il 13  (R&D Systems)

Journal: iScience

Article Title: 3-Hydroxypropionic acid converts inflammatory macrophage glycolysis into mitochondrial oxidation through GAPDH carboxyethylation

doi: 10.1016/j.isci.2026.116258

Figure Lengend Snippet: 3-HPA inhibits the secretion of inflammatory factors and glycolysis in macrophages (A and B) Schematic diagram of THP-1 cell (A) and BMDMs (B) activation into pro-inflammatory macrophages. Created with BioRender.com. (C) The concentrations of IL-6, TNF-α, and IL-1β in THP-1 cell supernatants were quantified by ELISA after 24 h treatment with LPS (100 ng/mL), or LPS+3-HPA (5 mM). Data are the means ± SD and n = 3 per group. Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test with ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001. (D) The concentrations of IL-6, TNF-α, and IL-1β in supernatants of THP-1 cells treated with different concentrations of 3-HPA (0, 0.625, 1.25, 5 mM) followed by LPS stimulation. Data are the means ± SD and n = 3 per group. Statistical significance was determined using one-way ANOVA followed by Dunnett’s multiple comparisons test ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001. (E) The concentrations of IL-6, TNF-α, and IL-1β in BMDM cell supernatants were quantified by ELISA after 24 h treatment with LPS (100 ng/mL) or LPS+3-HPA (5 mM). Data are the means ± SD and n = 3 per group. Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test with ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001. (F) Bubble plot of KEGG pathway enrichment analysis for differentially expressed genes between LPS (100 ng/mL) and LPS+3-HPA (5 mM) treated in THP-1 cells. The size of each bubble represents the number of differentially expressed genes, and the color indicates the enrichment factor. (G and H) Pyruvate and lactate levels in THP-1 cells (G) and BMDMs (H) treated with LPS (100 ng/mL), or LPS+3-HPA (5 mM). (I) Immunoblots of protein expression levels of HK, GAPDH, PKM, and LDHA in THP-1 cells treated with LPS (100 ng/mL), or LPS+3-HPA (5 mM), and quantitative results of GAPDH. (J) The mRNA levels of GAPDH in THP-1 cells treated with LPS (100 ng/mL) or LPS+3-HPA (5 mM). (K) GAPDH activity assay in THP-1 cells and BMDMs treated with PBS, LPS (100 ng/mL), or LPS+3-HPA (5 mM). Data in (G–K) are the means ± SD and n = 3 per group. Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test with ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001; ns, not significant.

Article Snippet: Mouse IL-6 Precoated ELISA Kit , Dakewe , 1210602.

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Activity Assay

Journal: iScience

Article Title: 3-Hydroxypropionic acid converts inflammatory macrophage glycolysis into mitochondrial oxidation through GAPDH carboxyethylation

doi: 10.1016/j.isci.2026.116258

Figure Lengend Snippet: 3-HPA enhances the metabolite content of the TCA cycle and mitochondrial oxidation (A) Schematic diagram of THP-1 with 2-DG treatment. Created with BioRender.com. (B) Concentrations of IL-6, TNF-α, and IL-1β in supernatants of THP-1 cells treated with LPS, LPS+3-HPA, LPS+2-DG, or LPS+3-HPA+2-DG. Data are the means ± SD and n = 3 per group. Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test with ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001; ns, not significant. (C) Schematic diagram of THP-1 with high glucose treatment. Created with BioRender.com. (D) Concentrations of IL-6, TNF-α, and IL-1β in supernatants of THP-1 cells treated with LPS+3-HPA, LPS+3-HPA+glucose. Data are the means ± SD and n = 3 per group. Statistical significance was determined using unpaired Student’s t test with ∗∗∗p < 0.001; ns, not significant. (E) KEGG pathway enrichment analysis of metabolic pathways in BMDM cells treated with LPS or LPS+3-HPA. (F) Relative abundance of metabolite (ornithine, citrulline, L-malate, succinic acid, trans-aconitic acid, cis-aconitic acid) in BMDM cells treated with LPS or LPS+3-HPA. Data are the means ± SD and n = 4 per group. Statistical significance was determined using unpaired Student’s t test with ∗p < 0.05; ∗∗p < 0.01. (G) Correlation network of metabolites and genes in the metabolic pathway. Nodes represent metabolites (blue squares) and genes (colored circles). Gray edges indicate pairwise correlations between metabolites and genes. The colors similarly represent expression levels, with red typically indicating higher expression and blue indicating lower expression compared to the mean. (H) Schematic diagram of arginine metabolism and the TCA cycle. (I) Heatmap of mitochondrial oxidation-related gene expression associated with differentially expressed metabolites. Red indicates relatively high gene expression, while blue indicates relatively low gene expression within each row. (J) Schematic diagram of the catalytic function of the GAPDH enzyme. (K) Concentrations of the NAD + /NADH ratio in THP-1 cells and BMDM cells treated with PBS, LPS, or LPS+3-HPA. Data are the means ± SD and n = 4 per group. Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test with ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗∗ p < 0.0001; (L) ATP concentrations in THP-1 cells and BMDMs treated with PBS, LPS, or LPS+3-HPA. Data are the means ± SD and n = 3 per group. Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test with ∗ p < 0.05; ∗∗∗∗ p < 0.0001; ns, not significant. (M) Representative images and mitochondrial analysis of BMDM cells treated with PBS, LPS, or LPS+3-HPA. Scale bars, 1 μm (upper) and 0.25 μm (lower). For mitochondrial number, n = 8–12 ( n = 12 for PBS, n = 8 for LPS, n = 9 for LPS+3-HPA group).

