antibody)
RRID:AB_2923067
This monoclonal targets Cleaved Caspase-1
View all literature mentionsIL-1β (D6D6T) Rabbit mAb (Mouse Specific) (antibody)
RRID:AB_2799001
This monoclonal targets IL1B
View all literature mentionsCleaved-IL-1β (Asp117) (E7V2A) Rabbit mAb (Mouse Specific) (antibody)
RRID:AB_2799639
This monoclonal targets IL1B
View all literature mentionsASC/TMS1 (D2W8U) Rabbit mAb (Mouse Specific) (antibody)
RRID:AB_2799736
This monoclonal targets PYCARD
View all literature mentionsanti-NLRP3/NALP3, mAb (Cryo-2) (antibody)
RRID:AB_2885199
This monoclonal targets NLRP3/NALP3
View all literature mentions
