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HCP alleviates oxidative stress and systemic inflammatory response in HIRI. ( A ) Malondialdehyde (MDA) content in liver tissues at 24 h reperfusion ( n = 8). ( B ) Superoxide dismutase (SOD) activity in liver tissues at 24 h reperfusion ( n = 8). ( C , D ) Serum levels of tumor necrosis factor-α (TNF-α) and interleukin-6 <t>(IL-6)</t> at 24 h reperfusion ( n = 8). ( E ) Western blot analysis of inflammatory cytokine protein expression (IL-6, TNF-α, IL-1β) in liver tissues, with GAPDH as loading control (representative of three independent experiments). Data are mean ± SD. Statistical significance was determined by one-way ANOVA with Tukey’s post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. Vehicle group.
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HCP alleviates oxidative stress and systemic inflammatory response in HIRI. ( A ) Malondialdehyde (MDA) content in liver tissues at 24 h reperfusion ( n = 8). ( B ) Superoxide dismutase (SOD) activity in liver tissues at 24 h reperfusion ( n = 8). ( C , D ) Serum levels of tumor necrosis factor-α (TNF-α) and interleukin-6 <t>(IL-6)</t> at 24 h reperfusion ( n = 8). ( E ) Western blot analysis of inflammatory cytokine protein expression (IL-6, TNF-α, IL-1β) in liver tissues, with GAPDH as loading control (representative of three independent experiments). Data are mean ± SD. Statistical significance was determined by one-way ANOVA with Tukey’s post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. Vehicle group.
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In vivo study in anti-tumor immune response by analysizing the maturation, activation and infiltration of immune-related cells, as well as the level of immune-related cytokines. (A) The maturation of DCs in lymph nodes of mice; (B) The activation of CD4 + /CD8 + T cells in spleens in of mice; (C) The activation of NK cells from spleens of mice. (D–G) The quantitative analysis of flow cytometry, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 and ∗∗∗∗ p < 0.0001. (H) Immunofluorescence photos of CD4 + and CD8 + T cell infiltrating tumor tissues of mice, bar: 100 μm. (I–K) The quantitative analysis of immune-related cytokines: IL-6, TNF-α and IFN-γ, ∗ p < 0.5, ∗∗ p < 0.01, ∗∗∗ p < 0.001 and ∗∗∗∗ p < 0.0001.

Journal: Materials Today Bio

Article Title: Immediate tumor killing and long-term anti-tumor immunoreaction induced by Bufalin-loaded phototherapeutic Janus membrane in CRC postoperative therapy

doi: 10.1016/j.mtbio.2026.102824

Figure Lengend Snippet: In vivo study in anti-tumor immune response by analysizing the maturation, activation and infiltration of immune-related cells, as well as the level of immune-related cytokines. (A) The maturation of DCs in lymph nodes of mice; (B) The activation of CD4 + /CD8 + T cells in spleens in of mice; (C) The activation of NK cells from spleens of mice. (D–G) The quantitative analysis of flow cytometry, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 and ∗∗∗∗ p < 0.0001. (H) Immunofluorescence photos of CD4 + and CD8 + T cell infiltrating tumor tissues of mice, bar: 100 μm. (I–K) The quantitative analysis of immune-related cytokines: IL-6, TNF-α and IFN-γ, ∗ p < 0.5, ∗∗ p < 0.01, ∗∗∗ p < 0.001 and ∗∗∗∗ p < 0.0001.

Article Snippet: The IFN-γ and TNF-α detection kit was purchased from Biodragon Biotechnology (Beijing), the IL-6 detection kit was obtained from BOSTER Biotechnology (Wuhan).

Techniques: In Vivo, Activation Assay, Flow Cytometry, Immunofluorescence

In vivo anti-tumor immune mechanisim analysis of BU-CDs CA-HA -loaded Janus membrane under NIR: (A–I) Flow cytometry and quantitative analysis of DCs, CD4 + /CD8 + T cells, NK cells and MDSCs, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; (J) Immunofluorescent staining of CD4 + /CD8 + T cells in tumor tissues to illustrate the enhanced tumor-specific identification and infiltration of T cells, bar: 50 μm; (K–L) Elisa test of proinflammatory cytokin (IL-6, TNF-α and IFN-γ) to assess the inflammatory activity, ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Journal: Materials Today Bio

Article Title: Immediate tumor killing and long-term anti-tumor immunoreaction induced by Bufalin-loaded phototherapeutic Janus membrane in CRC postoperative therapy

doi: 10.1016/j.mtbio.2026.102824

Figure Lengend Snippet: In vivo anti-tumor immune mechanisim analysis of BU-CDs CA-HA -loaded Janus membrane under NIR: (A–I) Flow cytometry and quantitative analysis of DCs, CD4 + /CD8 + T cells, NK cells and MDSCs, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; (J) Immunofluorescent staining of CD4 + /CD8 + T cells in tumor tissues to illustrate the enhanced tumor-specific identification and infiltration of T cells, bar: 50 μm; (K–L) Elisa test of proinflammatory cytokin (IL-6, TNF-α and IFN-γ) to assess the inflammatory activity, ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: The IFN-γ and TNF-α detection kit was purchased from Biodragon Biotechnology (Beijing), the IL-6 detection kit was obtained from BOSTER Biotechnology (Wuhan).

Techniques: In Vivo, Membrane, Flow Cytometry, Staining, Enzyme-linked Immunosorbent Assay, Activity Assay

HCP alleviates oxidative stress and systemic inflammatory response in HIRI. ( A ) Malondialdehyde (MDA) content in liver tissues at 24 h reperfusion ( n = 8). ( B ) Superoxide dismutase (SOD) activity in liver tissues at 24 h reperfusion ( n = 8). ( C , D ) Serum levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) at 24 h reperfusion ( n = 8). ( E ) Western blot analysis of inflammatory cytokine protein expression (IL-6, TNF-α, IL-1β) in liver tissues, with GAPDH as loading control (representative of three independent experiments). Data are mean ± SD. Statistical significance was determined by one-way ANOVA with Tukey’s post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. Vehicle group.

Journal: Biomedicines

Article Title: Houttuynia cordata Polysaccharide Alleviates Hepatic Ischemia-Reperfusion Injury by Regulating Macrophage Polarization via Inhibiting the TLR4/NF-κB Signaling Pathway

doi: 10.3390/biomedicines14020433

Figure Lengend Snippet: HCP alleviates oxidative stress and systemic inflammatory response in HIRI. ( A ) Malondialdehyde (MDA) content in liver tissues at 24 h reperfusion ( n = 8). ( B ) Superoxide dismutase (SOD) activity in liver tissues at 24 h reperfusion ( n = 8). ( C , D ) Serum levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) at 24 h reperfusion ( n = 8). ( E ) Western blot analysis of inflammatory cytokine protein expression (IL-6, TNF-α, IL-1β) in liver tissues, with GAPDH as loading control (representative of three independent experiments). Data are mean ± SD. Statistical significance was determined by one-way ANOVA with Tukey’s post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. Vehicle group.

Article Snippet: Mouse ALT, AST, MDA, SOD, TNF-α, and IL-6 ELISA detection kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).

Techniques: Activity Assay, Western Blot, Expressing, Control

HCP inhibits the TLR4/MyD88/NF-κB signaling pathway in HIRI, and its effects are reversed by TLR4 blockade. ( A ) Western blot analysis of TLR4, MyD88, total p65, and phospho-p65 (p-p65) protein expression in liver tissues at 24 h reperfusion. GAPDH served as loading control (representative blot of n = 3 independent samples/group). ( B ) Immunohistochemical staining for NF-κB p65 in liver sections (scale bar: 50 μm). Arrows indicate nuclear p65 staining. ( C – F ) Serum ALT, AST, TNF-α, and IL-6 levels at 24 h reperfusion in mice pretreated with or without the TLR4 inhibitor TAK-242 (3 mg/kg, i.p.) 1 h before ischemia ( n = 8). ( G ) Representative H&E-stained liver sections from corresponding groups (scale bar: 200 μm). ( H ) Western blot analysis of cytokine expression in liver tissues with or without TAK-242 intervention (representative of three experiments). Data are mean ± SD. Statistical analysis: one-way ANOVA with Tukey’s test. ** p < 0.01, **** p < 0.0001; ns, not significant.

Journal: Biomedicines

Article Title: Houttuynia cordata Polysaccharide Alleviates Hepatic Ischemia-Reperfusion Injury by Regulating Macrophage Polarization via Inhibiting the TLR4/NF-κB Signaling Pathway

doi: 10.3390/biomedicines14020433

Figure Lengend Snippet: HCP inhibits the TLR4/MyD88/NF-κB signaling pathway in HIRI, and its effects are reversed by TLR4 blockade. ( A ) Western blot analysis of TLR4, MyD88, total p65, and phospho-p65 (p-p65) protein expression in liver tissues at 24 h reperfusion. GAPDH served as loading control (representative blot of n = 3 independent samples/group). ( B ) Immunohistochemical staining for NF-κB p65 in liver sections (scale bar: 50 μm). Arrows indicate nuclear p65 staining. ( C – F ) Serum ALT, AST, TNF-α, and IL-6 levels at 24 h reperfusion in mice pretreated with or without the TLR4 inhibitor TAK-242 (3 mg/kg, i.p.) 1 h before ischemia ( n = 8). ( G ) Representative H&E-stained liver sections from corresponding groups (scale bar: 200 μm). ( H ) Western blot analysis of cytokine expression in liver tissues with or without TAK-242 intervention (representative of three experiments). Data are mean ± SD. Statistical analysis: one-way ANOVA with Tukey’s test. ** p < 0.01, **** p < 0.0001; ns, not significant.

Article Snippet: Mouse ALT, AST, MDA, SOD, TNF-α, and IL-6 ELISA detection kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).

Techniques: Western Blot, Expressing, Control, Immunohistochemical staining, Staining

HCP directly protects macrophages against hypoxia/reoxygenation (H/R) injury in a TLR4-dependent manner in vitro. ( A ) Cell viability of RAW264.7 macrophages assessed by CCK-8 assay after H/R (6 h hypoxia/12 h reoxygenation) with or without HCP co-treatment ( n = 6 independent wells per group). ( B , C ) ELISA measurement of TNF-α and IL-6 levels in culture supernatants after H/R ( n = 4 independent experiments). ( D ) Intracellular reactive oxygen species (ROS) levels measured by DCFH-DA fluorescence ( n = 4). ( E – G ) Corresponding TNF-α, IL-6, and ROS levels in cells pretreated with the TLR4 inhibitor TAK-242 (1 μM) 1 h prior to HCP and H/R ( n = 4). Data are mean ± SD. Statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. H/R (Vehicle) group; ns, not significant (one-way ANOVA with Tukey’s test).

Journal: Biomedicines

Article Title: Houttuynia cordata Polysaccharide Alleviates Hepatic Ischemia-Reperfusion Injury by Regulating Macrophage Polarization via Inhibiting the TLR4/NF-κB Signaling Pathway

doi: 10.3390/biomedicines14020433

Figure Lengend Snippet: HCP directly protects macrophages against hypoxia/reoxygenation (H/R) injury in a TLR4-dependent manner in vitro. ( A ) Cell viability of RAW264.7 macrophages assessed by CCK-8 assay after H/R (6 h hypoxia/12 h reoxygenation) with or without HCP co-treatment ( n = 6 independent wells per group). ( B , C ) ELISA measurement of TNF-α and IL-6 levels in culture supernatants after H/R ( n = 4 independent experiments). ( D ) Intracellular reactive oxygen species (ROS) levels measured by DCFH-DA fluorescence ( n = 4). ( E – G ) Corresponding TNF-α, IL-6, and ROS levels in cells pretreated with the TLR4 inhibitor TAK-242 (1 μM) 1 h prior to HCP and H/R ( n = 4). Data are mean ± SD. Statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. H/R (Vehicle) group; ns, not significant (one-way ANOVA with Tukey’s test).

Article Snippet: Mouse ALT, AST, MDA, SOD, TNF-α, and IL-6 ELISA detection kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).

Techniques: In Vitro, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Fluorescence

Schematic diagram illustrating the mechanism by which HCP alleviates HIRI by regulating macrophage polarization via the TLR4/NF-κB pathway. After three days of intraperitoneal injection pretreatment with HCP, a 70% hepatic ischemia for 1 h followed by 24 h reperfusion injury model was established. HCP inhibits the activation of TLR4 on the surface of hepatic Kupffer cells, and blocks the downstream MyD88-dependent NF-κB signaling pathway, thereby reducing NF-κB nuclear translocation and the release of pro-inflammatory cytokines (TNF-α, IL-6). Concurrently, HCP regulates macrophage phenotypes, suppressing M1 polarization and promoting M2 polarization, ultimately alleviating hepatic ischemia-reperfusion injury.

Journal: Biomedicines

Article Title: Houttuynia cordata Polysaccharide Alleviates Hepatic Ischemia-Reperfusion Injury by Regulating Macrophage Polarization via Inhibiting the TLR4/NF-κB Signaling Pathway

doi: 10.3390/biomedicines14020433

Figure Lengend Snippet: Schematic diagram illustrating the mechanism by which HCP alleviates HIRI by regulating macrophage polarization via the TLR4/NF-κB pathway. After three days of intraperitoneal injection pretreatment with HCP, a 70% hepatic ischemia for 1 h followed by 24 h reperfusion injury model was established. HCP inhibits the activation of TLR4 on the surface of hepatic Kupffer cells, and blocks the downstream MyD88-dependent NF-κB signaling pathway, thereby reducing NF-κB nuclear translocation and the release of pro-inflammatory cytokines (TNF-α, IL-6). Concurrently, HCP regulates macrophage phenotypes, suppressing M1 polarization and promoting M2 polarization, ultimately alleviating hepatic ischemia-reperfusion injury.

Article Snippet: Mouse ALT, AST, MDA, SOD, TNF-α, and IL-6 ELISA detection kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).

Techniques: Injection, Activation Assay, Translocation Assay