Journal: Pharmaceuticals

Article Title: CB2 Receptor Activation Attenuates IL-1β-Induced Inflammatory Transcriptomic Networks in Human Gingival Fibroblasts

doi: 10.3390/ph19091406

Figure Lengend Snippet: HU-308 attenuates IL-1β-induced interleukin expression in human gingival fibroblasts. Human gingival fibroblasts (HGFs) were divided into three experimental groups: untreated Control (black bars), IL-1β (10 ng/mL)-stimulated cells (red bars), and cells treated with IL-1β followed by the selective CB2 receptor agonist HU-308 (10 μM; blue bars). Twenty-four hours after IL-1β stimulation, transcript levels of proinflammatory and anti-inflammatory interleukins were quantified. IL-1β stimulation significantly increased the expression of the pro-inflammatory cytokines IL1B, IL6, IL23A, IL33, IL11, IL1α, and IL24, and treatment with HU-308 attenuated the IL-1β-induced expression of these cytokines, indicating suppression of the inflammatory response ( A – F ). In contrast, IL10 exhibited reduced expression following IL-1β stimulation, and HU-308 treatment partially restored its expression, supporting a shift toward a less inflammatory phenotype following CB2 receptor activation ( G ). Data are presented as mean ± SEM from three independent biological replicates ( n = 3 per group). Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple-comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 was considered statistically significant.

Article Snippet: Recombinant human IL-1β was obtained from (InvivoGen, San Diego, CA, USA), reconstituted in water according to the manufacturer’s instructions, and used at a final concentration of 10 ng/mL.

Techniques: Expressing, Control, Activation Assay, Comparison

Journal: Pharmaceuticals

Article Title: CB2 Receptor Activation Attenuates IL-1β-Induced Inflammatory Transcriptomic Networks in Human Gingival Fibroblasts

doi: 10.3390/ph19091406

Figure Lengend Snippet: HU-308 modulates IL-1β-induced chemokine expression in human gingival fibroblasts. Human gingival fibroblasts (HGFs) were divided into three experimental groups: untreated Control (black bars), IL-1β (10 ng/mL)-stimulated cells (red bars), and cells treated with IL-1β followed by the selective CB2 receptor agonist HU-308 (10 μM; blue bars). Twenty-four hours after IL-1β stimulation, transcript levels of inflammatory chemokines were quantified. IL-1β stimulation significantly increased the expression of CXCL5, CXCL8, CXCL16, CCL20, and CCL28, consistent with activation of chemokine-mediated inflammatory signaling ( A – E ). Treatment with HU-308 attenuated the IL-1β-induced expression of these chemokines, indicating suppression of inflammatory chemokine responses and reduced potential for immune-cell recruitment ( A – E ). Data are presented as mean ± SEM from three independent biological replicates ( n = 3 per group). Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple-comparison test. * p < 0.05, *** p < 0.001, and **** p < 0.0001 was considered statistically significant.

Article Snippet: Recombinant human IL-1β was obtained from (InvivoGen, San Diego, CA, USA), reconstituted in water according to the manufacturer’s instructions, and used at a final concentration of 10 ng/mL.

Techniques: Expressing, Control, Activation Assay, Immune Cell, Comparison

Journal: Pharmaceuticals

Article Title: CB2 Receptor Activation Attenuates IL-1β-Induced Inflammatory Transcriptomic Networks in Human Gingival Fibroblasts