Article Snippet: Mouse IL-6 Precoated ELISA Kit , Dakewe , 1210602.

Techniques: Expressing, Gene Expression

Journal: iScience

Article Title: 3-Hydroxypropionic acid converts inflammatory macrophage glycolysis into mitochondrial oxidation through GAPDH carboxyethylation

doi: 10.1016/j.isci.2026.116258

Figure Lengend Snippet: GAPDH carboxyethylation inhibited macrophage glycolysis and the release of inflammatory factors (A) Schematic workflow illustrating the strategy of silencing endogenous GAPDH via 3′UTR-targeting siRNA and overexpressing exogenous GAPDH. (B) Immunoblot and quantitative analysis of GAPDH protein in 293 T cells transfected with GAPDH 3′UTR-targeting siRNAs (siGAPDH 1, siGAPDH 2) or siRNA NC. Data are the means ± SD and n = 3 per group. Statistical significance was determined using one-way ANOVA followed by Dunnett’s multiple comparisons test ∗∗p < 0.01. (C) Relative mRNA expression of GAPDH in 293 T cells transfected with GAPDH 3′UTR-targeting siRNAs (siGAPDH 1, siGAPDH 2) or siRNA NC. Data are the means ± SD and n = 3 per group. Statistical significance was determined using one-way ANOVA followed by Dunnett’s multiple comparisons test ∗∗∗∗p < 0.0001. (D) Immunoblot analysis of FLAG-tagged exogenous GAPDH(E) and GAPDH in 293 T cells. Knockdown of endogenous GAPDH with siRNA followed by the overexpression of GAPDH (E). Data are the means ± SD and n = 3 per group. Statistical significance was determined using one-way ANOVA followed by Dunnett’s multiple comparisons test with ∗p < 0.05; ns, not significant. (E) Relative mRNA expression of GAPDH in 293 T cells knockdowned endogenous GAPDH (siGAPDH) and overexpressed GAPDH (C) and GAPDH (E), respectively. Data are the means ± SD and n = 3 per group. Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test with ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant. (F) GAPDH activity assay in 293 T cells transfected with GAPDH 3′UTR siRNA, and overexpressing GAPDH(C), GAPDH(E). Data are the means ± SD and n = 4 per group. Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test with ∗p < 0.05; ∗∗∗∗p < 0.0001; ns, not significant. (G) Concentrations of lactate and pyruvate in 293 T cells transfected with GAPDH 3′UTR siRNA, and overexpressing GAPDH(C), GAPDH(E). Data are the means ± SD and n = 4 per group. Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test with ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗∗p < 0.0001. (H) Relative mRNA expression of IL-6 , TNF-α , and IL-1β in THP-1 cells which transfected with GAPDH 3′UTR siRNA, and overexpressing GAPDH(C) or GAPDH(E). Data are the means ± SD and n = 3 per group. Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test with ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, not significant. (I) The concentration of TNF-α in THP-1 cells that overexpressed GAPDH(C) or GAPDH(E). Data are the means ± SD and n = 3 per group. Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test with ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001; ns, not significant.

