interleukin 10  (R&D Systems)

Journal: Bioactive Materials

Article Title: M2 macrophage-derived exosomes delivering haptoglobin and interleukin-10 plasmids for synergistic therapy of intracerebral hemorrhage

doi: 10.1016/j.bioactmat.2026.01.047

Figure Lengend Snippet: Schematic illustration of the preparation of M2-exo@HI and its mediated therapeutic mechanisms and signaling pathways. (a) RAW264.7 macrophages were polarized to M2 phenotype using DEX, followed by M2-exo isolation from cell supernatant via differential centrifugation. M2-exo@HI was prepared through encapsulating HI into M2-exo by electroporation. (b) M2-exo@HI crossed the BBB and localized to microglia in the hemorrhagic brain, delivering the HI plasmid into the nucleus. This prompted expression and secretion of Hp and IL-10 by microglia. The released Hp inhibited Hb toxicity by binding to Hb. IL-10 shifted microglial polarization from the pro-inflammatory M1 phenotype toward the reparative M2 phenotype, removing hematoma by engulfing erythrocyte and Hb-Hp complex, and decreasing pro-inflammatory cytokine levels—collectively enhancing neuroprotection and BBB repair. These beneficial outcomes were linked to the inhibition of the IL-17 and NF-κB signaling pathways and activation of the PI3K-Akt and ferroptosis pathways.

Article Snippet: ELISA kits included mouse TNF- α ELISA kit (Solarbio), mouse IL-10 ELISA kit (Elabscience), mouse Hp ELISA kit (Elabscience), mouse IL-1β ELISA kit (Solarbio), and TGF-β ELISA kit (Solarbio).

Techniques: Protein-Protein interactions, Isolation, Centrifugation, Electroporation, Plasmid Preparation, Expressing, Binding Assay, Inhibition, Activation Assay

Journal: Bioactive Materials

Article Title: M2 macrophage-derived exosomes delivering haptoglobin and interleukin-10 plasmids for synergistic therapy of intracerebral hemorrhage

doi: 10.1016/j.bioactmat.2026.01.047

Figure Lengend Snippet: Characterization of M2-exo@HI. (a) Western blot of Tsg101, CD9, and Calnexin expressions in RAW264.7 and M2-exo. (b) Protein bands of RAW264.7, M2-exo, M2-exo@HI, and HI by SDS-PAGE. (c) Fluorescence microscopy images showing Hp-EGFP and IL-10-mCherry expression in 293T cells infected with lipo2000@pBudCE4.1 , lipo2000@HI, and M2-exo@HI respectively, after 24 h. (d,e) Representative TEM images and size distribution profiles of M2-exo and M2-exo@HI. (f) Particle number of M2-exo and M2-exo@HI by NTA measurement. (g) Zeta potentials of M2-exo and M2-exo@HI (n = 3). (h) Drug release profiles of M2-exo@HI at pH 6.5 and pH 7.4, respectively (n = 3). (i) Stability evaluation of M2-exo@HI in PBS at 4 °C by monitoring particle size over time (n = 3). Data are presented as mean ± SD.

Article Snippet: ELISA kits included mouse TNF- α ELISA kit (Solarbio), mouse IL-10 ELISA kit (Elabscience), mouse Hp ELISA kit (Elabscience), mouse IL-1β ELISA kit (Solarbio), and TGF-β ELISA kit (Solarbio).

