Journal: Molecular Medicine Reports

Article Title: Role of the Rho/ROCK signaling pathway in the protective effects of fasudil against acute lung injury in septic rats

doi: 10.3892/mmr.2018.9446

Figure Lengend Snippet: Effect of fasudil on the viability and the secretion of inflammatory cytokines from HPMVEC-ST1.6R treated with LPS. (A) The effect of LPS on the viability of HPMVEC-ST1.6R was examined by MTT assay. n=6. *P<0.05, **P<0.01, vs. control. (B) Fasudil relieved the cytotoxic effects of LPS on rat HPMVEC- ST1.6R examined by MTT assay. Data are expressed as the mean ± standard deviation. n=6. *P<0.05 vs. control group; # P<0.05 vs. LPS-only group. Fasudil inhibited LPS-induced (C) VEGF, (D) ICAM-1 and (E) VCAM-1 secretion from HPMVEC-ST1.6R. Fasudil (10, 25 and 50 µM) was separately preincubated with HPMVEC-ST1.6R 30 min prior to LPS exposure. Supernatants were detected using ELISA for VEGF, ICAM-1 and VCAM-1. Fasudil-L, fasudil at a concentration of 10 µM; fasudil-M, fasudil at a concentration of 25 µM; fasudil-H, fasudil at a concentration of 50 µM. Data are expressed as the mean ± standard deviation. n=6. # P<0.05 vs. LPS group. LPS, lipopolysaccharide, VEGF, vascular endothelial growth factor, ICAM-1, intracellular cell adhesion molecule-1; VCAM-1, vascular cell adhesion molecule 1; OD, optical density; HPMVEC, human pulmonary microvascular endothelial cells.

Article Snippet: Human pulmonary microvascular endothelial cells (HPMVEC-ST1.6R) were purchased from Clonetics TM (Lonza Group Ltd., Basel, Switzerland).

Techniques: MTT Assay, Control, Standard Deviation, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Molecular Medicine Reports

Article Title: Role of the Rho/ROCK signaling pathway in the protective effects of fasudil against acute lung injury in septic rats

doi: 10.3892/mmr.2018.9446

Figure Lengend Snippet: Effect of fasudil on the LPS-induced reorganization of actin filaments in HPMVEC-ST1.6R (magnification, ×600). HPMVEC-ST1.6R were pretreated with or without fasudil (50 µM) for 2 h, stimulated with LPS (10 µg/ml) for 24 h, and the F-actin stress fibers were stained with fluorescein isothiocyanate-conjugated phalloidin and visualized by confocal microscopy. Fasudil prevented the LPS-induced reorganization of actin filaments. LPS, lipopolysaccharide; HPMVEC, human pulmonary microvascular endothelial cells; F-actin, filamentous actin.

Article Snippet: Human pulmonary microvascular endothelial cells (HPMVEC-ST1.6R) were purchased from Clonetics TM (Lonza Group Ltd., Basel, Switzerland).

Techniques: Staining, Confocal Microscopy

Journal: Annals of Translational Medicine

Article Title: Sphingosine 1-phosphate receptor 1 governs endothelial barrier function and angiogenesis by upregulating endoglin signaling

doi: 10.21037/atm-21-6679

Figure Lengend Snippet: Activation and interaction of S1P and BMP9 downstream signaling in endothelial cells. (A) Confluent HPMVEC cells were treated with S1P (200 nM) or BMP9 (10 ng/mL) as indicated. Expressions of p-ERK42/44, total ERK42/44, p-Smad1/5/8, Smad1, and vinculin were analyzed using Western blot. Treatment with S1P induced phosphorylation of ERK proteins, whereas BMP9 only activated the phosphorylation of Smad1/5/8. (B,C) Expressions of Smad1 and p-Smad1/5/8 were analyzed using Western blot in HPMVEC cells (B) and MS1 cells (C) pretreated with BSA or S1P (200 nM) for 30 min followed with BMP9 (10 ng/mL) as indicated. Vinculin was used as a loading control. The right panels illustrate the ratio of p-Smad1/5/8 over Smad1 with histograms at each time point as indicated. Student t-test, **, P<0.01; ***, P<0.001. −, in the absence of S1P or BMP9; +, in the presence of S1P or BMP9. S1P, sphingosine 1-phosphate; BMP9, bone morphogenetic protein 9; HPMVEC, human pulmonary microvascular endothelial cell; ERK, extracellular regulated protein kinases; MS1, a mouse islet EC line; BSA, bovine serum albumin.

Article Snippet: HPMVEC (human pulmonary microvascular endothelial cell) was purchased from Lonza (Portsmouth, USA).

Techniques: Activation Assay, Western Blot, Phospho-proteomics, Control

Journal: Annals of Translational Medicine

Article Title: Sphingosine 1-phosphate receptor 1 governs endothelial barrier function and angiogenesis by upregulating endoglin signaling

doi: 10.21037/atm-21-6679

Figure Lengend Snippet: S1P-induced cytoplasmic membrane localization of Endoglin dependent on ERK activation in endothelial cells. (A) HPMVECs were treated with BSA or 200 nM S1P for 30 min and then were fixed prior to immunofluorescent staining of Endoglin. Nuclei were stained with DAPI. Red arrows indicate the enrichment of Endoglin on the plasma membrane. Magnification of BSA or S1P group, top panel 100×, lower panel 400×; Scale bars, 200 μm. (B) HPMVECs incubated with DMSO or 1 μM SCH772984 for 30 min were treated with S1P after words. The cells were processed as described in panel A and analyzed with fluorescent microscopy. Magnification, 400×; Scale bars, 200 μm. S1P, sphingosine 1-phosphate; ERK, extracellular regulated protein kinases; HPMVEC, human pulmonary microvascular endothelial cell; BSA, bovine serum albumin; DAPI, 4, 6'-diamidino-2-phenylindole; DMSO, dimethyl sulfoxide; SCH772984, a specific ERK inhibitor.

Article Snippet: HPMVEC (human pulmonary microvascular endothelial cell) was purchased from Lonza (Portsmouth, USA).

Techniques: Membrane, Activation Assay, Staining, Clinical Proteomics, Incubation, Microscopy