Article Snippet: Mouse IL-6 Precoated ELISA Kit , Dakewe , 1210602.

Techniques: Western Blot, Transfection, Expressing, Knockdown, Over Expression, Activity Assay, Concentration Assay

Journal: Redox Biology

Article Title: Nrf2 pathway mediates copper oxide nanoparticle-induced exacerbation of allergic asthma

doi: 10.1016/j.redox.2026.104180

Figure Lengend Snippet: Effects of CuONPs on allergic inflammation and oxidative stress in OVA-induced asthmatic mice. (A) Airway hyperresponsiveness assessed as total respiratory system resistance in response to methacholine challenge (10, 20, and 40 mg/mL). (B–F) Total and differential inflammatory cell counts in BALF. (G–L) IL-1β, IL-6, TNF-α, IL-4, IL-5, and IL-13 levels in BALF, measured by ELISA. (M and N) Total IgE and OVA specific IgE levels in serum. (O and P) MDA levels and SOD activity in lung tissue. Data are presented as means ± SD (n = 6 mice/group). ∗∗ p < 0.01 and ∗∗∗ p < 0.001 versus NC group. # p < 0.05, ## p < 0.01, and ### p < 0.001 versus OVA group.

Article Snippet: BALF was centrifuged at 300× g for 10 min at 4 °C, and the supernatant was stored for cytokine analysis using commercially available enzyme-linked immunosorbent assay (ELISA) kits to quantify IL-1β, IL-6, tumor necrosis factor (TNF)-α, IL-4, IL-5, and IL-13 (R&D Systems, Minneapolis, MN, USA; Cat. No. MLB00C, M6000B, MTA00B, M4000B, M5000, and M1300CB, respectively).

Techniques: Enzyme-linked Immunosorbent Assay, Activity Assay

Journal: Redox Biology

Article Title: Nrf2 pathway mediates copper oxide nanoparticle-induced exacerbation of allergic asthma

doi: 10.1016/j.redox.2026.104180

Figure Lengend Snippet: Effects of Nrf2 overexpression on allergic inflammation and oxidative stress in CuONP-exposed asthmatic mice. (A) Immunofluorescence analysis of lung tissue from mice administered PBS or AAV2/8-GFP via intratracheal instillation. (B) Airway hyperresponsiveness assessed as total respiratory system resistance in response to methacholine challenge (10, 20, and 40 mg/mL). (C–G) Total and differential inflammatory cell counts in BALF. (H–M) IL-1β, IL-6, TNF-α, IL-4, IL-5, and IL-13 levels in BALF, measured by ELISA. (N and O) Total IgE and OVA specific IgE levels in serum. (P and Q) MDA levels and SOD activity in lung tissue. Data are presented as means ± SD (n = 3 mice/group for panels A; n = 6 mice/group for panels B–Q). In panel B, ## p < 0.01 indicates significant differences between GFP-NC and GFP-OVA, and ∗∗ p < 0.01 indicates significant differences between GFP-OVA + CuONPs and Nrf2-OVA + CuONPs. For panels C–Q, ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 indicate significant differences between AAV-GFP and AAV-Nrf2 within each condition.

Article Snippet: BALF was centrifuged at 300× g for 10 min at 4 °C, and the supernatant was stored for cytokine analysis using commercially available enzyme-linked immunosorbent assay (ELISA) kits to quantify IL-1β, IL-6, tumor necrosis factor (TNF)-α, IL-4, IL-5, and IL-13 (R&D Systems, Minneapolis, MN, USA; Cat. No. MLB00C, M6000B, MTA00B, M4000B, M5000, and M1300CB, respectively).