doi: 10.3390/ph19091406

Figure Lengend Snippet: HU-308 attenuates IL-1β-induced matrix metalloproteinase expression in human gingival fibroblasts. Human gingival fibroblasts (HGFs) were divided into three experimental groups: untreated Control (black bars), IL-1β (10 ng/mL)-stimulated cells (red bars), and cells treated with IL-1β followed by the selective CB2 receptor agonist HU-308 (10 μM; blue bars). Twenty-four hours after IL-1β stimulation, transcript levels of MMP1, MMP3, and MMP10 were quantified. IL-1β stimulation significantly increased the expression of all three matrix metalloproteinases, consistent with enhanced extracellular matrix degradation and tissue remodeling associated with periodontal inflammation ( A – C ). Treatment with HU-308 significantly attenuated the IL-1β-induced expression of MMP1, MMP3, and MMP10, suggesting that CB2 receptor modulation suppresses matrix-degrading pathways and may contribute to the preservation of periodontal tissue integrity ( A – C ). Data are presented as mean ± SEM from three independent biological replicates ( n = 3 per group). Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple-comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 was considered statistically significant.

Article Snippet: Recombinant human IL-1β was obtained from (InvivoGen, San Diego, CA, USA), reconstituted in water according to the manufacturer’s instructions, and used at a final concentration of 10 ng/mL.

Techniques: Expressing, Control, Tissue, Preserving, Comparison

Journal: Pharmaceuticals

Article Title: CB2 Receptor Activation Attenuates IL-1β-Induced Inflammatory Transcriptomic Networks in Human Gingival Fibroblasts

doi: 10.3390/ph19091406

Figure Lengend Snippet: HU-308 partially restores IL-1β-induced alterations in collagen gene expression in human gingival fibroblasts. Human gingival fibroblasts (HGFs) were divided into three experimental groups: untreated Control (black bars), IL-1β (10 ng/mL)-stimulated cells (red bars), and cells treated with IL-1β followed by the selective CB2 receptor agonist HU-308 (10 μM; blue bars). Twenty-four hours after IL-1β stimulation, transcript levels of COL1A1, COL3A1, COL11A1, and COL12A1 were quantified. IL-1β stimulation significantly reduced the expression of collagen-associated genes, consistent with impaired extracellular matrix synthesis and tissue remodeling during periodontal inflammation ( A – D ). Treatment with HU-308 partially restored the expression of these collagen transcripts toward control levels, suggesting that CB2 receptor activation may preserve extracellular matrix homeostasis and limit IL-1β-induced connective tissue damage ( A – D ). Data are presented as mean ± SEM from three independent biological replicates ( n = 3 per group). Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple-comparison test. * p < 0.05, ** p < 0.01 and *** p < 0.001 was considered statistically significant.

Article Snippet: Recombinant human IL-1β was obtained from (InvivoGen, San Diego, CA, USA), reconstituted in water according to the manufacturer’s instructions, and used at a final concentration of 10 ng/mL.

Techniques: Gene Expression, Control, Expressing, Tissue, Activation Assay, Comparison

Journal: Pharmaceuticals

Article Title: CB2 Receptor Activation Attenuates IL-1β-Induced Inflammatory Transcriptomic Networks in Human Gingival Fibroblasts

doi: 10.3390/ph19091406

Figure Lengend Snippet: HU-308 modulates IL-1β-induced nitric oxide and oxidative stress-related gene expression in human gingival fibroblasts. Human gingival fibroblasts (HGFs) were divided into three experimental groups: untreated Control (black bars), IL-1β (10 ng/mL)-stimulated cells (red bars), and cells treated with IL-1β followed by the selective CB2 receptor agonist HU-308 (10 μM; blue bars). Twenty-four hours after IL-1β stimulation, transcript levels of NOS3, PRKG1 and PDE5A were quantified. IL-1β stimulation significantly altered the expression of genes involved in nitric oxide signaling, reflecting activation of redox-associated inflammatory pathways ( A – C ). Treatment with HU-308 modulated the IL-1β-induced expression of these genes, partially restoring their expression toward control levels and suggesting that CB2 receptor activation attenuates redox imbalance and nitric oxide signaling dysregulation associated with inflammatory activation ( A – C ). Data are presented as mean ± SEM from three independent biological replicates ( n = 3 per group). Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple-comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 was considered statistically significant.

Article Snippet: Recombinant human IL-1β was obtained from (InvivoGen, San Diego, CA, USA), reconstituted in water according to the manufacturer’s instructions, and used at a final concentration of 10 ng/mL.

Techniques: Gene Expression, Control, Expressing, Activation Assay, Comparison

Journal: Pharmaceuticals

Article Title: CB2 Receptor Activation Attenuates IL-1β-Induced Inflammatory Transcriptomic Networks in Human Gingival Fibroblasts

doi: 10.3390/ph19091406

Figure Lengend Snippet: HU-308 modulates IL-1β-associated changes in solute carrier transporter transcript expression in human gingival fibroblasts. Human gingival fibroblasts (HGFs) were divided into three experimental groups: untreated Control (black bars), IL-1β (10 ng/mL)-stimulated cells (red bars), and cells treated with IL-1β followed by the selective CB2 receptor agonist HU-308 (10 μM; blue bars). Twenty-four hours after IL-1β stimulation, transcript levels of SLC2A1, SLC2A3, SLC2A5, SLC5A13, and SLC2A14 were quantified. IL-1β stimulation significantly altered the expression of SLC2A1, SLC2A3, SLC2A5, SLC2A13, and SLC2A14, and HU-308 treatment significantly modified several of these IL-1β-associated transcriptional responses ( A – E ). These findings demonstrate changes in transporter-associated transcript abundance but do not establish corresponding alterations in substrate uptake, glucose utilization, glycolytic activity, or cellular metabolic flux. Data are presented as mean ± SEM from three independent biological replicates ( n = 3 per group). Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple-comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 was considered statistically significant.

Article Snippet: Recombinant human IL-1β was obtained from (InvivoGen, San Diego, CA, USA), reconstituted in water according to the manufacturer’s instructions, and used at a final concentration of 10 ng/mL.

Techniques: Expressing, Control, Modification, Activity Assay, Comparison

Journal: Pharmaceuticals

Article Title: CB2 Receptor Activation Attenuates IL-1β-Induced Inflammatory Transcriptomic Networks in Human Gingival Fibroblasts

doi: 10.3390/ph19091406

Figure Lengend Snippet: GPCR-related transcript modulation by IL-1β and HU-308 treatment in human gingival fibroblasts. Human gingival fibroblasts (HGFs) were divided into three experimental groups: untreated Control (black bars), IL-1β (10 ng/mL)-stimulated cells (red bars), and cells treated with IL-1β followed by the selective CB2 receptor agonist HU-308 (10 μM; blue bars). Twenty-four hours after IL-1β stimulation, transcript levels of GPR160, GPR180, GPR157, GPR61, GPR173, GPR155, GPR107, GPR75, GPR78, and GPR4 were quantified. IL-1β stimulation significantly altered the expression of multiple GPCR-related genes, including increased expression of GPR160, GPR180, GPR157, GPR61, GPR173, GPR155, GPR107, and GPR75, while reducing the expression of GPR78 and GPR4, indicating inflammatory remodeling of GPCR-associated signaling pathways in human gingival fibroblasts ( A – J ). Treatment with HU-308 modified several IL-1β-associated changes in GPCR-related transcript expression ( A – J ). These changes represent transcriptional associations and should not be interpreted as evidence of altered receptor activity or direct functional cross-talk with CB2. Data are presented as mean ± SEM from three independent biological replicates ( n = 3 per group). Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple-comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 was considered statistically significant.

Article Snippet: Recombinant human IL-1β was obtained from (InvivoGen, San Diego, CA, USA), reconstituted in water according to the manufacturer’s instructions, and used at a final concentration of 10 ng/mL.

Techniques: Control, Expressing, Protein-Protein interactions, Modification, Activity Assay, Functional Assay, Comparison

Journal: Pharmaceuticals

Article Title: CB2 Receptor Activation Attenuates IL-1β-Induced Inflammatory Transcriptomic Networks in Human Gingival Fibroblasts

doi: 10.3390/ph19091406

Figure Lengend Snippet: Schematic summary of the coordinated anti-inflammatory effects of CB2 receptor activation in IL-1β-stimulated human gingival fibroblasts. IL-1β stimulation establishes a pro-inflammatory transcriptional landscape characterized by coordinated upregulation of inflammatory cytokines, chemokines, matrix metalloproteinases (MMPs), glucose transporter (SLC2/GLUT) genes, and multiple G protein-coupled receptor (GPCR)-related transcripts, together with suppression of collagen-associated genes and nitric oxide (NO)/cGMP signaling components. Collectively, these changes promote inflammatory amplification, extracellular matrix degradation, metabolic reprogramming, impaired tissue repair, and disruption of periodontal homeostasis ( left panel ). Pharmacological modulation of the cannabinoid receptor type 2 (CB2) with the selective agonist HU-308 shifts this inflammatory landscape toward a more homeostatic phenotype (center panel). HU-308 broadly attenuates IL-1β-induced inflammatory responses by reducing the expression of pro-inflammatory cytokines, chemokines, MMPs, glucose transporter-related genes, and several GPR-associated transcripts, while restoring collagen gene expression, NO/cGMP signaling, and selected receptor-associated pathways ( right panel ). Upward and downward arrows beneath each biological domain summarize the direction of transcript changes for individual genes following IL-1β stimulation (red arrows) and subsequent HU-308 treatment (blue arrows). Overall, the integrated pathway analysis demonstrates that CB2 modulation functions as an upstream regulator of multiple interconnected biological networks, including inflammatory signaling, extracellular matrix remodeling, redox homeostasis, metabolic adaptation, and GPR-associated signaling, rather than acting on a single molecular pathway, thereby supporting a systems-level mechanism through which CB2 modulation may preserve periodontal tissue homeostasis during inflammation. This figure represents a conceptual summary of selected transcriptional patterns and does not constitute a statistically derived gene-network, pathway-enrichment, or genome-wide differential-expression analysis.

Article Snippet: Recombinant human IL-1β was obtained from (InvivoGen, San Diego, CA, USA), reconstituted in water according to the manufacturer’s instructions, and used at a final concentration of 10 ng/mL.

Techniques: Activation Assay, Amplification, Tissue, Disruption, Expressing, Gene Expression, Analysis, Derivative Assay, Genome Wide, Quantitative Proteomics

Journal: Frontiers in Physiology

Article Title: Interleukin-13 R130Q aggravates airway smooth-muscle phenotypic switching and promotes airway remodeling in severe asthma

doi: 10.3389/fphys.2026.1906184

Figure Lengend Snippet: Effects of IL-13 130R and IL-13 130Q on pulmonary inflammatory responses. (A) Total cell counts in bronchoalveolar lavage fluid (BALF; n = 8 per group); differential leukocyte counting was not performed. (B–D) Relative mRNA expression levels of IL-4 (B) , IL-5 (C) , and IL-13 (D) in lung tissue (n = 8 per group). Data are presented as the mean ± SEM. *P < 0.05, **P < 0.01. IL, interleukin; OVA, ovalbumin; SEM, standard error of the mean.

Article Snippet: According to the instructions of the manufacturer for the IL-4 and IL-5 ELISA kits (RayBiotech, Peachtree Corners, GA, USA), 100 μL of each sample was added to pre-coated wells and incubated at room temperature for 2.5 h. The wells were then washed three times for 5 min each, and blank wells served as negative controls.

Techniques: Expressing

Journal: Frontiers in Physiology

Article Title: Interleukin-13 R130Q aggravates airway smooth-muscle phenotypic switching and promotes airway remodeling in severe asthma

doi: 10.3389/fphys.2026.1906184

Figure Lengend Snippet: Effects of IL-13 130R and IL-13 130Q on human bronchial smooth muscle cell (hBSMC) phenotypes (n = 4 per group). (A) CCK-8-derived proliferative/metabolic activity. (B) Representative Fluo-3/AM flow-cytometry histograms and median fluorescence intensity quantification of the acetylcholine-induced intracellular calcium response. (C, D) IL-5 (C) and IL-4 (D) concentrations in culture supernatants. Data are presented as the mean ± SEM. *P < 0.05, **P < 0.01. IL, interleukin; SEM, standard error of the mean.

Article Snippet: According to the instructions of the manufacturer for the IL-4 and IL-5 ELISA kits (RayBiotech, Peachtree Corners, GA, USA), 100 μL of each sample was added to pre-coated wells and incubated at room temperature for 2.5 h. The wells were then washed three times for 5 min each, and blank wells served as negative controls.

Techniques: CCK-8 Assay, Derivative Assay, Activity Assay, Flow Cytometry, Fluorescence

Journal: bioRxiv

Article Title: Complex Modulation of IL-6 Signaling by Apelin and Elabela in HTR-8/SVneo Cells Under Cobalt Chloride-Induced Chemical Hypoxia

doi: 10.64898/2026.08.20.746041

Figure Lengend Snippet: The effect of CoCl 2 on the levels of elabela (ELA), apelin, and apelin receptor APJ. The data were obtained using ELA or apelin ELISA assays in HTR-8/SVneo cell homogenates (Panels A and C) or conditioned media (Panels B and D), western blotting (Panel E), or real-time RT-PCR (Panel F) for APJ. Data are mean ± SEM, n=5-6.

Article Snippet: Individual ELISA kits were used for IL-6 (RayBiotech Inc., Norcross, GA, USA; cat. No. ELH-IL6), sIL-6R (RayBiotech Inc., Norcross, GA, USA; cat. No. ELH-IL6sR), and sgp130 (RayBiotech Inc., Norcross, GA, USA; cat. No. ELH-sgp130) protein quantification in the conditioned media 24 hours after treatment.

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

Journal: bioRxiv

Article Title: Complex Modulation of IL-6 Signaling by Apelin and Elabela in HTR-8/SVneo Cells Under Cobalt Chloride-Induced Chemical Hypoxia

doi: 10.64898/2026.08.20.746041

Figure Lengend Snippet: Elabela (ELA) or apelin treatment decreased IL-6 mRNA in CoCl 2 -treated HTR-8/SVneo cells. Data are mean ± SEM, n=3-6.

Article Snippet: Individual ELISA kits were used for IL-6 (RayBiotech Inc., Norcross, GA, USA; cat. No. ELH-IL6), sIL-6R (RayBiotech Inc., Norcross, GA, USA; cat. No. ELH-IL6sR), and sgp130 (RayBiotech Inc., Norcross, GA, USA; cat. No. ELH-sgp130) protein quantification in the conditioned media 24 hours after treatment.

Techniques:

Journal: bioRxiv

Article Title: Complex Modulation of IL-6 Signaling by Apelin and Elabela in HTR-8/SVneo Cells Under Cobalt Chloride-Induced Chemical Hypoxia

doi: 10.64898/2026.08.20.746041

Figure Lengend Snippet: Schematic of proposed interactions between IL-6 signaling and apelin and elabela in HTR-8/SVneo trophoblastic cells under CoCl₂-mimicked hypoxia. We hypothesize that apelin/elabela/APJ axis regulates IL-6 signaling via its actions on various components of the IL-6 classical and trans-signaling pathway.

Article Snippet: Individual ELISA kits were used for IL-6 (RayBiotech Inc., Norcross, GA, USA; cat. No. ELH-IL6), sIL-6R (RayBiotech Inc., Norcross, GA, USA; cat. No. ELH-IL6sR), and sgp130 (RayBiotech Inc., Norcross, GA, USA; cat. No. ELH-sgp130) protein quantification in the conditioned media 24 hours after treatment.

Techniques: