Journal: Biomolecules

Article Title: Extracellular Vesicles Induce Nuclear Factor-κB Activation and Interleukin-8 Synthesis through miRNA-191-5p Contributing to Inflammatory Processes: Potential Implications in the Pathogenesis of Chronic Obstructive Pulmonary Disease

doi: 10.3390/biom14081030

Figure Lengend Snippet: Graphical overview of the experimental design. Bronchial epithelial cells (16HBE) were transfected with miR-191-5p mimic or inhibitor. Subsequently, supernatants were collected for EV isolation and miR-191-5p quantification. Additionally, cells were harvested for miRNA extraction. The bronchial epithelial cells were then exposed to EVs (EV m : EV derived from 16HBE transfected with miR-191-5p mimic; EV i : EV derived from 16HBE transfected with miR-191-5p inhibitor; EV c : EV derived from 16HBE transfected with miRNA scramble; EV nt : EV derived from 16HBE not transfected) and subsequently collected for miR-191-5p quantification, NF-kκb activation and IL-8 evaluation. The figure was created through “biorender.com” website license agreement number EX2679WEP4, access date 11 December 2024.

Article Snippet: miRNA mimic of miR-191-5p (m-miR-191-5p) and miR-191-5p (i-miR-191-5p) inhibitor were purchased from Qiagen (Hilden, Germany).

Techniques: Transfection, Isolation, Extraction, Derivative Assay, Activation Assay

Journal: Biomolecules

Article Title: Extracellular Vesicles Induce Nuclear Factor-κB Activation and Interleukin-8 Synthesis through miRNA-191-5p Contributing to Inflammatory Processes: Potential Implications in the Pathogenesis of Chronic Obstructive Pulmonary Disease

doi: 10.3390/biom14081030

Figure Lengend Snippet: Expression of miR-191-5p in 16HBE cells ( A ) and in EVs ( B ) after transfection with miRNA mimic of miR-191-5p (mimic-miR-191-5p) and miR-191-5p inhibitor (inhibitor -miR-191-5p). The mimic scramble was used as control (Ctrl) in each experiment. Non-transfected cells represent the not-treated group (Untreated). Expression of miR-191-5p in 16HBE cells ( C ) after incubation with EVs shed from bronchial epithelial cells transfected with mimic-miR-191-5p and inhibitor-miR-191-5p (EV m : EV derived from 16HBE transfected with miR-191-5p mimic; EV i : EV derived from 16HBE transfected with miR-191-5p inhibitor; EV c : EV derived from 16HBE transfected with miRNA scramble; EV nt : EV derived from 16HBE not transfected). Statistical analysis was performed using one-way ANOVA followed by Bonferroni’s test selected data (* p < 0.05 vs. Ctrl).

Article Snippet: miRNA mimic of miR-191-5p (m-miR-191-5p) and miR-191-5p (i-miR-191-5p) inhibitor were purchased from Qiagen (Hilden, Germany).

Techniques: Expressing, Transfection, Control, Incubation, Derivative Assay

Journal: Biomolecules

Article Title: Extracellular Vesicles Induce Nuclear Factor-κB Activation and Interleukin-8 Synthesis through miRNA-191-5p Contributing to Inflammatory Processes: Potential Implications in the Pathogenesis of Chronic Obstructive Pulmonary Disease

doi: 10.3390/biom14081030

Figure Lengend Snippet: Modulation of NF-κB and IL-8 by miR-191-5p-enirched EVs. ( A ) Activation of NF-κB in 16HBE cells exposed to EV derived from Ach-stimulated 16HBE cells treated as described in A (EV m : EV derived from 16HBE transfected with miR-191-5p mimic; EV i : EV derived from 16HBE transfected with miR-191-5p inhibitor; EV c : EV derived from 16HBE transfected with miRNA scramble; EV nt : EV derived from 16HBE not transfected). Then, nuclear proteins were extracted and assessed by ELISA to measure the DNA-binding activity of the p65 NF-κB subunit. ( B ) IL-8 secretion by 16HBE cells incubated with EVs derived from Ach-stimulated 16HBE cells, treated as described in A. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparison test (*** p < 0.001 vs. 16HBE + EV c ).

Article Snippet: miRNA mimic of miR-191-5p (m-miR-191-5p) and miR-191-5p (i-miR-191-5p) inhibitor were purchased from Qiagen (Hilden, Germany).

Techniques: Activation Assay, Derivative Assay, Transfection, Enzyme-linked Immunosorbent Assay, Binding Assay, Activity Assay, Incubation, Comparison

Journal: Biomolecules

Article Title: Extracellular Vesicles Induce Nuclear Factor-κB Activation and Interleukin-8 Synthesis through miRNA-191-5p Contributing to Inflammatory Processes: Potential Implications in the Pathogenesis of Chronic Obstructive Pulmonary Disease

doi: 10.3390/biom14081030

Figure Lengend Snippet: Correlation of EV-derived miR-191-5p and clinical parameters. Correlation analysis was performed to assess the relationships between EV-derived miR-191-5p levels and specific clinical parameters, including IL-6 ( A ) and DLCO ( B ). For technical reasons, IL-6 level was not available in one patient. The analysis was conducted using simple linear regression analysis.

Article Snippet: miRNA mimic of miR-191-5p (m-miR-191-5p) and miR-191-5p (i-miR-191-5p) inhibitor were purchased from Qiagen (Hilden, Germany).

Techniques: Derivative Assay

Journal: Biomolecules

Article Title: Extracellular Vesicles Induce Nuclear Factor-κB Activation and Interleukin-8 Synthesis through miRNA-191-5p Contributing to Inflammatory Processes: Potential Implications in the Pathogenesis of Chronic Obstructive Pulmonary Disease

doi: 10.3390/biom14081030

Figure Lengend Snippet: Predictive-network-based analysis involving the activation of NF-κB ( A ), IL-6 ( B ), and IL-8 ( C ). The modulation of the three targets predicted a positive correlation with miR-191-5p, as well as with other nodes related to disease, including IL-1, CRP, and TNF-α, among others. The bioinformatic survey pinpointed a signaling pathway consistent with expression results and reconstructed the molecular neighborhood around targets with several connection nodes that are valuable for a more comprehensive evaluation of the role of miR-191-5p in the pathogenesis of COPD.

Article Snippet: miRNA mimic of miR-191-5p (m-miR-191-5p) and miR-191-5p (i-miR-191-5p) inhibitor were purchased from Qiagen (Hilden, Germany).

Techniques: Activation Assay, Expressing