Techniques: Over Expression, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Activity Assay

Journal: Materials Today Bio

Article Title: Prodrug-integrated multifunctional liposome encapsulation enhances probiotic-based bacteriotherapy for inflammatory bowel disease

doi: 10.1016/j.mtbio.2026.103110

Figure Lengend Snippet: Preventative efficacy of DTI@EcN against DSS-induced colitis. ( A ) Schematic of the experimental schedule. Mice received daily oral gavage of various bacterial formulations or drugs throughout the experimental period (bacteria: 5 × 10 8 CFU; IDE: 65 mg/kg). 3% DSS was given via drinking water from day 4 to day 8 to induce colitis. ( B ) Quantitative assessment of body weight variation in colitis mice across treatment groups. ( C ) Macroscopic appearance of colon tissues and ( D ) corresponding length measurements. ( E ) Histopathological evaluation by H&E staining and ( F ) histopathological scores of colon tissues based on H&E images. Scale bar: 100 μm. ( G ) Immunofluorescence localization of tight junction proteins ZO-1 (red) and Occludin (green) in colonic epithelia. Cell nuclei were stained with DAPI (blue). Scale bar: 100 μm. ( H ) Quantified ZO-1 and ( I ) occludin fluorescence intensities in colon tissues shown in (G) through ImageJ. ( J ) LSCM images showing DCF fluorescence in colon tissues incubated with DCFH-DA, representing the relative ROS levels across different treatment groups. Scale bar: 100 μm. ( K ) Quantified DCF fluorescence intensities from (J), analyzed with ImageJ. ( L ) Relative colonic MPO activities in colon tissues in different treatment groups. ( M - P ) Levels of pro-inflammatory cytokines, including IL-6 ( M ), IL-1β ( N ), TNF-α (O), and IFN-γ ( P ), in colon tissues, determined by ELISA kits. Data represent the mean ± SEM (n = 5). Statistical analysis was performed using one-way ANOVA. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The selected chemicals and biological materials used in this research are as follows: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI, Macklin), 4-bromobutyric acid (Bidepharm), thiourea (Macklin), sodium hydroxide (NaOH, Macklin), hydrochloric acid (HCl, Macklin), trifluoroacetic acid (Bidepharm), idebenone (Bidepharm), hexadecyltrimethylammonium bromide (Macklin), 4-dimethylaminopyridine (DMAP, Macklin), N-hydroxysuccinimide (NHS, Macklin), 2′,7′-dichlorofluorescin diacetate (DCFH-DA, Aladdin), DSPE-PEG2000-NH 2 (Yusi), DSPE-PEG2000-FITC (Yusi), cholesterol (Yuanye), trypsin (Macklin), pepsin powder (Bidepharm), bile salts (Macklin), ampicillin (Bidepharm), lysin (Bidepharm), dextran sulfate sodium salt (DSS, MW 40,000, MP Bio), LB nutrient agar (Hopebiol), Cell Counting Kit-8 (CCK-8, Beyotime), mouse IL-6, IFN-γ, IL-1β, and TNF-α uncoated ELISA kits (Thermo Fisher Scientific), and antigen retrieval solution (Servicebio).

Techniques: Bacteria, Staining, Immunofluorescence, Fluorescence, Incubation, Enzyme-linked Immunosorbent Assay

Journal: Materials Today Bio

Article Title: Prodrug-integrated multifunctional liposome encapsulation enhances probiotic-based bacteriotherapy for inflammatory bowel disease

