nmol l tumor necrosis factor  (R&D Systems)

Journal: Molecules

Article Title: Wound Healing Potential of the Salvianolic Acid H and Yunnaneic Acid B—The Rosmarinic Acid Derivatives: Anti-Inflammatory Action and Hemocompatibility In Vitro

doi: 10.3390/molecules31030452

Figure Lengend Snippet: Effects of the examined compounds: SA H, YA B, and RA on pro-inflammatory cytokine release in the concanavalin A (Con A)-stimulated PBMCs. The graphs refer to results from measurements of IL-1β, IL-6, and TNF-α secretion, respectively. DEX (dexamethasone)—a reference compound (an agonist of the glucocorticoid receptor—steroidal anti-inflammatory drug). The cytokine secretion from the Con A-stimulated cells, untreated with the examined phenolic acids, was assumed to be 100%. The figure represents mean values (±SD); * p < 0.05, ** p < 0.01; *** p < 0.001; n = 5.

Article Snippet: Recombinant human interferon-gamma (IFN-γ), recombinant human tumor necrosis factor-alpha (TNF-α), and lipoteichoic acid (LTA) from Gram-positive bacteria Staphylococcus aureus were purchased from InvivoGen (Toulouse, France).

Techniques:

Journal: Molecules

Article Title: Wound Healing Potential of the Salvianolic Acid H and Yunnaneic Acid B—The Rosmarinic Acid Derivatives: Anti-Inflammatory Action and Hemocompatibility In Vitro

doi: 10.3390/molecules31030452

Figure Lengend Snippet: Effects of SA H, YA B, and RA on the secretion of IL-6 and IL-8 by HaCaT cells (keratinocytes). Urolithin A (UroA)—a reference compound (25 µM). The figure represents mean values (±SD); ### p < 0.001—unstimulated HaCaT vs. HaCaT treated with TNF-α/interferon-gamma (IFN-γ) in the absence of the examined substances; the cytokine level (pg/mL) detected in samples derived from the TNF-α/IFN-γ -stimulated cells and treated with the examined acids vs. cells treated in the absence of the examined acids: * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Recombinant human interferon-gamma (IFN-γ), recombinant human tumor necrosis factor-alpha (TNF-α), and lipoteichoic acid (LTA) from Gram-positive bacteria Staphylococcus aureus were purchased from InvivoGen (Toulouse, France).

Techniques: Derivative Assay

Journal: Molecules

Article Title: Wound Healing Potential of the Salvianolic Acid H and Yunnaneic Acid B—The Rosmarinic Acid Derivatives: Anti-Inflammatory Action and Hemocompatibility In Vitro

doi: 10.3390/molecules31030452

Figure Lengend Snippet: Effects of SA H, YA B, and RA on the secretion of IL-6 and IL-8 by NHDF cells (fibroblasts). Urolithin A (UroA)—a reference compound (25 µM). The figure represents mean values (±SD); ### p < 0.001 refers to unstimulated NHDF vs. NHDF treated with TNF-α/IFN-γ in the absence of the examined substances; the cytokine level (pg/mL) detected in samples derived from the TNF-α/IFN-γ—stimulated cells and treated with the examined acids vs. cells treated in the absence of the examined acids: * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Recombinant human interferon-gamma (IFN-γ), recombinant human tumor necrosis factor-alpha (TNF-α), and lipoteichoic acid (LTA) from Gram-positive bacteria Staphylococcus aureus were purchased from InvivoGen (Toulouse, France).

Techniques: Derivative Assay

Journal: Frontiers in Immunology

Article Title: Dysregulated hyaluronan metabolism drives inflammation and angiogenesis in proliferative diabetic retinopathy

doi: 10.3389/fimmu.2026.1724199

Figure Lengend Snippet: Human retinal microvascular endothelial cells (HRMECs) were left untreated or treated with high-glucose (HG) (25 mM) for 24h. For HG treatment, cultures treated with mannitol (25 mM) were used as a control. Protein expression of hyaluronan synthase (HAS)2 (A) , hyaluronidase (Hyal)-1 (B) , Hyal-2 (C) and CD44 (D) in cell lysate was determined by Western blot analysis. (E) HRMECs were left untreated or treated with tumor necrosis factor–α (TNF-α) (5ng/ml) or hydrogen peroxide (H 2 O 2 ) (10mM) for 24h. Levels of Hyal-1 were quantified in the culture media by ELISA. (F) HRMECs and human retinal Müller glial cells were compared after treatment with mannitol (25 mM) or HG (25 mM) for 24h. Levels of hyaluronan (HA) were quantified in the culture media by ELISA. Results are expressed as mean ± standard deviation from three different experiments each performed in triplicate (*p < 0.05; independent t-test).

