Journal: Frontiers in cell and developmental biology

Article Title: Lysophosphatidylcholine Alleviates Acute Lung Injury by Regulating Neutrophil Motility and Neutrophil Extracellular Trap Formation.

doi: 10.3389/fcell.2022.941914

Figure Lengend Snippet: FIGURE 1 | LPC obstructs mouse neutrophil migration during LPS stimulation. (A,B) Migration of mouse neutrophils on a fibronectin-coated confocal dish in the presence or absence of LPC under inflammatory conditions. Given conditions were treated with LPS (0.1 μg/ml) and LPS + LPC (LPS 0.1 μg/ml co-treated with 30 µM LPC) and incubated for 1 h on serum-starved neutrophils. Length unit, μm. (C) Velocity, (D) Displacement, and (E) Meandering index. (F) Mean fluorescence intensity of CD11a and (G) CD11b adhesion molecules on neutrophils. All experiments were independently repeated at least three times. ***p < 0.001, **p < 0.01, *p < 0.05.

Article Snippet: Two hours after LPS inoculation, 18:0 LPC (10 mg/kg; Avanti Polar Lipids, Birmingham, AL, United States) was subcutaneously injected at different sites four times at 12 h intervals for 48 h. All animal experiments were approved by the Institutional Animal Care and Use Committee of the Yonsei University College of Medicine (IACUC No. 2019–0,097).

Techniques: Migration, Incubation

Journal: Frontiers in cell and developmental biology

Article Title: Lysophosphatidylcholine Alleviates Acute Lung Injury by Regulating Neutrophil Motility and Neutrophil Extracellular Trap Formation.

doi: 10.3389/fcell.2022.941914

Figure Lengend Snippet: FIGURE 3 | LPC reduces the formation of mouse neutrophil extracellular trap (NET). (A) Fluorescent images showing NET formation. Representative images were acquired by staining for Sytox-orange (red), CitH3 (green) and nuclei (blue). (B) Western blots of MPO and CitH3; β-actin was used as a control. (C,D) The expression of MPO and CitH3 was measured by Western blot in lung tissues of LPS group and LPS + LPC group. Results were normalized to loading control (β-actin). (E) NET localization in the lung of a mouse with LPS-induced sepsis. Immunofluorescence staining of lung sections and NET formation. MPO (green) and CitH3 (red) labeling were used to detect NET release. Nuclei were stained with DAPI (blue). White lines indicate examples of double-stained NETs. (F) Representative images of ROS release (green) in mouse neutrophils. Scale bars, 30 μm. A representative data was shown from at least three times independent experiments.

Article Snippet: Two hours after LPS inoculation, 18:0 LPC (10 mg/kg; Avanti Polar Lipids, Birmingham, AL, United States) was subcutaneously injected at different sites four times at 12 h intervals for 48 h. All animal experiments were approved by the Institutional Animal Care and Use Committee of the Yonsei University College of Medicine (IACUC No. 2019–0,097).

Techniques: Staining, Western Blot, Control, Expressing, Labeling

Journal: Frontiers in cell and developmental biology

Article Title: Lysophosphatidylcholine Alleviates Acute Lung Injury by Regulating Neutrophil Motility and Neutrophil Extracellular Trap Formation.

doi: 10.3389/fcell.2022.941914

Figure Lengend Snippet: FIGURE 5 | LPC reduces lung tissue damage and decreased innate immune cell infiltration in the inflammatory lung. (A) Schematic diagram of in vivo experiment (B) Mouse lung was imaged to identify neutrophils (green) and bloodstream (red). Scale bar, 50 µm. (C) Clustered neutrophils were less abundant in LPC-treated mice. (D) Confirmation of immune cells in inflamed lungs by flow cytometry. Innate immune cells were sorted using CD11b, Ly6G, and F4/80. (E) Quantitative confirmation by flow cytometry. (F) Representative images showing H&E staining in LPS-inflamed lung tissue for 48 h with and without LPC administration. (G) Histological score of lung injury. A representative data was shown from three times independent experiments. **p < 0.01 compared with LPS-only group.

Article Snippet: Two hours after LPS inoculation, 18:0 LPC (10 mg/kg; Avanti Polar Lipids, Birmingham, AL, United States) was subcutaneously injected at different sites four times at 12 h intervals for 48 h. All animal experiments were approved by the Institutional Animal Care and Use Committee of the Yonsei University College of Medicine (IACUC No. 2019–0,097).

Techniques: In Vivo, Cytometry, Staining

Journal: Frontiers in cell and developmental biology

Article Title: Lysophosphatidylcholine Alleviates Acute Lung Injury by Regulating Neutrophil Motility and Neutrophil Extracellular Trap Formation.

doi: 10.3389/fcell.2022.941914

Figure Lengend Snippet: FIGURE 6 | LPC restores the adhesion process inhibited by the Mac-1 blockade. (A) Neutrophil migration was confirmed in the human blood neutrophils of a healthy donor stimulated with LPS (1 μg/ml) and concurrently treated with LPC (30 µM). Adhesion of neutrophils via Mac-1 blockade was also confirmed. Scale bars, 50 μm, Length unit, μm. (B) Velocity, (C) displacement, and (D) meandering index were checked according to each LPS and LPC treatment group. **p < 0.005, *p < 0.05. A representative data was shown from three times independent experiments.

Article Snippet: Two hours after LPS inoculation, 18:0 LPC (10 mg/kg; Avanti Polar Lipids, Birmingham, AL, United States) was subcutaneously injected at different sites four times at 12 h intervals for 48 h. All animal experiments were approved by the Institutional Animal Care and Use Committee of the Yonsei University College of Medicine (IACUC No. 2019–0,097).

Techniques: Migration