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Pre-operative exercise therapy triggers anti-inflammatory trained immunity of Kupffer cells through metabolic reprogramming

Nature Metabolism volume 3, pages 843–858 (2021)Cite this article

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Abstract

Pre-operative exercise therapy improves outcomes for many patients who undergo surgery. Despite the well-known effects on tolerance to systemic perturbation, the mechanisms by which pre-operative exercise protects the organ that is operated on from inflammatory injury are unclear. Here, we show that four-week aerobic pre-operative exercise significantly attenuates liver injury and inflammation from ischaemia and reperfusion in mice. Remarkably, these beneficial effects last for seven more days after completing pre-operative exercising. We find that exercise specifically drives Kupffer cells toward an anti-inflammatory phenotype with trained immunity via metabolic reprogramming. Mechanistically, exercise-induced HMGB1 release enhances itaconate metabolism in the tricarboxylic acid cycle that impacts Kupffer cells in an NRF2-dependent manner. Therefore, these metabolites and cellular/molecular targets can be investigated as potential exercise-mimicking pharmaceutical candidates to protect against liver injury during surgery.

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Fig. 1: Exercise attenuates hepatic I/R injury by altering the immune microenvironment.
Fig. 2: Exercise shifts KCs towards an anti-inflammatory transcriptomic profile.
Fig. 3: Exercise drives KCs to be an anti-inflammatory phenotype.
Fig. 4: Exercise promotes metabolic reprogramming in KCs.
Fig. 5: Exercise-induced HMGB1 activates itaconate/IRG1/NRF2 signalling in KCs.
Fig. 6: Exercise promotes an anti-inflammatory trained immunity in KCs.

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Data availability

We have deposited the scRNA-seq data associated with this study with the Gene Expression Omnibus under accession no. GSE173429. Source data are provided with this paper. All other data that support the findings of this study are available from the corresponding author upon request.

Code availability

The code for the single-cell RNA sequencing data is provided with the paper in the supplementary dataset.

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Acknowledgements

We thank X. Liao and J. Imam for their assistance in preparing the manuscript. This work was supported by a National Institutes of Health grant no. 2T32AI106704-06 to A.O., grant nos. R01-CA214865-01 and R01-GM95566-06 to A.T., grant no. R01-AI152044 to M.D., grant nos. R01-GM120496 and R01-GM135234 to H. Wen and grant no. R01-GM137203 and Joseph A. Patrick Research Fellowship in Transplantation to H.H.

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Authors and Affiliations

  1. Department of Surgery, The Ohio State University Wexner Medical Center, Columbus, OH, USA

    Hongji Zhang, Yujia Xia, Amblessed Onuma, Yu Wang, Jiayi He, Han Wang, Ahmad Hamad, Chengli Shen, Meihong Deng, Allan Tsung & Hai Huang

  2. Department of Surgery, University of Pittsburgh, Pittsburgh, PA, USA

    Tianmeng Chen, Jinghua Ren, Junru Wu & Meihong Deng

  3. Cellular and Molecular Pathology Program, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

    Tianmeng Chen

  4. Department of Surgery, Union Hospital, Huazhong University of Science and Technology, Wuhan, China

    Jinxiang Zhang

  5. Mass Spectrometry Core, Beth Israel Deaconess Medical Center, Department of Medicine, Harvard Medical School, Boston, MA, USA

    John M. Asara

  6. Division of Hematology, Department of Internal Medicine, The Ohio State University Wexner Medical Center, Columbus, OH, USA

    Gregory K. Behbehani

  7. Department of Microbial Infection and Immunity, Infectious Disease Institute, The Ohio State University Comprehensive Cancer Center, Columbus, OH, USA

    Haitao Wen & Meihong Deng

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Contributions

H.Z., M.D., A.T. and H.H. conceived and designed the study. H.Z., M.D. and H.H. developed the methodology. J.R., Y.X. and H.H. acquired the data (provided animals, acquired and managed patients, provided the facilities). H.Z., A.O., C.S., J.H., H.H., T.C., J.W., Y.W., H. Wang, J.Z., G.K.B. and J.M.A. analysed and interpreted the data (statistical analysis, biostatistics, computational analysis). H.Z., H. Wen, M.D., A.T. and H.H. wrote and reviewed the manuscript. J.R., J.H. and A.H. provided administrative, technical, or material support (that is, reporting or organizing data, constructing databases). M.D., A.T. and H.H. supervised the study.

Corresponding authors

Correspondence to Meihong Deng, Allan Tsung or Hai Huang.

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Peer review information Nature Metabolism thanks the anonymous reviewers for their contribution to the peer review of this work. Primary Handling Editor: Isabella Samuelson.

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Extended data

Extended Data Fig. 1 Pre-operative exercise reduces hepatic I/R injury and inflammation.

a, Liver damage were measured by serum AST level (P = 0.7606, P = 0.3610, P = 0.0555, P = 0.0549, P < 0.0001, P < 0.0001, P < 0.0001, P < 0.0001 between exercise and sedentary group from 1day to 16 weeks) and liver histological evaluation (Suzuki scores)) (P = 0.7464, P = 0.1898, P = 0.0932, P = 0.0441, P < 0.0001, P = 0.0002, P < 0.0001, P < 0.0001 between exercise and sedentary group from 1day to 16 weeks) from mice with sedentary or pre-operative exercise treatment for 1 day, or 1 to 16 weeks which subjected to hepatic I/R (n = 6 mice per group). b, Liver damage were measured by plasma ALT (P = 0.7548 in sham group, P < 0.0001 in I/R group) and AST levels (P = 0.2743 in sham group, P < 0.0001 in I/R group) from mice with sedentary or pre-operative exercise treatment for 4 weeks which subjected to hepatic I/R (Each dot represents one mouse, n = 7 mice in Sham group, n = 11 mice in I/R group). c, Liver damage were measured by liver histological evaluation (necrotic areas and Suzuki scores) (for necrotic areas, (P = 0.1228 in sham group, P < 0.0001 in I/R group), for Suzuki scores, (P = 0.8095 in sham group, P = 0.0018 in I/R group) from mice with sedentary or pre-operative exercise treatment for 4 weeks which subjected to hepatic I/R (Each dot represents one mouse, n = 6 mice per group). d, Serum IL-1Ra (P = 0.0001 in 4 weeks group, P = 0.0008 in 8 weeks group, P = 0.0005 in 12 weeks group, P = 0.0006 in 16 weeks group), IL-10 (P = 0.0008 in 4 weeks group, P = 0.0002 in 8 weeks group P = 0.0005 in 12 weeks group P = 0.0002 in 16 weeks group), IL-1β (P = 0.0004 in 4 weeks group, P = 0.0003 in 8 weeks group, P = 0.0002 in 12 weeks group, P = 0.0005 in 16 weeks group), IL-6 (P = 0.0015 in 4 weeks group, P = 0.0011 in 8 weeks group, P = 0.0007 in 12 weeks group, P = 0.0012 in 16 weeks group) and TNF-α (P = 0.0006 in 4 weeks group, P = 0.0011 in 8 weeks group, P = 0.0005 in 12 weeks group, P = 0.0002 in 16 weeks group)concentrations were measured by ELISA from mice with sedentary or pre-operative exercise treatment for 4, 8, 12 or 16 weeks which subjected to hepatic I/R (n = 6 mice per group). e, Serum IFN-α (P = 0.0854 in sham group, P = 0.04854 in I/R group), IFN-β (P = 03053 in sham group, P = 0.1154 in I/R group)and IFN-γ (P = 0.1531 in sham group, P = 0.0015 in I/R group) concentrations were measured by ELISA from mice with sedentary or pre-operative exercise treatment for 4 weeks at baseline (SHAM group) and subjected to hepatic I/R (Each dot represents one mouse, n = 6 mice per group). f, Serum CXCL1 (P < 0.0001 in sham group, P < 0.0001 in I/R group), CXCL2 (P = 0.0040 in sham group, P = 0.0001 in I/R group) and CXCL3 (P = 0.0071 in sham group, P < 0.0001 in I/R group) concentrations were measured by ELISA from mice with sedentary or pre-operative exercise treatment for 4 weeks at baseline (SHAM group) and subjected to hepatic I/R (Each dot represents one mouse, n = 6 mice per group). g, Liver damage were measured by serum AST level (P = 0.0127 between sedentary and 0 day after exercise group, P = 0.0001 between sedentary and 3 days after exercise group, P = 0.0019 between sedentary and 5 days after exercise group, P = 0.0082 between sedentary and 7 days after exercise group, p = 0.4524 between 0 day and 3 days group, P = 0.6955 between 0 day and 5 days group, P = 0.9063 between 0 day and 7 days group, P = 0.6701 between 3 days and 5 days group, P = 0.5088 between 3 days and 7 days group, P = 0.7702 between 5 days and 7 days group) from mice with sedentary or pre-operative exercise treatment for 4 weeks and resting for 0, 3, 5, or 7 days before hepatic I/R (Each dot represents one mouse, n = 11 in sedentary group, n = 7 in 0D after Exercise group, n = 15 in 3D after Exercise group, n = 9 in 5D after Exercise group, n = 7 in 7D after Exercise group). Graphs show the mean ± SD (a to f). Circles, triangles and squares represent individual mice. ANOVA with adjustment for multiple comparisons for (a and d). Unpaired two-sample Student’s t test for (b, c, e and f). NS: not significant. All the statistical test used in this figure was two-sided and two-sided P < 0.05 was considered statistically significant.

Extended Data Fig. 2 Mass cytometry and flow cytometry characterize hepatic immune profile.

a, Mass cytometry (CyTOF) of hepatic immune microenvironment profile from exercise or sedentary mice (4 weeks) with or without hepatic I/R in t-distributed stochastic neighbor embedding (t-SNE) graph. b, Percentage of Kupffer cells (KCs), monocytes, and neutrophils out of hepatic CD45 + cells. (Each dot represents one mouse, n = 2 mice per group) c, Percentage of neutrophils. d, Percentage of natural killer (NK) cells, NKT cells, and CD3 + T cells out of hepatic CD45 + cells in each group. e, Numbers of B cells (P = 0.3397 in sham group, P = 0.8025 in I/R group), CD4 + T cells (P = 0.6370 in sham group, P = 0.3695 in I/R group), CD8 + T cells(P = 0.7879 in sham group, P = 0.6170 in I/R group), and NKT cells(P = 0.6738 in sham group, P = 0.3010 in I/R group), dendritic cells (DCs) (P = 0.3910 in sham group, P = 0.7954 in I/R group), out of hepatic CD45 + cells in each group (Each dot represents one mouse, n = 6 mice per group). Circles and triangles represent individual mice. Graphs show the mean ± SD (b and e). Unpaired two-sample Student’s t test (b and e). All the statistical test used in this figure was two-sided and two-sided P < 0.05 was considered statistically significant.

Extended Data Fig. 3 Single cell RNA sequencing reveals transcriptomic profile of hepatic immune cells.

a, Experimental workflow of single cell RNA-sequencing in hepatic CD45 + cells with sedentary or pre-operative exercise treatment for 4 weeks. b, Heat map of gene expression profile in 15,332 hepatic immune cells clustered into 18 discrete cell populations. c, Transcriptomic profile of feature genes expression.

Extended Data Fig. 4 Single cell RNA sequencing reveals transcriptional signature of Kupffer cells.

a, Based on the transcriptomic profile, 15,332 cells were clustered into 18 discrete cell populations in Uniform Manifold Approximation and Projection (UMAP). b, Cluster 0 was one unique subpopulation of KCs mainly derived from exercise mice and Cluster 2 were mainly derived from sedentary mice. c, Kupffer cells percentages in cluster 0, 2, 5 and 10. d, Preference of cluster from Kupffer cell estimated by RO/E. + + + , RO/E > 2; ++, 1.5<RO/E < = 2; +/-, 0.5 = <RO/E < = 1.5; - RO/E < 0.5. e, Volcano plot shows that the expression of 6205 genes was significantly changed with pre-operative exercise.

Extended Data Fig. 5 Kupffer cells are required for pre-operative exercise mediated hepatic protection from I/R injury.

a, Experiment outline. b, Experiment outline. c, Flow cytometry characterization of Kupffer cells percentage after mice injected with saline or different concentration of GdCl3. d, Liver damage was measured by serum AST level (P = 0.0130 in saline group, P = 0.9197 in GdCl3 group) and liver histological evaluation (Suzuki scores) (P = 0.0023 in saline group, P = 0.2513 in GdCl3 group) from mice with sedentary or pre-operative exercise treatment for 4 weeks injected with GdCl3 (25 mg/kg) or saline solution intraperitoneally 48 hours before hepatic I/R (Each dot represents one mouse, n = 6 mice per group). e, Experiment outline. f, Adoptive transfer of Kupffer cells isolated from either sedentary or pre-operative exercise treatment for 4 weeks into Kupffer cells KO mice with GdCl3. Liver damage was measured by serum AST level (P = 0.0083 in sedentary recipient group, P = 0.0092 in exercise recipient group) and liver histological evaluation (Suzuki scores) (P = 0.0154 in sedentary recipient group, P = 0.0026 in exercise recipient group) among mice from these four groups subjected to hepatic I/R (Each dot represents one mouse, n = 6 mice per group). Graphs show the mean ± SD (d and f). Circles and triangles represent individual mouse. Unpaired two-sample Student’s t test for (d and f). All the statistical test used in this figure was two-sided and two-sided P < 0.05 was considered statistically significant.

Extended Data Fig. 6 Pre-operative exercise induces metabolic reprogramming in Kupffer cells.

a, Metabolite sets enrichment between Kupffer cells with sedentary or pre-operative exercise treatment for 4 weeks. b, Relative expression between Kupffer cells with sedentary or pre-operative exercise treatment for 4 weeks.

Extended Data Fig. 7 Pre-operative exercise-induced protective effect on hepatic I/R injury is dependent on itaconate/IRG1.

a, Experiment outline. b, Liver damage for Irg1+/+ and Irg1−/− mice with sedentary or pre-operative exercise treatment for 4 weeks after hepatic I/R was measured by serum AST level (P = 0.0013 between Irg1+/+ sedentary and Irg1+/+ exercise group, P = 0.0425 between Irg1+/+ sedentary and Irg1−/− sedentary group, P < 0.0001 between Irg1+/+ exercise and Irg1−/− sedentary group, P = 0.0005 between Irg1+/+ exercise and Irg1−/− exercise group, P = 0.7057 between Irg1+/+ sedentary and Irg1−/− exercise group) (Each dot represents one mouse, n = 6 mice per group). c, Serum IL-1Ra (P = 0.3521), IL-10 (P = 0.5245), IL-1β (P = 0.1919), IL-6 (P = 0.4139) and TNF-α (P = 0.9512) concentrations from Irg1−/− exercise or sedentary mice (4 weeks) after hepatic I/R were measured by ELISA (Each dot represents one mouse, n = 6 mice per group). d, Liver damage of KCs-depleted Irg1+/+ mice were adoptively transferred with Irg1−/− or Irg1+/+ KCs from sedentary or exercise mice after hepatic I/R were measured by serum AST levels ((P = 0.0020 between KC-Irg1+/+ sedentary and KC-Irg1+/+ exercise group, P = 0.0413 between KC-Irg1+/+ sedentary and KC-Irg1−/− sedentary group, P = 0.0004 between KC-Irg1+/+ exercise and KC-Irg1−/− sedentary group, P = 0.0003 between KC-Irg1+/+ exercise and KC-Irg1−/− exercise group, P = 0.6203 between KC-Irg1+/+ sedentary and KC-Irg1−/− exercise group) (Each dot represents one mouse, n = 6 mice per group). e, Liver damage for Irg1+/+ or Irg1−/− mice injected itaconate (ITA, 100 mg/kg) for 2 hours before hepatic ischaemia liver I/R and at the time of reperfusion with sedentary or pre-operative exercise treatment for 4 weeks after hepatic I/R was measured by serum AST level (P = 0.0028 between Irg1+/+ sedentary and Irg1+/+ exercise group, P = 0.0446 between Irg1+/+ sedentary and Irg1−/− sedentary group, P < 0.0001 between Irg1+/+ exercise and Irg1−/− sedentary group, P = 0.0802 between Irg1+/+ exercise and Irg1−/− exercise group, P = 0.0013 between Irg1−/− sedentary and Irg1−/− exercise group) (Each dot represents one mouse, n = 6 mice per group). f, Liver damage for Irg1+/+ or Irg1−/− mice injected 4-OI (25 mg/kg) for 24 and 2 hours before hepatic ischeamia, and at the time of reperfusion with sedentary or pre-operative exercise treatment for 4 weeks after hepatic I/R was measured by serum AST level (P = 0.0012 between Irg1+/+ sedentary and Irg1+/+ exercise group, P = 0.0070 between Irg1+/+ sedentary and Irg1−/− sedentary group, P < 0.0001 between Irg1+/+ exercise and Irg1−/− sedentary group, P = 0.0012 between Irg1+/+ exercise and Irg1−/− exercise group, P = 0.0027 between Irg1−/− sedentary and Irg1−/− exercise group) (Each dot represents one mouse, n = 6 mice per group) g, Liver damage for Nrf2+/+ and Nrf2−/− mice with sedentary or pre-operative exercise treatment for 4 weeks after hepatic I/R was measured by serum AST level (P = 0.0131 between Nrf2+/+ sedentary and Nrf2+/+ exercise group, P = 0.0486 between Nrf2+/+ sedentary and Nrf2−/− sedentary group, P = 0.0004 between Nrf2+/+ exercise and Nrf2−/− exercise group, P = 0.9587 between Nrf2−/− sedentary and Nrf2−/− exercise group) (Each dot represents one mouse, n = 3-6 mice in both Nrf2+/+ Sedentary and Exercise group, n = 3 in Nrf2−/− Sedentary group, n = 5 in Nrf2−/− Exercise group). Graphs show the mean ± SD (b to g). Circles, triangles and squares represent individual mice. ANOVA with adjustment for multiple comparisons (b, d, e, f and g). Unpaired two-sample Student’s t test (c). All the statistical test used in this figure was two-sided and two-sided P < 0.05 was considered statistically significant.

Extended Data Fig. 8 Pre-operative exercise promotes anti-inflammatory trained immunity in Kupffer cells.

a, Experiment outline. b, Trained immunity marker IL-1Ra (P = 0.0035), IL-10 (P = 0.0207), IL-1β (P = 0.0218), IL-6 (P = 0.0069) and TNF-α (P = 0.0145) were measured from the medium of Kupffer cells which isolated from normal mice with re-stimulation of LPS by ELISA (n = 3 mice per group). c, Experiment outline. d, Trained immunity marker IL-1Ra (P = 0.6551, P = 0.0229, P = 0.9025, P = 0.0032 between sedentary and exercise group at the time point of baseline, after exercise, before hypoxia, and after hypoxia) and IL-10(P = 0.5531, P = 0.0149, P = 0.4440, P = 0.0223 between sedentary and exercise group at the time point of baseline, after exercise, before hypoxia, and after hypoxia), IL-1β (P = 0.6841, P = 0.0188, P = 0.3258, P = 0.0226 between sedentary and exercise group at the time point of baseline, after exercise, before hypoxia, and after hypoxia), IL-6 (P = 0.5076, P = 0.0148, P = 0.3301, P = 0.0084 between sedentary and exercise group at the time point of baseline, after exercise, before hypoxia, and after hypoxia) and TNF-α (P = 0.7328, P = 0.0088, P = 0.8229, P = 0.0202 between sedentary and exercise group at the time point of baseline, after exercise, before hypoxia, and after hypoxia) were measured from the medium of Kupffer cells which isolated from mice received sedentary or pre-operative exercise treatment for 4 weeks before and after the second stimulation of LPS by ELISA (Each dot represents one mouse, n = 6 mice per group). e, Experiment outline. f, Liver damage was measured by serum ALT level (P < 0.0001 between sedentary and all exercise groups, p = 0.8403 between 0 days and 3 days group, P = 0.5430 between 0 day and 5 days group, P = 0.2980 between 0 day and 7 days group, P = 0.5802 between 3 days and 5 days group, P = 0.2119 between 3 days and 7 days group, P = 0.4277 between 5 days and 7 days group) and liver histological evaluation (necrotic areas) (P = 0.0482 between sedentary and 0 day after exercise group, P = 0.0061 between sedentary and 3 days after exercise group, P = 0.0424 between sedentary and 5 days after exercise group, P = 0.0260 between sedentary and 7 days after exercise group, p = 0.1908 between 0 day and 3 days group, P = 0.8088 between 0 day and 5 days group, P = 0.8722 between 0 day and 7 days group, P = 0.0599 between 3 days and 5 days group, P = 0.1512 between 3 days and 7 days group, P = 0.6227 between 5 days and 7 days group) from mice with sedentary or pre-operative exercise treatment for 4 weeks and resting for 0, 3, 5, or 7 days before LPS injection (Each dot represents one mouse, n = 11 in sedentary group, n = 7 in 0D after Exercise group, n = 8 in 3D after Exercise group, n = 9 in 5D after Exercise group, n = 7 in 7D after Exercise group). g, IL-1Ra (P < 0.0001), IL-10 (P = 0.0023), IL-1β (P = 0.0293), IL-6 (P = 0.0003) and TNF-α (P = 0.0007) concentrations were measured from the medium of Kupffer cells which isolated from mice with sedentary or pre-operative exercise treatment for 4 weeks with LPS injection (Each dot represents one mouse, n = 6 mice per group). Graphs show the mean ± SD (b, d, f and g). Circles, triangles and squares represent individual mice. Unpaired two-sample Student’s t test for (b and g). ANOVA with adjustment for multiple comparisons for (d and f). NS: not significant. All the statistical test used in this figure was two-sided and two-sided P < 0.05 was considered statistically significant.

Extended Data Fig. 9 Anti-inflammatory trained immunity in Kupffer cells relies on anti-inflammatory metabolite, itaconate.

a, Experiment outline. b, Viability of 24 hours in unstimulated Kupffer cells treated with 4-OI and Itaconate. The treatment concentrations for all experiments listed in mM. c, IL-1Ra (P < 0.0001 between Vehicle and 4-OI treatment, P = 0.0016 between Vehicle and Itaconate treatment), IL-10 (P < 0.0001 between Vehicle and 4-OI treatment, P = 0.0497 between Vehicle and Itaconate treatment), IL-1β (P = 0.0015 between Vehicle and 4-OI treatment, P = 0.0207 between Vehicle and Itaconate treatment), IL-6 (P = 0.0002 between Vehicle and 4-OI treatment, P = 0.0013 between Vehicle and Itaconate treatment) and TNF-α (P = 0.0016 between Vehicle and 4-OI treatment, P = 0.0008 between Vehicle and Itaconate treatment) concentrations from the medium of Kupffer cells which isolated from normal mice with the stimulation of vehicle or 4-OI or Itaconate after the challenge of LPS or hypoxia were measured by ELISA (Each dot represents one mouse, n = 6 mice per group). d, Experiment outline e, IL-1Ra (P = 0.0172 between Vehicle and 4-OI treatment, P = 0.0463 between Vehicle and Itaconate treatment), IL-10 (P = 0.0264 between Vehicle and 4-OI treatment, P = 0.0015 between Vehicle and Itaconate treatment), IL-1β (P = 0.0498 between Vehicle and 4-OI treatment, P = 0.0315 between Vehicle and Itaconate treatment), IL-6 (P = 0.0431 between Vehicle and 4-OI treatment, P = 0.0193 between Vehicle and Itaconate treatment) and TNF-α (P = 0.0035 between Vehicle and 4-OI treatment, P = 0.0104 between Vehicle and Itaconate treatment) concentrations from the medium of Kupffer cells which isolated from Irg1−/− l mice with the stimulation of vehicle, 4-OI or itaconate after the challenge of hypoxia were measured by ELISA (Each dot represents one mouse, n = 6 mice per group). f, IL-1Ra (P = 0.0476 between Vehicle and 4-OI treatment, P = 0.0100 between Vehicle and Itaconate treatment), IL-10 (P = 0.0114 between Vehicle and 4-OI treatment, P = 0.0487 between Vehicle and Itaconate treatment), IL-1β (P = 0.0406 between Vehicle and 4-OI treatment, P = 0.0048 between Vehicle and Itaconate treatment), IL-6 (P = 0.0278 between Vehicle and 4-OI treatment, P = 0.0094 between Vehicle and Itaconate treatment) and TNF-α (P = 0.0208 between Vehicle and 4-OI treatment, P = 0.0184 between Vehicle and Itaconate treatment) concentrations from the medium of Kupffer cells which isolated from Irg1−/− l mice with the stimulation of vehicle, 4-OI or itaconate after the challenge of LPS were measured by ELISA (Each dot represents one mouse, n = 6 mice per group). g, Experiment outline. h, IL-1Ra (P = 0.0543), IL-10 (P = 0.0623), IL-1β (P = 0.2723), IL-6 (P = 0.06754) and TNF-α (P = 0.9481) concentrations from the medium of Kupffer cells which isolated from Irg1−/− mice with sedentary or pre-operative exercise treatment after the challenge of LPS or hypoxia for 4 weeks were measured by ELISA (Each dot represents one mouse, n = 6 mice per group). Graphs show the mean ± SD (c, e, f and h). Circles represent individual mice. Unpaired two-sample Student’s t test for (c, e, f and h). All the statistical test used in this figure was two-sided and two-sided P < 0.05 was considered statistically significant.

Supplementary information

Supplementary Information

Supplementary Fig. 1

Reporting Summary

Supplementary Data 1

Custom code used for transcriptomic profiling in Fig. 2a–c and Extended Data Fig. 3b,c.

Supplementary Data 2

Custom code used for heatmap generation in Fig. 2d.

Supplementary Data 3

Custom code used for pathway analysis in Fig. 2d.

Supplementary Data 4

Custom code used for Preference of cluster analysis in Extended Data Fig. 4d.

Supplementary Data 5

Custom code used for the volcano plot analysis in Extended Data Fig. 4e.

Source data

Source Data Fig. 1

Uncropped scans of the gels: uncropped western blots images. The dashed boxes denote cropped images that are presented in the manuscript.

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Zhang, H., Chen, T., Ren, J. et al. Pre-operative exercise therapy triggers anti-inflammatory trained immunity of Kupffer cells through metabolic reprogramming. Nat Metab 3, 843–858 (2021). https://doi.org/10.1038/s42255-021-00402-x

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