Journal: Aging

Article Title: Roles of pigment epithelium-derived factor in exercise-induced suppression of senescence and its impact on lung pathology in mice.

doi: 10.18632/aging.205976

Figure Lengend Snippet: Figure 2. PEDF mediates the anti-cellular senescence effects of C2C12-CM. (A–C) MEFs were cultured in the presence of C2C12-CM treated with a control or PEDF antibody for 3 days. (A) Cell numbers were counted and relative changes in cell numbers in 3 days were plotted. (B) Total RNA was isolated from MEFs and the expression of Ink4a and Arf was analyzed by real-time PCR. Values were normalized to Gapdh in each sample. (C) p16INK4a and p19ARF levels were analyzed by immunoblotting. β-Actin was used as the loading control. (D) MEFs were cultured in the presence of a recombinant of PEDF (100 ng/mL) for 3 days. Changes in cell numbers were plotted. (E) Cell viability was determined by the trypan blue exclusion assay. (F) The expression of Ink4a, Arf, and p21 was analyzed by real-time PCR. Values were normalized to Gapdh in each sample. (G) p16INK4a, p19ARF, and p21 levels were analyzed by immunoblotting. β-Actin was used as a loading control. (H) Cells were stained for SA-β-gal. Scale bar, 100 μm. (I) The percentage of SA-β-gal-positive cells was plotted. (J) Cells were stimulated with the indicated concentrations of recombinant PEDF for 3 days. Intracellular ROS levels were analyzed in each sample, and relative values were plotted against the average of the control sample. Values represent means ± SD. Data were analyzed by the Student’s t-test (A, B, D–F, I) or a one-way ANOVA and Tukey’s post-hoc analysis (J). *P <0.05, **P <0.01, and ***P <0.001.

Article Snippet: Samples were analyzed using a Mouse PEDF, PEDF ELISA Kit (CSB-E08820m, Cusabio Technology, LLC).

Techniques: Cell Culture, Control, Isolation, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Recombinant, Trypan Blue Exclusion Assay, Staining

Journal: Aging

Article Title: Roles of pigment epithelium-derived factor in exercise-induced suppression of senescence and its impact on lung pathology in mice.

doi: 10.18632/aging.205976

Figure Lengend Snippet: Figure 3. Voluntary exercise up-regulates PEDF and suppresses cellular senescence. Six-month-old wild-type mice were subjected to voluntary exercise for 8 weeks. (A) Total RNA was isolated from skeletal muscles (TA and SOL) and Pedf levels were analyzed by real-time PCR. Values were normalized to 18S rRNA in each sample. (B) Serum PEDF levels were analyzed by ELISA. (C) The expression of Ink4a, Arf and p21 in the lungs was analyzed by real-time PCR. Values were normalized to Gapdh in each sample. (D) Lung sections were stained for SA-β- gal. Arrowheads indicate SA-β-gal-positive cells. Scale bar, 100 μm. (E) The number of SA-β-gal-positive cells was counted in each sample. (F) The expression of Il-1b, Il-6, and Mmp-12 in lung total RNA was analyzed by real-time PCR. Values were normalized to 18S-rRNA in each sample. Values represent means ± SEM. Data were analyzed by the Student’s t-test. *P <0.05, **P <0.01, and ***P <0.001.

Article Snippet: Samples were analyzed using a Mouse PEDF, PEDF ELISA Kit (CSB-E08820m, Cusabio Technology, LLC).

Techniques: Isolation, Muscles, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Expressing, Staining

Journal: Aging

Article Title: Roles of pigment epithelium-derived factor in exercise-induced suppression of senescence and its impact on lung pathology in mice.

doi: 10.18632/aging.205976

Figure Lengend Snippet: Figure 4. PEDF suppresses cellular senescence in lung tissues. A recombinant PEDF protein (10 μg/kg body weight) was intraperitoneally injected twice a week for 4 weeks. (A) The expression of Ink4a, Arf, and p21 in lung total RNA was analyzed by real-time PCR. Values were normalized to Gapdh in each sample. (B) Lung tissues were stained for SA-β-gal. Arrowheads indicate SA-β-gal-positive cells. Scale bar, 50 μm. (C) The number of SA-β-gal-positive cells was counted in each sample. Values represent means ± SEM. Data were analyzed by the Student’s t-test. *P <0.05 and **P <0.01.

Article Snippet: Samples were analyzed using a Mouse PEDF, PEDF ELISA Kit (CSB-E08820m, Cusabio Technology, LLC).

Techniques: Recombinant, Injection, Expressing, Real-time Polymerase Chain Reaction, Staining

Journal: Aging

Article Title: Roles of pigment epithelium-derived factor in exercise-induced suppression of senescence and its impact on lung pathology in mice.

doi: 10.18632/aging.205976

Figure Lengend Snippet: Figure 5. PEDF protected lung tissues from PPE-induced emphysema. (A) Experimental design. A recombinant PEDF protein was injected intraperitoneally twice a week for 4 weeks. PPE (5 units) was intranasally administered 2 weeks after the first dose of PEDF. Mice were analyzed 3 weeks after the PPE treatment. (B) Total RNA was isolated from lung tissues and the expression of Ink4a, Arf, and p21 was analyzed by real-time PCR. Values were normalized to Gapdh in each sample. (C) Representative images of control and PEDF-treated mouse lung sections. Sections were stained with hematoxylin and eosin. Scale bar, 100 μm. (D) Alveolar mean linear intercepts were measured. (E–H) A pulmonary function test was performed. Lung compliance (E), capacity (F), tissue damping (G), and tissue elastance (H) are shown. Values represent means ± SEM. Data were analyzed by the Student’s t-test (B), or a one-way ANOVA and Tukey’s post-hoc analysis (D–H). *P <0.05, **P <0.01, and ***P <0.001.