Article Snippet: The following stimuli were used to mimic diabetic conditions: treatment of Müller cells or HRMECs with 5 ng/ml recombinant human tumor necrosis factor-alpha (TNF-α) (Cat No 210-TA, R&D Systems), 300 μM of the hypoxia mimetic agent cobalt chloride (CoCl 2 ) (Cat No A1425-L, Avonchem Limited, UK), or 10 mM H 2 O 2 , or 25mM glucose (Cat No GL0125100, Scharlau S.L, Gato Prez, Spain) for 16 h. To compensate for osmotic effects when using 25mM glucose as high-glucose (HG) treatment, 25 mM mannitol (Cat No MA01490500, Scharlau S.L, Gato Prez, Spain) was used as a control.

Techniques: Control, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Frontiers in Immunology

Article Title: Dysregulated hyaluronan metabolism drives inflammation and angiogenesis in proliferative diabetic retinopathy

doi: 10.3389/fimmu.2026.1724199

Figure Lengend Snippet: Human retinal Müller glial cells were left untreated or treated with ultra-low molecular weight hyaluronan (ULMW-HA) (50 µg/mL) for 24 (h) (A) Protein expression of phospho-ERK1/2 and phospho-NFκB in cell lysates was determined by Western blot analysis. Levels of high mobility group box-1 (HMGB1) were quantified in the culture media by ELISA. Results are expressed as mean ± standard deviation from three different experiments each performed in triplicate (*p < 0.05; independent t-test). (B) Human retinal Müller glial cells were left untreated or treated with ULMW-HA, ULMW-HA plus BAY11-7085 (5 µM) or (C) ULMW-HA plus U-0126 (5 µM). Levels of vascular endothelial growth factor (VEGF), angiopoietin and monocyte chemotactic protein-1 (MCP-1/CCL2) were quantified in the culture media by ELISA. Results are expressed as mean ± standard deviation or standard error of mean from three different experiments each performed in triplicate. One-way ANOVA and independent t-test were used for comparisons between three groups and two groups, respectively. *p < 0.05 compared with values obtained from untreated cells; #p < 0.05 compared with ULMW-HA plus BAY11–7085 or U-0126 treated cells. (D, E) Human retinal Müller glial cells were left untreated or treated with high glucose (HG) (25 mM), cobalt chloride (CoCl 2 ) (300 µM) or tumor necrosis factor-α (TNF-α) (5 ng/mL) with or without apigenin (10 µg/mL) for 24 (h) For HG treatment, cultures containing 25 mM mannitol were used as a control. Levels of monocyte chemotactic protein-1 (MCP-1/CCL2) (D) and vascular endothelial growth factor (VEGF) (E) were quantified in the culture media by ELISA. The results are expressed as mean ± standard deviation from three different experiments each performed in triplicate. One-way ANOVA and independent t-test were used for comparisons between three and two groups, respectively. *p < 0.05 compared with values obtained from control cells. #p < 0.05 compared with values obtained from stimulated cells.

Article Snippet: The following stimuli were used to mimic diabetic conditions: treatment of Müller cells or HRMECs with 5 ng/ml recombinant human tumor necrosis factor-alpha (TNF-α) (Cat No 210-TA, R&D Systems), 300 μM of the hypoxia mimetic agent cobalt chloride (CoCl 2 ) (Cat No A1425-L, Avonchem Limited, UK), or 10 mM H 2 O 2 , or 25mM glucose (Cat No GL0125100, Scharlau S.L, Gato Prez, Spain) for 16 h. To compensate for osmotic effects when using 25mM glucose as high-glucose (HG) treatment, 25 mM mannitol (Cat No MA01490500, Scharlau S.L, Gato Prez, Spain) was used as a control.

Techniques: Molecular Weight, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Standard Deviation, Control

Journal: Frontiers in Immunology

Article Title: Dysregulated hyaluronan metabolism drives inflammation and angiogenesis in proliferative diabetic retinopathy

doi: 10.3389/fimmu.2026.1724199

Figure Lengend Snippet: Human retinal microvascular endothelial cells (HRMECs) were left untreated or stimulated with tumor necrosis factor-α (TNF-α) (5 ng/mL) for 24 h with or without apigenin (10 µg/mL). Protein expression of intercellular adhesion molecule-1 (ICAM-1) (A) and vascular cell adhesion molecule-1 (VCAM-1) (B) was determined by Western blot analysis. Adhesion of fluorescently labeled THP-1 monocytic cells to HRMECs monolayer was quantified (C) . Results are expressed as mean ± standard deviation or standard error of mean from three different experiments each performed in triplicate. One-way ANOVA and independent t-test were used for comparisons between three groups and two groups, respectively. *p < 0.05 compared with values obtained from untreated cells. #p < 0.05 compared with values obtained from cells treated with TNF-α (RFU = relative fluorescence unit).

Article Snippet: The following stimuli were used to mimic diabetic conditions: treatment of Müller cells or HRMECs with 5 ng/ml recombinant human tumor necrosis factor-alpha (TNF-α) (Cat No 210-TA, R&D Systems), 300 μM of the hypoxia mimetic agent cobalt chloride (CoCl 2 ) (Cat No A1425-L, Avonchem Limited, UK), or 10 mM H 2 O 2 , or 25mM glucose (Cat No GL0125100, Scharlau S.L, Gato Prez, Spain) for 16 h. To compensate for osmotic effects when using 25mM glucose as high-glucose (HG) treatment, 25 mM mannitol (Cat No MA01490500, Scharlau S.L, Gato Prez, Spain) was used as a control.

Techniques: Expressing, Western Blot, Labeling, Standard Deviation, Fluorescence

Journal: Frontiers in Immunology

Article Title: Dysregulated hyaluronan metabolism drives inflammation and angiogenesis in proliferative diabetic retinopathy

doi: 10.3389/fimmu.2026.1724199

Figure Lengend Snippet: Human retinal microvascular endothelial cells (HRMECs) were left untreated or treated with high glucose (HG) (25 mM) (A) , cobalt chloride (CoCl 2 ) (300 µM) (B) or tumor necrosis factor-α (TNF-α) (5 ng/mL) (C) with or without apigenin (10 µg/mL). For HG treatment, cultures treated with mannitol (25 mM) were used as a control. Levels of soluble syndecan-1 were quantified in the culture media by ELISA. Results are expressed as mean ± standard deviation from three different experiments each performed in triplicate. One-way ANOVA and independent t-test were used for comparisons between three and two groups, respectively. *p < 0.05 compared with values obtained from control cells. #p < 0.05 compared with values obtained from cells treated with HG, CoCl 2 or TNF-α. HRMECs were left untreated or were stimulated with ultra-low molecular weight – hyaluronan (ULMW-HA) (50 µg/mL) for 24 (h). Protein expression of phospho-ERK1/2 in the cell lysates was determined by Western blot analysis (D) . Results are expressed as mean ± standard deviation from three different experiments each performed in triplicate (*p < 0.05; independent t-test). A scratch was performed in confluent monolayers of overnight starved HRMECs with a micropipette tip subsequently, the cultures were left untreated or treated either with VEGF (10 ng/mL) or with ULMW-HA (100 µg/mL) for 16 (h) Cells were visualized using an inverted microscope. Two independent experiments were performed. Each experiment was done in duplicate, and 2–3 independent field images were taken for the migration analysis which was done by using Image J software. In the Figure, one representative image is illustrated, and the bar graphs show the analysis of all the images from each group represented as fold-change in migration versus control (E) . Results are expressed as mean ± standard deviation. One-way ANOVA and independent t-test were used for comparisons between three and two groups, respectively. *p < 0.05 compared with values obtained from control cells.

Article Snippet: The following stimuli were used to mimic diabetic conditions: treatment of Müller cells or HRMECs with 5 ng/ml recombinant human tumor necrosis factor-alpha (TNF-α) (Cat No 210-TA, R&D Systems), 300 μM of the hypoxia mimetic agent cobalt chloride (CoCl 2 ) (Cat No A1425-L, Avonchem Limited, UK), or 10 mM H 2 O 2 , or 25mM glucose (Cat No GL0125100, Scharlau S.L, Gato Prez, Spain) for 16 h. To compensate for osmotic effects when using 25mM glucose as high-glucose (HG) treatment, 25 mM mannitol (Cat No MA01490500, Scharlau S.L, Gato Prez, Spain) was used as a control.

Techniques: Control, Enzyme-linked Immunosorbent Assay, Standard Deviation, Molecular Weight, Expressing, Western Blot, Inverted Microscopy, Migration, Software