doi: 10.1016/j.mtbio.2026.103110

Figure Lengend Snippet: Therapeutic efficacy of DTI@EcN against DSS-induced colitis. ( A ) Schematic of the experimental schedule. Colitis was induced by administering 3% DSS in drinking water for the first 5 days. Subsequently, different treatments were administered daily for 5 days to evaluate their therapeutic efficacy. ( B ) Body weight dynamics of colitis mice under different treatments. ( C ) Macroscopic colon appearance and ( D ) corresponding length measurements across groups. ( E) Histopathological assessment by H&E staining. Scale bar: 100 μm. ( F ) Histopathology scores of colon tissues based on the H&E images. ( G ) Localization of tight junction proteins ZO-1 (red) and occludin (green) in colonic epithelium across treatment groups. Cell nuclei were stained with DAPI (blue). Scale bar: 100 μm. ( H ) Quantification of ZO-1 and ( I ) Occludin fluorescence intensities from (G), analyzed using ImageJ. ( J ) LSCM images showing DCF fluorescence in colon tissues incubated with DCFH-DA, representing the relative ROS levels across different treatment groups. Scale bar: 100 μm. ( K ) Quantitative analysis of DCF fluorescence intensities from (J). ( L ) Relative MPO activity in colon tissues across various treatment groups. ( M - P ) Levels of pro-inflammatory cytokines, including IL-6 ( M ), IL-1β ( N ), TNF-α ( O ), and IFN-γ ( P ), in colon tissues, measured by ELISA kits. Data represented the mean ± SEM (n = 5 or 6). Statistical analysis was performed using one-way ANOVA. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The selected chemicals and biological materials used in this research are as follows: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI, Macklin), 4-bromobutyric acid (Bidepharm), thiourea (Macklin), sodium hydroxide (NaOH, Macklin), hydrochloric acid (HCl, Macklin), trifluoroacetic acid (Bidepharm), idebenone (Bidepharm), hexadecyltrimethylammonium bromide (Macklin), 4-dimethylaminopyridine (DMAP, Macklin), N-hydroxysuccinimide (NHS, Macklin), 2′,7′-dichlorofluorescin diacetate (DCFH-DA, Aladdin), DSPE-PEG2000-NH 2 (Yusi), DSPE-PEG2000-FITC (Yusi), cholesterol (Yuanye), trypsin (Macklin), pepsin powder (Bidepharm), bile salts (Macklin), ampicillin (Bidepharm), lysin (Bidepharm), dextran sulfate sodium salt (DSS, MW 40,000, MP Bio), LB nutrient agar (Hopebiol), Cell Counting Kit-8 (CCK-8, Beyotime), mouse IL-6, IFN-γ, IL-1β, and TNF-α uncoated ELISA kits (Thermo Fisher Scientific), and antigen retrieval solution (Servicebio).

Techniques: Drug discovery, Staining, Histopathology, Fluorescence, Incubation, Activity Assay, Enzyme-linked Immunosorbent Assay

Journal: Materials Today Bio

Article Title: Prodrug-integrated multifunctional liposome encapsulation enhances probiotic-based bacteriotherapy for inflammatory bowel disease

doi: 10.1016/j.mtbio.2026.103110

Figure Lengend Snippet: Therapeutic efficacy of DTI@EcN against STm-induced colitis. ( A ) Schematic of the experimental schedule. C57BL/6J mice were pretreated with streptomycin (100 μL, 200 mg/mL) one day prior to oral administration with STm. Afterward, different bacterial formulations were given to mice for 4 days to evaluate their effectiveness against STm-induced colitis. ( B ) Changes in STm counts in feces during the treatment period. ( C ) Quantification of STm per mg of feces on day 4 post-administration of various EcN formulations. ( D ) STm and ( E ) EcN counts in the cecum after different treatments. ( F ) Histological evaluation of colon damage with H&E staining (Scale bar: 100 μm) and ( G ) corresponding histopathology scores, respectively. ( H ) Neutrophil infiltration measured by colonic MPO activity. ( I - L ) Levels of pro-inflammatory cytokines in colon tissues, including IL-6 ( I ), IL-1β ( J ), TNF-α ( K ), and IFN-γ ( L ), as determined by ELISA kits. Data are presented as means ± SEM (n = 5). Statistical analysis was performed using one-way ANOVA. ns: no significance, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: The selected chemicals and biological materials used in this research are as follows: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI, Macklin), 4-bromobutyric acid (Bidepharm), thiourea (Macklin), sodium hydroxide (NaOH, Macklin), hydrochloric acid (HCl, Macklin), trifluoroacetic acid (Bidepharm), idebenone (Bidepharm), hexadecyltrimethylammonium bromide (Macklin), 4-dimethylaminopyridine (DMAP, Macklin), N-hydroxysuccinimide (NHS, Macklin), 2′,7′-dichlorofluorescin diacetate (DCFH-DA, Aladdin), DSPE-PEG2000-NH 2 (Yusi), DSPE-PEG2000-FITC (Yusi), cholesterol (Yuanye), trypsin (Macklin), pepsin powder (Bidepharm), bile salts (Macklin), ampicillin (Bidepharm), lysin (Bidepharm), dextran sulfate sodium salt (DSS, MW 40,000, MP Bio), LB nutrient agar (Hopebiol), Cell Counting Kit-8 (CCK-8, Beyotime), mouse IL-6, IFN-γ, IL-1β, and TNF-α uncoated ELISA kits (Thermo Fisher Scientific), and antigen retrieval solution (Servicebio).

Techniques: Drug discovery, Staining, Histopathology, Activity Assay, Enzyme-linked Immunosorbent Assay