Article Snippet: Samples were analyzed using a Mouse PEDF, PEDF ELISA Kit (CSB-E08820m, Cusabio Technology, LLC).

Techniques: Recombinant, Injection, Isolation, Expressing, Real-time Polymerase Chain Reaction, Control, Staining

Journal: Aging

Article Title: Roles of pigment epithelium-derived factor in exercise-induced suppression of senescence and its impact on lung pathology in mice.

doi: 10.18632/aging.205976

Figure Lengend Snippet: Figure 6. Involvement of miR-127 in PEDF signaling. (A) An RNA sequencing analysis was performed on MEFs stimulated with a recombinant PEDF protein. A volcano plot of RNA sequencing data is shown. A full list of RNA sequencing data is available in the Gene Expression Omnibus database (accession number; GSE241459). (B) The expression of miR-127 (-loop, -5p, and -3p) in MEFs was analyzed by real-time PCR. Values were normalized to U6 snRNA in each sample. (C) The expression of BCL-6 in MEFs was analyzed by immunoblotting. β- Actin was used as a loading control. (D, E) miR-127 levels in the lungs of mice subjected to the PEDF treatment (D, Figure 4) or voluntary wheel running (E, Figure 3) were analyzed by real-time PCR. miR-127 levels were normalized to U6 snRNA in each sample. Values represent means ± SD (B) or means ± SEM (D, E). Data were analyzed by the Student’s t-test. *P <0.05, **P <0.01, and ***P <0.001.

Article Snippet: Samples were analyzed using a Mouse PEDF, PEDF ELISA Kit (CSB-E08820m, Cusabio Technology, LLC).

Techniques: RNA Sequencing, Recombinant, Gene Expression, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: PLoS ONE

Article Title: Diosmin Alleviates Retinal Edema by Protecting the Blood-Retinal Barrier and Reducing Retinal Vascular Permeability during Ischemia/Reperfusion Injury

doi: 10.1371/journal.pone.0061794

Figure Lengend Snippet: (A) Twenty-four hours after ischemia, there was a significant increase in VEGF expression and a slight increase in PEDF expression in the vehicle group compared with the control group; diosmin treatment reversed this trend. (B) Seven days after ischemia, the VEGF and PEDF protein expression returned to normal in both the vehicle and diosmin groups. (C) The VEGF/PEDF ratio for the three groups 24 h and 7 d after ischemia. Data are expressed as the mean±SD ( n = 8). ** P <0.01, *** P <0.001.

Article Snippet: In each sample, the concentrations of VEGF (Rat VEGF Quantikine ELISA Kit; R&D Systems, Minneapolis, MN) and PEDF (Rat pigment epithelium-derived factor, PEDF ELISA Kit; EIAab ®, Wuhan, China) were measured with ELISA kits according to the respective manufacturer’s protocol.

Techniques: Expressing, Control

Journal: PLoS ONE

Article Title: Diosmin Alleviates Retinal Edema by Protecting the Blood-Retinal Barrier and Reducing Retinal Vascular Permeability during Ischemia/Reperfusion Injury

doi: 10.1371/journal.pone.0061794

Figure Lengend Snippet: No significant immunoreactivity for VEGF and PEDF was found in control retina. I/R injury induced a stronger immunoreactivity for VEGF in retina and diosmin administration relieved this trend. The PEDF immunoreactivity was nearly similar to the control group after 24 h retinal ischemia, while diosmin administration immensely increased this immunoreactivity. Scale bar, 100 µm.

Article Snippet: In each sample, the concentrations of VEGF (Rat VEGF Quantikine ELISA Kit; R&D Systems, Minneapolis, MN) and PEDF (Rat pigment epithelium-derived factor, PEDF ELISA Kit; EIAab ®, Wuhan, China) were measured with ELISA kits according to the respective manufacturer’s protocol.

Techniques: Control

Journal: Medicina

Article Title: Changes in Plasma VEGF and PEDF Levels in Patients with Central Serous Chorioretinopathy

doi: 10.3390/medicina57101063

Figure Lengend Snippet: Plasma levels of vascular endothelial growth factor (VEGF) and pigment-epithelium-derived factor (PEDF) in patients with acute central serous chorioretinopathy (CSC), patients with chronic CSC, and controls.

Article Snippet: Finally, PEDF levels were measured at a plasma dilution of 1:2 using PEDF ELISA Kit (Biomatik, Wilmington, DE, USA) according to the manufacturer’s protocol.

Techniques: Clinical Proteomics

Journal: Medicina

Article Title: Changes in Plasma VEGF and PEDF Levels in Patients with Central Serous Chorioretinopathy

doi: 10.3390/medicina57101063

Figure Lengend Snippet: Box-and-whisker plot of the ratio of VEGF to pigment-epithelium–derived factor (PEDF) in all patients with central serous chorioretinopathy and controls ( p = 0.04).

Article Snippet: Finally, PEDF levels were measured at a plasma dilution of 1:2 using PEDF ELISA Kit (Biomatik, Wilmington, DE, USA) according to the manufacturer’s protocol.

Techniques: Whisker Assay, Derivative Assay

Journal: Medicina

Article Title: Changes in Plasma VEGF and PEDF Levels in Patients with Central Serous Chorioretinopathy

doi: 10.3390/medicina57101063

Figure Lengend Snippet: Correlation between the plasma levels of PEDF and disease duration in patients with chronic CSC.

Article Snippet: Finally, PEDF levels were measured at a plasma dilution of 1:2 using PEDF ELISA Kit (Biomatik, Wilmington, DE, USA) according to the manufacturer’s protocol.

Techniques: Clinical Proteomics