Techniques: Western Blot, SDS Page, Fluorescence, Microscopy, Expressing, Infection

Journal: Bioactive Materials

Article Title: M2 macrophage-derived exosomes delivering haptoglobin and interleukin-10 plasmids for synergistic therapy of intracerebral hemorrhage

doi: 10.1016/j.bioactmat.2026.01.047

Figure Lengend Snippet: Validation of M2-exo targeting, Hp/IL-10 transfection expression, and Hp/Hb binding. (a) Fluorescence imaging showing cellular uptake of ICG and M2-exo@ICG by M1 microglia. (b,c) Flow cytometry and corresponding quantification of RhB and M2-exo@RhB internalized by M1 microglia (n = 3). (d,e) Schematic illustration and quantitative analysis of the in vitro phagocytosis-release kinetics of M2-exo@RhB in BV2 under ICH-mimicking stimulation (n = 6). (f) Fluorescence images showing Hp and IL-10 expression in M1 microglia treated with M2-exo@HI for 12, 24, 48, 72 h. (g) Mean fluorescence intensity (MFI) quantification of Hp and IL-10 expression (n = 3). (h,i) ELISA measurements of secreted Hp and IL-10 protein levels (n = 3). (j,k) qPCR analysis of relative Hp and IL-10 mRNA expression (n = 3). (l) Western blot detection of Hp and IL-10 protein expression. (m) Densitometric quantification of Hp and IL-10 protein levels from Western blot (n = 3). (n) Co-immunoprecipitation assay confirming the formation of Hp-Hb complex. Data are presented as mean ± SD. Statistical significance was calculated by unpaired Student's t -test (c and e), and one-way ANOVA with Tukey's multiple comparisons test (g-k and m).

Article Snippet: ELISA kits included mouse TNF- α ELISA kit (Solarbio), mouse IL-10 ELISA kit (Elabscience), mouse Hp ELISA kit (Elabscience), mouse IL-1β ELISA kit (Solarbio), and TGF-β ELISA kit (Solarbio).

Techniques: Biomarker Discovery, Transfection, Expressing, Binding Assay, Fluorescence, Imaging, Flow Cytometry, In Vitro, Enzyme-linked Immunosorbent Assay, Western Blot, Co-Immunoprecipitation Assay

Journal: Bioactive Materials

Article Title: M2 macrophage-derived exosomes delivering haptoglobin and interleukin-10 plasmids for synergistic therapy of intracerebral hemorrhage

doi: 10.1016/j.bioactmat.2026.01.047

Figure Lengend Snippet: M2-exo@HI promotes in vitro microglia polarization, BBB repair and neuroprotection. (a) Flow cytometry analysis of M1-type (CD86 + ) and M2-type microglia (CD163 + ) following treatment with different formulations. (b,c) Percentages of CD86 + and CD163 + microglia populations (n = 3). (d–g) The cytokine levels of IL-10, TGF-β, TNF-α, and IL-1β in treated microglia (n = 3). (h) Fluorescence microscopy images showing erythrophagocytosis by microglia across treatment groups. (i) Schematic of the in vitro BBB model assessing FITC-dextran permeability using a transwell assay. (j) Quantitative analysis of FITC-dextran penetration (n = 7). (k) Flow cytometry analysis of neuronal apoptosis across treatments (n = 3). (l) Quantitative analysis of neuronal apoptosis (n = 3). Data are presented as mean ± SD. Statistical significance was tested by one-way ANOVA with Tukey's multiple comparisons test.

Article Snippet: ELISA kits included mouse TNF- α ELISA kit (Solarbio), mouse IL-10 ELISA kit (Elabscience), mouse Hp ELISA kit (Elabscience), mouse IL-1β ELISA kit (Solarbio), and TGF-β ELISA kit (Solarbio).

Techniques: In Vitro, Flow Cytometry, Fluorescence, Microscopy, Permeability, Transwell Assay

Journal: Bioactive Materials

Article Title: M2 macrophage-derived exosomes delivering haptoglobin and interleukin-10 plasmids for synergistic therapy of intracerebral hemorrhage

doi: 10.1016/j.bioactmat.2026.01.047

Figure Lengend Snippet: Targeted delivery and therapeutic gene expression of M2-exo@HI in hemorrhagic brain. (a) In vivo near-infrared fluorescence imaging showing ICG and M2-exo@ICG in mouse brains at various time points post-injection. (b) Average radiation efficiency of ICG in different treatment groups (n = 3). (c) Ex vivo fluorescence imaging of major organs harvested 24 h post-injection. (d) Average radiation efficiency of ICG in different in mouse tissues (n = 3). (e) Time-dependent accumulation of M2-exo@ICG at hematoma regions. (f) Immunofluorescence staining showing co-localization of Hp/IL-10 with astrocytes, microglia, and endothelial cells. (g) Temporal expression profiles of Hp and IL-10 in brain tissues. (h,i) ELISA quantification of Hp and IL-10 protein levels in brain homogenates (n = 3). Data are presented as mean ± SD. Statistical significance was calculated by unpaired Student's t -test (b,d), and one-way ANOVA with Tukey's multiple comparisons test (h,i).

Article Snippet: ELISA kits included mouse TNF- α ELISA kit (Solarbio), mouse IL-10 ELISA kit (Elabscience), mouse Hp ELISA kit (Elabscience), mouse IL-1β ELISA kit (Solarbio), and TGF-β ELISA kit (Solarbio).

Techniques: Gene Expression, In Vivo, Fluorescence, Imaging, Injection, Ex Vivo, Immunofluorescence, Staining, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Journal of Inflammation Research

Article Title: Acupuncture Activates PI3K/Akt Pathway to Promote Spinal Microglial M2 Polarization and Alleviate Spastic Hemiplegia Following Ischemic Stroke

doi: 10.2147/JIR.S592917

Figure Lengend Snippet: Acupuncture shifted spinal microglia toward the M2 phenotype, modified spinal inflammation milieu, restored the E/I balancing in the spinal motor circuit and activated the spinal PI3K/Akt pathway in MCAO rats with spastic hemiplegia. Immunofluorescence of microglial polarization state: representative images of CD32 (M1 marker, green) and Iba-1 (microglial marker, red) co-localization by immunofluorescence ( A ); representative images of CD206 (M2 marker, green) and Iba-1 (red) co-localization by immunofluorescence ( B ) with all nuclei of cells stained by DAPI (blue); scale bar, 50 μm; the semi-quantitative fluorescent intensity results of CD32 ( C ) and CD206 ( D ). Immunofluorescence of synaptic boutons in motor circuit: representative images of vGluT1 immunoreactive boutons (marking the proprioceptive terminals, red) on CTB-labeled motor neurons (MNs, green) ( E ); representative images of vGluT1 boutons (red) contacting vGAT immunoreactive boutons (marking GABAergic synapses, green) ( F ); scale bar, 10 μm, 2 μm; fold change of vGluT1 boutons on MNs ( G ); ratio of vGluT1 boutons opposed by vGAT boutons ( H ). Representative Western blot bands ( I ) and quantitative results for CD32 ( J ), CD206 ( K ), vGluT1 ( L ), vGAT ( M ), p-PI3K/PI3K ( N ), and p-Akt/Akt ( O ) protein expressions relative to GAPDH protein. The mRNA levels of CD32 ( P ) and CD206 ( Q ) were detected by RT-qPCR. The concentrations of TNF-α ( R ), IL-6 ( S ), TGF-β ( T ), IL-10 ( U ), Glu ( V ), GABA ( W ) were detected by ELISA. N = 6 per group. Data were expressed as mean ± SD. **P<0.01,***P<0.001 versus M+AP group; ### P<0.001 versus M group.

Article Snippet: The levels of TNF-α (EK0526, Boster Biological), IL-6 (EK0412, Boster Biological), and TGF-β (EK0514, Boster Biological), IL-10 (EK0418, Boster Biological) were determined to assess neuroinflammation.

Techniques: Modification, Immunofluorescence, Marker, Staining, Labeling, Western Blot, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Journal of Inflammation Research

Article Title: Acupuncture Activates PI3K/Akt Pathway to Promote Spinal Microglial M2 Polarization and Alleviate Spastic Hemiplegia Following Ischemic Stroke

doi: 10.2147/JIR.S592917

Figure Lengend Snippet: Inhibition of M1 polarization of microglia mitigated spinal inflammatory milieu and facilitated the restoration of spinal E/I balance in MCAO rats with spastic hemiplegia. Immunofluorescence of microglial polarization state: representative images of CD32 (green) and Iba-1 (red) co-localization by immunofluorescence ( A ) representative images of CD206 (green) and Iba-1 (red) co-localization by immunofluorescence ( B ) with all nuclei of cells stained by DAPI (blue); scale bar, 50 μm; the semi-quantitative fluorescent intensity results of CD32 ( C ) and CD206 ( D ). Immunofluorescence of synaptic boutons in motor circuit: representative images of vGluT1 immunoreactive boutons (red) on CTB-labeled MNs (green) ( E ) representative images of vGluT1 boutons (red) contacting vGAT immunoreactive boutons (green) ( F ) scale bar, 10 μm, 2 μm; fold change of vGluT1 boutons on MNs ( G ) ratio of vGluT1 boutons opposed by vGAT boutons ( H ). Representative Western blot bands ( I ) and quantitative results for CD32 ( J ) CD206 ( K ) vGluT1 ( L ) vGAT ( M ) protein expression relative to GAPDH protein. The mRNA levels of CD32 ( N ) and CD206 ( O ) were detected by RT-qPCR. The concentrations of TNF-α ( P ) IL-6 ( Q ) TGF-β ( R ) IL-10 ( S ) Glu ( T ) GABA ( U ) were detected by ELISA. N = 6 per group. Data were expressed as mean ± SD. **P<0.01, ***P<0.001 versus M+MC group; ### P<0.001 versus M group.

Article Snippet: The levels of TNF-α (EK0526, Boster Biological), IL-6 (EK0412, Boster Biological), and TGF-β (EK0514, Boster Biological), IL-10 (EK0418, Boster Biological) were determined to assess neuroinflammation.

Techniques: Inhibition, Immunofluorescence, Staining, Labeling, Western Blot, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Journal of Inflammation Research

Article Title: Acupuncture Activates PI3K/Akt Pathway to Promote Spinal Microglial M2 Polarization and Alleviate Spastic Hemiplegia Following Ischemic Stroke

doi: 10.2147/JIR.S592917

Figure Lengend Snippet: Acupuncture mitigated excitability of spinal motor circuits and inflammatory milieu via PI3K/Akt pathway in MCAO rats with spastic hemiplegia. Immunofluorescence of synaptic boutons in motor circuit: representative images of vGluT1 immunoreactive boutons (red) on CTB-labeled MNs (green) ( A ); representative images of vGluT1 boutons (red) contacting vGAT immunoreactive boutons (green) ( B ); scale bar, 10 μm, 2 μm; fold change of vGluT1 boutons on MNs ( C ); ratio of vGluT1 boutons opposed by vGAT boutons ( D ). Representative Western blot bands ( E ) and quantitative results for CD32 ( F ) CD206 ( G ) vGluT1 ( H ) vGAT ( I ) protein expression relative to GAPDH protein. The mRNA levels of CD32 ( J ) and CD206 ( K ) were detected by RT-qPCR. The concentrations of TNF-α ( L ) IL-6 ( M ) TGF-β ( N ) IL-10 ( O ) Glu ( P ) GABA ( Q ) were detected by ELISA. N = 6 per group. Data were expressed as mean ± SD. ***P<0.001 versus M+AP+Veh group; ## P<0.01, ### P<0.001 versus M+Veh group.

Article Snippet: The levels of TNF-α (EK0526, Boster Biological), IL-6 (EK0412, Boster Biological), and TGF-β (EK0514, Boster Biological), IL-10 (EK0418, Boster Biological) were determined to assess neuroinflammation.

Techniques: Immunofluorescence, Labeling, Western Blot, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay