Journal: Molecules

Article Title: Lignans from Machilus thunbergii as Thymic Stromal Lymphopoietin Inhibitors

doi: 10.3390/molecules26164804

Figure Lengend Snippet: The inhibitory effects of isolated compounds 1 – 3 on thymic stromal lymphopoietin (TSLP)/TSLP receptor (TSLPR) interaction by ELISA.

Article Snippet: For the pSTAT5 assay measuring TSLP inhibitory activity, recombinant human TSLP was purchased from R&D Systems (Minneapolis, MN, USA) and BD Cytofix/Cytoperm and Alexa Fluor 647 mouse anti-STAT5 (pY694) were purchased from BD Biosciences (San Diego, CA, USA).

Techniques: Isolation, Enzyme-linked Immunosorbent Assay, Inhibition, Control

Journal: Molecules

Article Title: Lignans from Machilus thunbergii as Thymic Stromal Lymphopoietin Inhibitors

doi: 10.3390/molecules26164804

Figure Lengend Snippet: Molecular docking results of isolated compounds 1 – 3 on TSLP/TSLPR interaction.

Article Snippet: For the pSTAT5 assay measuring TSLP inhibitory activity, recombinant human TSLP was purchased from R&D Systems (Minneapolis, MN, USA) and BD Cytofix/Cytoperm and Alexa Fluor 647 mouse anti-STAT5 (pY694) were purchased from BD Biosciences (San Diego, CA, USA).

Techniques: Isolation

Journal: Molecules

Article Title: Lignans from Machilus thunbergii as Thymic Stromal Lymphopoietin Inhibitors

doi: 10.3390/molecules26164804

Figure Lengend Snippet: Molecular docking results of isolated compounds 1 – 3 against human TSLP in complex with TSLPR and interleukin-7 receptor α chain (IL-7Rα): ( A ) three-dimensional (3D) docking image of compound 1 ; ( B ) two-dimensional (2D) docking image of compound 1 ; ( C ) 3D docking image of compound 2 ; ( D ) 2D image picture of compound 2 ; ( E ) 3D docking image of compound 3 ; ( F ) 2D docking picture of compound 3 .

Article Snippet: For the pSTAT5 assay measuring TSLP inhibitory activity, recombinant human TSLP was purchased from R&D Systems (Minneapolis, MN, USA) and BD Cytofix/Cytoperm and Alexa Fluor 647 mouse anti-STAT5 (pY694) were purchased from BD Biosciences (San Diego, CA, USA).

Techniques: Isolation

Journal: Journal of clinical medicine

Article Title: TSLP Exacerbates Septic Inflammation via Murine Double Minute 2 (MDM2) Signaling Pathway.

doi: 10.3390/jcm8091350

Figure Lengend Snippet: Figure 1. TSLP is associated with sepsis. (A) TSLP levels in serum of patients with sepsis were analyzed by ELISA. Normal (Healthy volunteers, n = 20); Sepsis (Patients with sepsis, n = 30). A p value indicates the significant difference between normal and sepsis (B) TSLP levels were analyzed in serum of mice following LPS or E. coli injection by ELISA. (C) TSLP levels were analyzed in peritoneal lavage of mice following LPS or E. coli injection by ELISA. (D) TSLP protein levels were analyzed 12 h after LPS (10 mg/kg) or E. coli (1 × 106 CFU) injection by ELISA. (E) TSLP mRNA expression was analyzed 12 h after LPS (10 mg/kg) or E. coli (1 × 106 CFU) injection by real-time PCR (n = 10/group). A p value indicates the significant difference between PBS and LPS. PBS, phosphate-buffered saline; LPS, lipopolysaccharide; TSLP, thymic stromal lymphopoietin.

Article Snippet: Mice were intravenously injected with recombinant mouse TSLP (R&D Systems, 2 μg mixed with PBS) or PBS as a control (Sigma-Aldrich Co.) by referring to the studies of Piliponsky et al. [10] and Guo et al. [20].

Techniques: Enzyme-linked Immunosorbent Assay, Injection, Expressing, Real-time Polymerase Chain Reaction, Saline

Journal: Journal of clinical medicine

Article Title: TSLP Exacerbates Septic Inflammation via Murine Double Minute 2 (MDM2) Signaling Pathway.

doi: 10.3390/jcm8091350

Figure Lengend Snippet: Figure 2. Systemic inflammatory reaction is blunted by the deficiency of TSLP during sepsis. Mice were given either scramble control or TSLP-specific siRNA + atelocollagen mixture. Each level in (A) serum at 4 h and (B) liver homogenate at 12 h following LPS (10 mg/kg) injection was analyzed by ELISA. Adducts were normalized to total protein in liver homogenate. (C) Each level was analyzed in serum (n = 10/group). A p value indicates the significant difference between PBS and LPS. ** p < 0.05 vs. Con siRNA-received and LPS-injected control mice. PBS, phosphate-buffered saline; LPS, lipopolysaccharide; TSLP, thymic stromal lymphopoietin; Con, control; siRNA, small interfering RNA; VEGF, vascular endothelial growth factor; ICAM-1, intercellular adhesion molecule-1; MIP2, macrophage inflammatory protein 2; AST, aspartate aminotransferase; ALT, alanine aminotransferase, BUN, blood urea nitrogen; CK, creatine kinase.

Article Snippet: Mice were intravenously injected with recombinant mouse TSLP (R&D Systems, 2 μg mixed with PBS) or PBS as a control (Sigma-Aldrich Co.) by referring to the studies of Piliponsky et al. [10] and Guo et al. [20].

Techniques: Control, Injection, Enzyme-linked Immunosorbent Assay, Saline, Small Interfering RNA

Journal: Journal of clinical medicine

Article Title: TSLP Exacerbates Septic Inflammation via Murine Double Minute 2 (MDM2) Signaling Pathway.

doi: 10.3390/jcm8091350

Figure Lengend Snippet: Figure 3. TSLP is produced in macrophages. (A) RAW 264.7 cells were stimulated with LPS or E. coli for 24 h. (A, upper panel) Cell viability was analyzed by an MTT assay. (A, lower panel) NO concentration was measured by the Griess method. (B,C) Cells were stimulated with LPS or E. coli (upper panel) for 24 h for ELISA or (lower panel) 8 h for real-time PCR. Data are representative of three independent experiments (n = 5/group). A p value indicates the significant difference between PBS and LPS. LPS, lipopolysaccharide; TSLP, thymic stromal lymphopoietin; NO, nitric oxide.

Article Snippet: Mice were intravenously injected with recombinant mouse TSLP (R&D Systems, 2 μg mixed with PBS) or PBS as a control (Sigma-Aldrich Co.) by referring to the studies of Piliponsky et al. [10] and Guo et al. [20].

Techniques: Produced, MTT Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction

Journal: Journal of clinical medicine

Article Title: TSLP Exacerbates Septic Inflammation via Murine Double Minute 2 (MDM2) Signaling Pathway.

doi: 10.3390/jcm8091350

Figure Lengend Snippet: Figure 4. TSLP is produced via MDM2 signaling in macrophages. (A,B upper panel) The mRNA and (A,B lower panel) protein expressions of MDM2 were analyzed by real-time PCR and Western blot. (C, upper panel) The mRNA and (C, lower panel) protein expressions of MDM2 in liver were analyzed by real-time PCR and Western blot. (D, left) The production of TSLP 24 h and (D, right) mRNA expression of TSLP 8 h after LPS stimulation were analyzed in RAW 264.7 cells by ELISA and real-time PCR. ** p < 0.05 vs. Con siRNA transfection and LPS stimulation. (E) Cell viability was analyzed by an MTT assay. RAW 264.7 cells were treated with nutlin-3a for 2 h and stimulated with LPS (F, left) for 24 h for ELISA and (F, right) 8 h for real-time PCR. Data are representative of three independent experiments (n = 5/group). # p < 0.05 vs. LPS stimulation. (G) Serum at 4 h and (H) liver homogenate at 12 h following LPS injection from nutlin-3a-treated septic mice were subjected to ELISA (n = 10/group). Adducts were normalized to total protein in liver homogenate. 0.001% DMSO was treated as a vehicle negative control for nutlin-3a in LPS-unstimulated group. A p value indicates the significant difference between PBS and LPS. # p < 0.05 vs. LPS-injected mice. PBS, phosphate-buffered saline; LPS, lipopolysaccharide; TSLP, thymic stromal lymphopoietin; Con, control; siRNA, small interfering RNA; MDM2, murine double minute 2.

Article Snippet: Mice were intravenously injected with recombinant mouse TSLP (R&D Systems, 2 μg mixed with PBS) or PBS as a control (Sigma-Aldrich Co.) by referring to the studies of Piliponsky et al. [10] and Guo et al. [20].

Techniques: Produced, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Transfection, MTT Assay, Injection, Negative Control, Saline, Control, Small Interfering RNA

Journal: Journal of clinical medicine

Article Title: TSLP Exacerbates Septic Inflammation via Murine Double Minute 2 (MDM2) Signaling Pathway.

doi: 10.3390/jcm8091350

Figure Lengend Snippet: Figure 5. TSLP-dependent proinflammatory environment predisposes to the development of sepsis. (A) The mRNA expression of TSLPR and IL-7Rα were analyzed in RAW 264.7 cells by real-time PCR. (B) RAW 264.7 cells were stimulated with LPS (0.1 µg/mL) in the absence or presence of neutralizing anti-TSLP antibody (5 µg/mL) or nonimmune IgG (5 µg/mL) for 24 h. # p < 0.05 vs. LPS stimulation. RAW 264.7 cells were transfected with scramble control siRNA or TSLP-specific siRNA. After LPS stimulation, the mRNA expression levels of (C) TSLP, (D) TSLPR, and IL-7Rα were analyzed in the transfected cells by real-time PCR. (E) The transfected RAW 264.7 cells were stimulated with LPS for 24 h. ** p < 0.05 vs. Con siRNA transfection and LPS stimulation. (F) The peritoneal macrophages isolated from TSLP−/−

Article Snippet: Mice were intravenously injected with recombinant mouse TSLP (R&D Systems, 2 μg mixed with PBS) or PBS as a control (Sigma-Aldrich Co.) by referring to the studies of Piliponsky et al. [10] and Guo et al. [20].

Techniques: Expressing, Real-time Polymerase Chain Reaction, Transfection, Control, Isolation

Journal: Journal of clinical medicine

Article Title: TSLP Exacerbates Septic Inflammation via Murine Double Minute 2 (MDM2) Signaling Pathway.

doi: 10.3390/jcm8091350

Figure Lengend Snippet: Figure 6. Pharmacological inhibition of TSLP by cisplatin protects mice against lethal sepsis. (A) RAW 264.7 cells were treated with cisplatin for 2 h, followed by incubation for 24 h with LPS (0.1 µg/mL). Cell viability was analyzed by MTT assay. RAW 264.7 cells were treated with cisplatin for 2 h, followed by incubation for (B, Left) 24 h with LPS (0.1 µg/mL) for analysis of TSLP production and (B, Right) for 8 h with LPS (0.1 µg/mL) for analysis of TSLP mRNA expression. # p < 0.05 vs. LPS stimulation. (C) RAW 264.7 cells were treated with cisplatin for 2 h, followed by incubation for 8 h with LPS (0.1 µg/mL). MDM2 expression in cells lysates was analyzed by Western blot. Each TSLP level in (D) serum at 4 h and (E) liver homogenate at 12 h following LPS (10 mg/kg) injection was analyzed by ELISA. Adducts were normalized to total protein in liver homogenate. (F) MDM2 expression in liver homogenate at 12 h following LPS (10 mg/kg) injection was analyzed by Western blot. For immunoblots, GAPDH was used as a loading control. (G) BUN, CK, and AST levels were analyzed in the serum at 4 h following LPS (10 mg/kg) injection (n = 10/group). PBS was treated as a vehicle negative control for cisplatin in LPS-unstimulated group. A p value indicates the significant difference between PBS and LPS. # p < 0.05 vs. LPS stimulation. # p < 0.05 vs. LPS-injected mice. (H) Survival curve (%) was monitored in mice (n = 10/group) injected intraperitoneally with LPS (60 mg/kg), E. coli (1 × 108 CFU), or TSLP (2 µg) following intravenous injection of cisplatin (100 µg/kg). LPS, lipopolysaccharide; TSLP, thymic stromal lymphopoietin; MDM2, murine double minute 2; BUN, blood urea nitrogen; CK, creatine kinase; AST, aspartate aminotransferase.

Article Snippet: Mice were intravenously injected with recombinant mouse TSLP (R&D Systems, 2 μg mixed with PBS) or PBS as a control (Sigma-Aldrich Co.) by referring to the studies of Piliponsky et al. [10] and Guo et al. [20].

Techniques: Inhibition, Incubation, MTT Assay, Expressing, Western Blot, Injection, Enzyme-linked Immunosorbent Assay, Control, Negative Control

Journal: Journal of clinical medicine

Article Title: TSLP Exacerbates Septic Inflammation via Murine Double Minute 2 (MDM2) Signaling Pathway.

doi: 10.3390/jcm8091350

Figure Lengend Snippet: Figure 7. Graphical abstract of TSLP function during sepsis. Production of TSLP is induced via the MDM2/NF-κB signaling pathway in E. coli-stimulated macrophages. TSLP affects inflammatory cytokines production and organ dysfunction during sepsis. Cisplatin protects mice against lethal sepsis by inhibiting the series of reactions. TSLP can be induced directly by E. coli or other proinflammatory stimuli. Multiple cell types such as neutrophils, dendritic cells, natural killer cells, and T cells might be involved in sepsis.

Article Snippet: Mice were intravenously injected with recombinant mouse TSLP (R&D Systems, 2 μg mixed with PBS) or PBS as a control (Sigma-Aldrich Co.) by referring to the studies of Piliponsky et al. [10] and Guo et al. [20].

Techniques:

Journal: iScience

Article Title: Immunometabolic adaptation in monocytes underpins functional changes during pregnancy

doi: 10.1016/j.isci.2024.109779

Figure Lengend Snippet:

Article Snippet: CD163 human antibody (clone GHI/61.1) , Miltenyi , Cat#130-123-249; RRID: AB_2819455.

Techniques: Recombinant, Adhesive, Sequencing, Modification, DC Protein Assay, Staining, Imaging, Enzyme-linked Immunosorbent Assay, Bicinchoninic Acid Protein Assay, Gene Expression, Software, Saline