Journal: Molecular Medicine Reports

Article Title: High‑fat diet‑induced LCN2 exacerbates myocardial ischemia‑reperfusion injury by enhancing platelet activation

doi: 10.3892/mmr.2024.13329

Figure Lengend Snippet: Expression of LCN2 in the liver and serum of HFD-fed mice. (A) Body weight was measured in mice that were fed a ND or HFD. (B) Liver tissue was analyzed using hematoxylin and eosin staining to observe pathological changes caused by a HFD. Lipid droplets were indicated by black arrows. (C) Reverse transcription-quantitative PCR was performed to assess the expression levels of LCN2 in the livers of mice who were or were not fed a HFD. (D) Levels of LCN2 in the serum of mice who were or were not fed a HFD were measured by ELISA. Data are presented as the mean ± SEM (n=5 mice/group). *P<0.05. HFD, high-fat diet; LCN2, lipocalin 2; ND, normal diet.

Article Snippet: Platelets were exposed to LCN2 (2 μg/ml; cat. no. 1757-LC-050; R&D Systems, Inc.) for 5 min at 37°C, and were then stimulated with thrombin (0.04 U/ml; cat. no. T6884; Sigma-Aldrich; Merck KGaA), collagen (1 μg/ml; cat. no. P/N 385; Chrono-Log Corporation) or adenosine diphosphate (ADP; 2.5 μM; cat. no. A2754; Sigma-Aldrich; Merck KGaA) for 5 min at 37°C.

Techniques: Expressing, Staining, Reverse Transcription, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Molecular Medicine Reports

Article Title: High‑fat diet‑induced LCN2 exacerbates myocardial ischemia‑reperfusion injury by enhancing platelet activation

doi: 10.3892/mmr.2024.13329

Figure Lengend Snippet: Effect of LCN2 on platelet function. (A) After platelets were incubated with or without LCN2, they were further stimulated with ADP, thrombin or collagen to detect platelet aggregation (5 platelet samples were collected from 5 volunteers). (B) After platelets from different treatment groups were incubated on cellular coverslips for 45 min, they were stained with iFluor ™ 680-phalloidin, and the adhesion of platelets was observed under a fluorescence microscope (5 platelet samples were collected from 5 volunteers). (C) After incubation with or without LCN2, the platelets were stimulated with thrombin. Clot retraction was observed after stimulation with thrombin for 0, 20, 40 and 60 min (5 platelet samples were collected from 5 volunteers). (D) Upon stimulation with or without LCN2, the platelets were incubated with thrombin, and untreated platelets were used as a control. The expression of P-selectin on the platelet surface was detected by flow cytometry (3 platelet samples were collected from 3 volunteers). Data are presented as the mean ± SEM. *P<0.05. ADP, adenosine diphosphate; BSA, bovine serum albumin; LCN2, lipocalin 2.

Article Snippet: Platelets were exposed to LCN2 (2 μg/ml; cat. no. 1757-LC-050; R&D Systems, Inc.) for 5 min at 37°C, and were then stimulated with thrombin (0.04 U/ml; cat. no. T6884; Sigma-Aldrich; Merck KGaA), collagen (1 μg/ml; cat. no. P/N 385; Chrono-Log Corporation) or adenosine diphosphate (ADP; 2.5 μM; cat. no. A2754; Sigma-Aldrich; Merck KGaA) for 5 min at 37°C.

Techniques: Incubation, Staining, Fluorescence, Microscopy, Control, Expressing, Flow Cytometry

Journal: Molecular Medicine Reports

Article Title: High‑fat diet‑induced LCN2 exacerbates myocardial ischemia‑reperfusion injury by enhancing platelet activation

doi: 10.3892/mmr.2024.13329

Figure Lengend Snippet: Effect of LCN2 knockout on MI/R injury in HFD-fed mice. (A) Mouse tails were utilized for DNA extraction, followed by PCR amplification and electrophoresis. For WT mice, a PCR product of 1,287 bp was obtained; conversely, LCN2 −/− mice had a product of 586 bp. Subsequently, both WT and LCN2 −/− mice underwent MI/R injury and sham surgery after being fed a HFD. (B) Mouse hearts were procured for hematoxylin and eosin staining. (C) Degree of infarction was assessed through TTC staining. Data are presented as the mean ± SEM (n=5). *P<0.05. HFD, high-fat diet; LCN2, lipocalin 2; MI/R, myocardial ischemia-reperfusion; TTC, 2,3,5-triphenyltetrazolium chloride; WT, wild-type.

Article Snippet: Platelets were exposed to LCN2 (2 μg/ml; cat. no. 1757-LC-050; R&D Systems, Inc.) for 5 min at 37°C, and were then stimulated with thrombin (0.04 U/ml; cat. no. T6884; Sigma-Aldrich; Merck KGaA), collagen (1 μg/ml; cat. no. P/N 385; Chrono-Log Corporation) or adenosine diphosphate (ADP; 2.5 μM; cat. no. A2754; Sigma-Aldrich; Merck KGaA) for 5 min at 37°C.

Techniques: Knock-Out, DNA Extraction, Amplification, Electrophoresis, Staining

Journal: Molecular Medicine Reports

Article Title: High‑fat diet‑induced LCN2 exacerbates myocardial ischemia‑reperfusion injury by enhancing platelet activation

doi: 10.3892/mmr.2024.13329

Figure Lengend Snippet: Effect of LCN2 knockout on the recruitment of platelets and inflammatory cells in myocardial tissue after I/R injury in mice induced by a HFD. (A) WT and LCN2 −/− mice were administered a HFD, and were then subjected to MI/R injury. Immunohistochemistry was performed to assess the expression of CD42b in the collected hearts. (B) WT and LCN2 −/− mice were fed a HFD and were subjected to MI/R injury. Immunohistochemistry was then employed to detect the recruitment of Ly6G + , CD3 + and B220 + cells in myocardial tissues. Data are presented as the mean ± SEM (n=5). *P<0.05. HFD, high-fat diet; LCN2, lipocalin 2; MI/R, myocardial ischemia-reperfusion; WT, wild-type.

Article Snippet: Platelets were exposed to LCN2 (2 μg/ml; cat. no. 1757-LC-050; R&D Systems, Inc.) for 5 min at 37°C, and were then stimulated with thrombin (0.04 U/ml; cat. no. T6884; Sigma-Aldrich; Merck KGaA), collagen (1 μg/ml; cat. no. P/N 385; Chrono-Log Corporation) or adenosine diphosphate (ADP; 2.5 μM; cat. no. A2754; Sigma-Aldrich; Merck KGaA) for 5 min at 37°C.

Techniques: Knock-Out, Immunohistochemistry, Expressing