Journal: Scientific Reports

Article Title: Osthol ameliorates obesity-associated lipid metabolic disorders by inhibiting ADRA1D-dependent Th17 cell differentiation

doi: 10.1038/s41598-025-20719-x

Figure Lengend Snippet: OST Attenuates Systemic Inflammation and Downregulates Th17-Related Factor Expression. ( A ) KEGG pathway enrichment analysis of downstream targets of OST. This figure is based on data from KEGG (Kyoto Encyclopedia of Genes and Genomes) at https://www.kegg.jp/ ; © Kanehisa Laboratories. ( B ) Flow cytometry was used to evaluate the proportion of CD4⁺IL-17A⁺ cells in the spleen. ( C ) Immunohistochemical staining of liver tissues was performed to assess IL-17A expression. ( D ) Immunofluorescence staining was conducted to detect CD4⁺RORγt⁺ double-positive cells in liver sections. ( E ) Serum levels of IL-6, IL-1β, TNF-α, and IL-17A were measured using ELISA. n = 6. **** P < 0.0001. One-way ANOVA followed by Tukey’s post hoc test was used for multiple group comparisons.

Article Snippet: Cells were then fixed and permeabilized, followed by intracellular staining with a phycoerythrin (PE)-conjugated anti-IL-17A antibody (F21IL1702, Lianke Biotech, Hangzhou, China).

Techniques: Expressing, Flow Cytometry, Immunohistochemical staining, Staining, Immunofluorescence, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Osthol ameliorates obesity-associated lipid metabolic disorders by inhibiting ADRA1D-dependent Th17 cell differentiation

doi: 10.1038/s41598-025-20719-x

Figure Lengend Snippet: OST Inhibits Th17 Cell Differentiation in Vitro. ( A ) Flow cytometry was used to assess the proportion of CD4⁺IL-17A⁺ cells in a Th17-polarized cell model treated with different doses of OST. ( B ) RT-qPCR was performed to evaluate mRNA expression levels of RORγt and IL-17A. ( C ) Western blot was performed to evaluate protein expression levels of RORγt, IL-17A, IL-17RA, TRAF6, and Act1. ( D ) ELISA was used to quantify the secretion of IL-6, IL-1β, TNF-α, IL-10, and TGF-β in the culture supernatants. n = 3. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. One-way ANOVA followed by Tukey’s post hoc test was used for multiple group comparisons.

Article Snippet: Cells were then fixed and permeabilized, followed by intracellular staining with a phycoerythrin (PE)-conjugated anti-IL-17A antibody (F21IL1702, Lianke Biotech, Hangzhou, China).

Techniques: Cell Differentiation, In Vitro, Flow Cytometry, Quantitative RT-PCR, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Osthol ameliorates obesity-associated lipid metabolic disorders by inhibiting ADRA1D-dependent Th17 cell differentiation

doi: 10.1038/s41598-025-20719-x

Figure Lengend Snippet: OST Suppresses Th17 Differentiation via an ADRA1D-Dependent Mechanism. ( A ) Volcano plot of differentially expressed genes (DEGs) in the GSE110729 obesity dataset. ( B ) Venn diagram showing the intersection between OST’s potential targets and DEGs from the GSE110729 dataset. ( C ) RT-qPCR was performed to assess the expression of ADRA1D, IL-17A, and RORγt in Th17-polarized CD4⁺ T cells with or without ADRA1D overexpression. ( D ) Western blot was conducted to evaluate the protein expression of ADRA1D, IL-17A, RORγt, IL-17RA, TRAF6, and Act1. ( E ) ELISA was used to measure the levels of IL-6, IL-1β, TNF-α, IL-10, and TGF-β in the culture supernatants. n = 3. *** P < 0.001, **** P < 0.0001. One-way ANOVA followed by Tukey’s post hoc test was used for multiple group comparisons.

Article Snippet: Cells were then fixed and permeabilized, followed by intracellular staining with a phycoerythrin (PE)-conjugated anti-IL-17A antibody (F21IL1702, Lianke Biotech, Hangzhou, China).

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

Journal: Scientific Reports

Article Title: Osthol ameliorates obesity-associated lipid metabolic disorders by inhibiting ADRA1D-dependent Th17 cell differentiation

doi: 10.1038/s41598-025-20719-x

Figure Lengend Snippet: ADRA1D Overexpression Attenuates the Metabolic and Immunomodulatory Effects of OST in Vivo. ( A ) Body weight changes were monitored in HFD-fed mice treated with OST. (B) Weights of adipose tissue and liver were measured to assess the effect on organ hypertrophy. ( C ) Serum levels of TG, TC, FFA, ALT, and AST were evaluated by biochemical assays. ( D ) Flow cytometry was performed to assess the proportion of CD4⁺IL-17A⁺ cells in the spleen. ( E ) Immunofluorescence staining of liver tissue was used to detect CD4⁺RORγt⁺ double-positive cell infiltration. ( F ) Immunohistochemistry was performed to evaluate hepatic IL-17A protein expression. ( G ) Western blotting was conducted to assess ADRA1D protein expression in liver tissue. ( H ) Western blot was used to detect the phosphorylation levels of ERK1/2 and PI3K in liver tissue. ( I ) H&E staining of adipose tissue was used to assess adipocyte morphology. (J) Oil red O staining of liver sections was performed to evaluate hepatic lipid deposition. n = 6. ** P < 0.01, **** P < 0.0001. For three or more groups, one-way or two-way analysis of variance (ANOVA) was applied, followed by Tukey’s post hoc test. P < 0.05 was considered statistically significant.

Article Snippet: Cells were then fixed and permeabilized, followed by intracellular staining with a phycoerythrin (PE)-conjugated anti-IL-17A antibody (F21IL1702, Lianke Biotech, Hangzhou, China).

Techniques: Over Expression, In Vivo, Flow Cytometry, Immunofluorescence, Staining, Immunohistochemistry, Expressing, Western Blot, Phospho-proteomics

Journal: Vaccines

Article Title: Mixed Th1/Th2/Th17 Responses Induced by Plant Oil Adjuvant-Based B. bronchiseptica Vaccine in Mice, with Mechanisms Unraveled by RNA-Seq, 16S rRNA and Metabolomics

doi: 10.3390/vaccines12101182

Figure Lengend Snippet: Cytokine production in serum (Mice, n = 5/group). Serum samples were collected 14 days after the boost and detected by a cytokine array. ( A ) Heat map of hierarchical clustering of cytokines in Bb vs. alum. ( B ) Heat map of hierarchical clustering of cytokines in Bb vs. E515. ( C ) Heat map of hierarchical clustering of cytokines in alum vs. E515. ( D ) KEGG pathway enrichment of cytokines in Bb vs. alum. ( E ) KEGG pathway enrichment of cytokines in Bb vs. E515. ( F ) KEGG pathway enrichment of cytokines in alum vs. E515. Red color on the heatmap indicates cytokines are upregulated and blue indicates cytokines are downregulated. Dot size on the KEGG pathway represents the number of cytokines; the color represents the p value.

Article Snippet: After centrifugation (1500 rpm, 5 min), the supernatant was collected for the measurement of cytokines (IFN-γ, IL-6, TGF-β1 and IL-17) by commercial ELISA kits (Multi-Sciences Biotech, Hangzhou, China) in accordance with the manufacturer’s instructions.

Techniques: