Journal: The American Journal of Tropical Medicine and Hygiene

Article Title: Dengue Virus Infection of Primary Human Smooth Muscle Cells

doi: 10.4269/ajtmh.18-0175

Figure Lengend Snippet: Human umbilical artery smooth muscle cells (HUASMC) are permissive to dengue virus (DENV) infection. Virion production and release was quantified by plaque assays of culture supernatants at different time points post-infection (p.i.) of HUASMC, human umbilical vein endothelial cells (HUVEC), and LLC-MK2 (macaque kidney cells) infected with each of the four DENV serotypes (multiplicity of infection:1). (A) Infection kinetics of HUASMC cells (24–72 hours p.i.) with DENV 1–4 measured by plaque assays (plaque-forming units [PFU]/mL). (B) DENV 1–4 titers in culture supernatants of HUASMC, HUVEC, and LLC-MK2 cells at 72 hours p.i. Data are expressed as the mean ± standard deviation of three independent experiments. ***P < 0.001 compared with its HUVEC and LLC-MK2 cells counterparts. (D) l. = assay detection limit.

Article Snippet: Human umbilical artery smooth muscle cells and HUVEC were purchased and maintained in smooth muscle cell growth medium and endothelial cell growth medium, respectively, according to the manufacturer’s instructions (Cell Applications, San Diego, CA).

Techniques: Virus, Infection, Standard Deviation

Journal: The American Journal of Tropical Medicine and Hygiene

Article Title: Dengue Virus Infection of Primary Human Smooth Muscle Cells

doi: 10.4269/ajtmh.18-0175

Figure Lengend Snippet: Human umbilical artery smooth muscle cells (HUASMC) release dengue virus (DENV) genomes on infection. Dengue virus genomic RNA was quantified by real-time qRT-PCR from cell culture supernatants of HUASMC, human umbilical vein endothelial cells (HUVEC), and LLC-MK2 (macaque kidney cells) infected with DENV (multiplicity of infection:1). (A) Dengue virus 1–4 infection kinetics (24–72 hours post-infection [p.i.]) of HUASMC cells measured by real-time genomic qRT-PCR (copies/mL). (B) Genome copies present in culture supernatants of HUASMC, HUVEC, and LLC-MK2 cells at 72 hours p.i. with the four DENV serotypes. (C) Calculated genome-to-plaque-forming unit (PFU) ratios of HUASMC, HUVEC, and LLC-MK2 cells supernatants at 72 hours p.i. with each DENV serotype. Data are expressed as the mean ± standard deviation of three independent experiments. *P < 0.05, **P < 0.005, and ***P < 0.001 calculated to its HUASMC counterpart.

Article Snippet: Human umbilical artery smooth muscle cells and HUVEC were purchased and maintained in smooth muscle cell growth medium and endothelial cell growth medium, respectively, according to the manufacturer’s instructions (Cell Applications, San Diego, CA).

Techniques: Virus, Infection, Quantitative RT-PCR, Cell Culture, Standard Deviation

Journal: The American Journal of Tropical Medicine and Hygiene

Article Title: Dengue Virus Infection of Primary Human Smooth Muscle Cells

doi: 10.4269/ajtmh.18-0175

Figure Lengend Snippet: Dengue virus (DENV) antigens are detected in human umbilical artery smooth muscle cells (HUASMC). Epifluorescence images of immunostained cells with an anti-DENV 1-4 envelope protein-specific monoclonal antibody (green), and a cytoplasmic (red) and nuclei (blue) counterstains. The images were captured at 72 hours post-infection (p.i.) with each DENV serotype at a multiplicity of infection (MOI) of 1. (A) Representative image of mock-infected HUASMC cells at ×100 magnification (scale bar = 50 μm). (B and C) Representative images of DENV-infected HUASMC (arrows) at ×400 (scale bar = 20 μm) and ×100 (scale bar = 50 μm) magnification, respectively. (D) Image analysis for quantifying infected cells (green outline) vs total cell nuclei (white outlines) with the software CellProfiler 2.0. (E) Percentages of infected cells in HUASMC, human umbilical vein endothelial cells (HUVEC), and LLC-MK2 (macaque kidney cells) cell lines. (F) Comparison of DENV labeling by immunostaining with an anti-DENV 1-4 envelope protein-specific monoclonal antibody (green) and an anti-NS3 polyclonal antibody (red) in HUASMC cells at 72 hours p.i. with DENV-2 and DENV-3 at a MOI of 1. Magnification of ×400 (scale bar = 20 μm). Data are expressed as mean ± standard deviation of three independent experiments. **P < 0.005, ***P < 0.001 calculated to its HUASMC cells counterpart. This figure appears in color at www.ajtmh.org.

Article Snippet: Human umbilical artery smooth muscle cells and HUVEC were purchased and maintained in smooth muscle cell growth medium and endothelial cell growth medium, respectively, according to the manufacturer’s instructions (Cell Applications, San Diego, CA).

Techniques: Virus, Infection, Software, Comparison, Labeling, Immunostaining, Standard Deviation

Journal: British Journal of Pharmacology

Article Title: Serelaxin-mediated signal transduction in human vascular cells: bell-shaped concentration–response curves reflect differential coupling to G proteins

doi: 10.1111/bph.12964

Figure Lengend Snippet: The expression of RXFP1 and RXFP2 receptor mRNA and RXFP1 receptor protein in human primary umbilical vascular cells and human primary cardiac fibroblasts. qPCR (A) was utilized to show expression levels of RXFP1 and RXFP2 receptor mRNA in HUAECs, HUVECs, HUASMCs, HUVSMCs and HCFs relative to β-actin (n = 2). RXFP2 receptor mRNA was only measureable in the positive control. Cell surface RXFP1 receptor protein expression was determined by radioligand binding (B) utilizing [125I]-serelaxin and showed specific serelaxin binding in HEK-RXFP1 cells (n = 6), HUASMCs (n = 4), HUVECs (n = 4), HUVSMCs (n = 3) and HCFs (n = 3), but not in HUAECs (n = 2).

Article Snippet: Cell culture Primary cultures of human umbilical artery endothelial cells (HUAECs), HUVECs, human umbilical artery smooth muscle cells (HUASMCs), human umbilical vein smooth muscle cells (HUVSMCs) and fetal human cardiac fibroblasts (HCFs: pooled from fetal atria and ventricles) were obtained from ScienCell Research Laboratories (San Diego, CA, USA).

Techniques: Expressing, Positive Control, Binding Assay

Journal: British Journal of Pharmacology

Article Title: Serelaxin-mediated signal transduction in human vascular cells: bell-shaped concentration–response curves reflect differential coupling to G proteins

doi: 10.1111/bph.12964

Figure Lengend Snippet: The effect of serelaxin on cAMP accumulation in human primary umbilical vascular cells and cardiac fibroblasts. Serelaxin treatment (30 min) increased cAMP accumulation in (A) HUVECs (n = 6), (B) HUVSMCs (n = 6) and (C) HUASMCs (n = 4), but not in (D) HCFs (n = 3). The serelaxin CRC was bell-shaped for HUVECs and HUVSMCs but sigmoidal for HUASMCs. For each cell type, the effect of PTX (50 ng·mL−1, 18 h) and wortmannin (100 nM, 30 min) pretreatment was determined after exposure to serelaxin (30 nM) for 30 min to determine the role of Gαi and PI3K. Statistical significance was assessed using a one-way anova with a Dunnett's post hoc test compared with serelaxin alone: *P < 0.05 and **P < 0.01.

Article Snippet: Cell culture Primary cultures of human umbilical artery endothelial cells (HUAECs), HUVECs, human umbilical artery smooth muscle cells (HUASMCs), human umbilical vein smooth muscle cells (HUVSMCs) and fetal human cardiac fibroblasts (HCFs: pooled from fetal atria and ventricles) were obtained from ScienCell Research Laboratories (San Diego, CA, USA).

Techniques:

Journal: British Journal of Pharmacology

Article Title: Serelaxin-mediated signal transduction in human vascular cells: bell-shaped concentration–response curves reflect differential coupling to G proteins

doi: 10.1111/bph.12964

Figure Lengend Snippet: The effect of serelaxin on cGMP accumulation in human primary umbilical vascular cells and cardiac fibroblasts. Serelaxin treatment (30 min) increased cGMP accumulation in (A) HUVECs (n = 7), (B) HUVSMCs (n = 5), (C) HUASMCs (n = 6) and (D) HCFs (n = 5). The serelaxin CRC was bell-shaped for HUVECs, HUVSMCs and HCFs but sigmoidal for HUASMCs. PTX (50 ng·mL−1, 18 h) and wortmannin (100 nM, 30 min) pretreatment significantly inhibited serelaxin (30 nM)-mediated cGMP accumulation in each cell type. Statistical significance was assessed using a one-way anova with a Dunnett's post hoc test compared with serelaxin alone: *P < 0.05 and **P < 0.01.

Article Snippet: Cell culture Primary cultures of human umbilical artery endothelial cells (HUAECs), HUVECs, human umbilical artery smooth muscle cells (HUASMCs), human umbilical vein smooth muscle cells (HUVSMCs) and fetal human cardiac fibroblasts (HCFs: pooled from fetal atria and ventricles) were obtained from ScienCell Research Laboratories (San Diego, CA, USA).

Techniques:

Journal: British Journal of Pharmacology

Article Title: Serelaxin-mediated signal transduction in human vascular cells: bell-shaped concentration–response curves reflect differential coupling to G proteins

doi: 10.1111/bph.12964

Figure Lengend Snippet: Changes in the expression of nNOS, VEGF and ETB receptors in human primary umbilical vascular cells and cardiac fibroblasts after serelaxin (1.68 nM) exposure for 24 and 48 h. In HUVECs (A), serelaxin treatment increased the expression of nNOS (n = 5), ETB receptors (n = 6) and VEGF (n = 7). In HUVSMCs (B), serelaxin treatment increased the expression of nNOS (n = 5) and ETB (n = 5) but not VEGF (n = 4); however, in HUASMC (C), serelaxin treatment increased the expression of nNOS (n = 6), ETB receptors (n = 7) and VEGF (n = 5). In HCFs (D), similar to HUVECs and HUASMCs, serelaxin treatment increased the expression of nNOS (n = 5), ETB receptors (n = 5) and VEGF (n = 5). A representative blot of each protein and β-actin, a loading control, is shown with the densitometry in each figure. Statistical significance was assessed using a one-way anova with a Dunnett's post hoc test compared with vehicle alone: *P < 0.05 and **P < 0.01.

Article Snippet: Cell culture Primary cultures of human umbilical artery endothelial cells (HUAECs), HUVECs, human umbilical artery smooth muscle cells (HUASMCs), human umbilical vein smooth muscle cells (HUVSMCs) and fetal human cardiac fibroblasts (HCFs: pooled from fetal atria and ventricles) were obtained from ScienCell Research Laboratories (San Diego, CA, USA).

Techniques: Expressing, Control

Journal: British Journal of Pharmacology

Article Title: Serelaxin-mediated signal transduction in human vascular cells: bell-shaped concentration–response curves reflect differential coupling to G proteins

doi: 10.1111/bph.12964

Figure Lengend Snippet: Changes in the activity of MMPs in human primary umbilical vascular cells and cardiac fibroblasts using zymography to assess changes in activity of MMP2 and MMP9 after long-term serelaxin exposure (1.68 and 30 nM) for 48 h. Serelaxin treatment increased the activity of MMP2 in (A) HUVECs (n = 5), (B) HUVSMCs (n = 6), (C) HUASMCs (n = 7) and (D) HCFs (n = 5). Serelaxin also significantly increased the activity of MMP9 but only in (B) HUVSMCs (n = 5) and (D) HCFs (n = 5) and not in (A) HUVECs (n = 5) and (C) HUASMCs (n = 5). Furthermore, and consistent with the lack of RXFP1 receptor expression (Figure 1), serelaxin had no effect on MMP activity in HUAECs (n = 2). A representative scan of each zymography is shown along with the densitometry in each figure. Statistical significance was assessed using a one-way anova with a Dunnett's post hoc test compared with control alone: *P < 0.05 and **P < 0.01.

Article Snippet: Cell culture Primary cultures of human umbilical artery endothelial cells (HUAECs), HUVECs, human umbilical artery smooth muscle cells (HUASMCs), human umbilical vein smooth muscle cells (HUVSMCs) and fetal human cardiac fibroblasts (HCFs: pooled from fetal atria and ventricles) were obtained from ScienCell Research Laboratories (San Diego, CA, USA).

Techniques: Activity Assay, Zymography, Expressing, Control

Journal: British Journal of Pharmacology

Article Title: Serelaxin-mediated signal transduction in human vascular cells: bell-shaped concentration–response curves reflect differential coupling to G proteins

doi: 10.1111/bph.12964

Figure Lengend Snippet: Signal transduction mechanisms employed by serelaxin in human umbilical vascular cells and HCFs and their potential physiological effects. Short-term (<1 h) serelaxin stimulation produces cAMP/cGMP accumulation in vascular cells and pERK1/2 in all cells that is differentially regulated by Gαs, Gαi, GαOB and PI3K, and these pathways are likely to be involved in the vasodilator and anti-apoptotic effects of serelaxin respectively. In HUASMCs, the serelaxin-mediated cAMP or VEGF response did not involve GαOB and PI3K, whereas in HCFs, the RXFP1 receptor was not coupled to cAMP production. Longer term (24–48 h) serelaxin treatment increased VEGF expression involving both cAMP-dependent and cAMP-independent mechanisms, and these pathways are likely to be involved in angiogenesis. In addition, serelaxin treatment increased the activity of MMP2 and MMP9 to mediate its remodelling actions, but these enzymes are also secreted to convert big ET to ET1-32 that activates ETB receptors to further enhance vasodilatation (Conrad, 2010). Solid lines indicate mechanisms identified in the current study whereas dashed lines indicate previously established mechanisms.

Article Snippet: Cell culture Primary cultures of human umbilical artery endothelial cells (HUAECs), HUVECs, human umbilical artery smooth muscle cells (HUASMCs), human umbilical vein smooth muscle cells (HUVSMCs) and fetal human cardiac fibroblasts (HCFs: pooled from fetal atria and ventricles) were obtained from ScienCell Research Laboratories (San Diego, CA, USA).

Techniques: Transduction, Expressing, Activity Assay

Journal:

Article Title: Development of Composite Porous Scaffolds Based on Collagen and Biodegradable Poly(ester urethane)urea

doi:

Figure Lengend Snippet: Growth curves for scaffolds made from PEUU (C0/ 1080) and PEUU with 10% collagen (C10/1080) seeded with umbilical artery smooth muscle cells and cultured for up to 28 days. The relative cell number is based upon MTT absorption and reflects mitochondrial activity.

Article Snippet: Scaffolds were cut and sterilized by immersion in 70% ethanol for 6 h, followed by rinsing with PBS and exposure to the ultraviolet light source in a laminar flow hood (Class II A/B3 Biological Safety Cabinet) for 2 h. Scaffolds were placed in a centrifuge tube with human umbilical artery smooth muscle cells (UASMCs, Bio Whittaker) at a density of 1 × 10 6 cells/ml in medium (SmGM-2 with 10% fetal bovine serum, BioWhittaker).

Techniques: Cell Culture, Activity Assay

Journal:

Article Title: Development of Composite Porous Scaffolds Based on Collagen and Biodegradable Poly(ester urethane)urea

doi:

Figure Lengend Snippet: Electron micrographs of PEUU and PEUU/collagen (90:10) scaffold surfaces after 28 days of culture with umbilical artery smooth muscle cells.

Article Snippet: Scaffolds were cut and sterilized by immersion in 70% ethanol for 6 h, followed by rinsing with PBS and exposure to the ultraviolet light source in a laminar flow hood (Class II A/B3 Biological Safety Cabinet) for 2 h. Scaffolds were placed in a centrifuge tube with human umbilical artery smooth muscle cells (UASMCs, Bio Whittaker) at a density of 1 × 10 6 cells/ml in medium (SmGM-2 with 10% fetal bovine serum, BioWhittaker).

Techniques:

Journal: British Journal of Pharmacology

Article Title: Enhanced serelaxin signalling in co‐cultures of human primary endothelial and smooth muscle cells

doi: 10.1111/bph.13371

Figure Lengend Snippet: cGMP accumulation in co‐cultures of human primary vascular smooth muscle cells following addition of serelaxin to endothelium. HUAEC , HUVEC or HCAEC were co‐cultured with (A) HUASMC or (B) HUVSMC (all n = 5), and the ECs were treated with serelaxin for 30 min. Serelaxin addition to HUAEC did not cause cGMP accumulation in HUAEC (▲) (C) HUASMC (□) or (D) HUVSMC (◯) co‐cultured with HUAEC, whereas direct stimulation of either (C) HUASMC (n = 5) or (D) HUVSMC with serelaxin caused a concentration‐dependent increase in cGMP accumulation (dashed lines). In contrast, serelaxin addition to HUVEC concentration‐dependently increased cGMP accumulation not only in HUVEC (■) but also in (E) HUASMC (□) or (F) HUVSMC (◯) co‐cultured with HUVEC with the responses in smooth muscle cells being greater or in the case of HUVSMC much greater than cGMP responses to direct stimulation of (E) HUASMC or (F) HUVSMC (dashed lines). A similar pattern of cGMP accumulation was observed with (G, H) HCAEC (●) and (G) HUASMC (□) or (H) HUVSMC (◯) co‐cultured with HCAEC.

Article Snippet: Primary cultures of human umbilical artery endothelial cells (HUAEC), HUVEC, human coronary artery endothelial cells (HCAEC), human umbilical artery smooth muscle cells (HUASMC) and human umbilical vein smooth muscle cells (HUVSMC) were obtained from ScienCell Research Laboratories (San Diego, CA, USA ).

Techniques: Cell Culture, Concentration Assay

Journal: British Journal of Pharmacology

Article Title: Enhanced serelaxin signalling in co‐cultures of human primary endothelial and smooth muscle cells

doi: 10.1111/bph.13371

Figure Lengend Snippet: Serelaxin‐mediated cGMP accumulation in monocultures of human primary vascular cells (all n = 5). Serelaxin (30 nM, 30 min) increased cGMP accumulation in (A) HUVEC, (B) HCAEC, (C) HUASMC and (D) HUVSMC. Pre‐incubation with l‐NOARG (30 μM, 30 min) or ODQ (1 μM, 30 min) almost abolished serelaxin‐mediated (30 nM, 30 min) cGMP accumulation in all cell types. Pre‐treatment with indomethacin (30 μM, 30 min) significantly inhibited serelaxin‐mediated (30 nM, 30 min) cGMP accumulation in (B) HCAEC but had no effect in (A) HUVEC, (C) HUASMC or (D) HUVSMC. *P < 0.05, **P < 0.02, ***P < 0.005; significantly different from serelaxin alone; one‐way anova with Dunnett's post hoc test.

Article Snippet: Primary cultures of human umbilical artery endothelial cells (HUAEC), HUVEC, human coronary artery endothelial cells (HCAEC), human umbilical artery smooth muscle cells (HUASMC) and human umbilical vein smooth muscle cells (HUVSMC) were obtained from ScienCell Research Laboratories (San Diego, CA, USA ).

Techniques: Incubation

Journal: British Journal of Pharmacology

Article Title: Enhanced serelaxin signalling in co‐cultures of human primary endothelial and smooth muscle cells

doi: 10.1111/bph.13371

Figure Lengend Snippet: Serelaxin‐mediated cGMP accumulation in human primary vascular smooth muscle cells co‐cultured with HUVEC or (A) HCAEC (all n = 6 except where otherwise indicated). Stimulation of HUVEC or HCAEC with serelaxin (30 nM, 30 min) increased cGMP accumulation not only in (B) HUVEC and (C) HCAEC but also in co‐cultures of (D, F) HUASMC or (E, G) HUVSMC. Pre‐incubation of HUVEC or HCAEC with l‐NOARG (30 μM, 30 min) before addition of serelaxin (30 nM, 30 min) significantly inhibited cGMP accumulation not only in HUVEC and (C) HCAEC but also in (D, F) HUASMC and (E, G) HUVSMC. Pre‐incubation of HUVEC with indomethacin (30 μM, 30 min) did not affect serelaxin‐mediated (30 nM, 30 min) cGMP accumulation in (B) HUVEC or in co‐incubated (D) HUASMC or (E) HUVSMC (n = 5). Pre‐incubation of HCAEC with indomethacin (30 μM, 30 min) had no significant effect on serelaxin‐mediated (30 nM, 30 min) cGMP accumulation in (C) HCAEC but produced marked and significant reductions in cGMP accumulation in co‐incubated (F) HUASMC or (G) HUVSMC (n = 5). Pre‐treatment of HUASMC or HUVSMC with ODQ (1 μM, 30 min) had no significant effect on serelaxin‐mediated (30 nM, 30 min) cGMP accumulation in (I) HUVEC or (J) HCAEC but reduced or abolished cGMP accumulation in (K, M) HUASMC or (L, N) HUVSMC (n = 5). *P < 0.05, **P < 0.02, ***P < 0.005 significantly different from serelaxin alone; one‐way anova with Dunnett's post hoc test.

Article Snippet: Primary cultures of human umbilical artery endothelial cells (HUAEC), HUVEC, human coronary artery endothelial cells (HCAEC), human umbilical artery smooth muscle cells (HUASMC) and human umbilical vein smooth muscle cells (HUVSMC) were obtained from ScienCell Research Laboratories (San Diego, CA, USA ).

Techniques: Cell Culture, Incubation, Produced

Journal: British Journal of Pharmacology

Article Title: Enhanced serelaxin signalling in co‐cultures of human primary endothelial and smooth muscle cells

doi: 10.1111/bph.13371

Figure Lengend Snippet: cAMP accumulation in co‐cultures of human primary vascular smooth muscle cells following addition of serelaxin to endothelium (all n = 5). HUAEC, HUVEC or HCAEC were co‐cultured with (A) HUASMC or (B) HUVSMC, and the endothelial cells were treated with serelaxin for 30 min. Serelaxin added to HUAEC did not cause cAMP accumulation either in (C, D) HUAEC (▲), (C) HUASMC (□) or (D) HUVSMC (◯), whereas direct stimulation of (C) HUASMC or (D) HUVSMC with serelaxin caused a concentration‐dependent increase in cAMP accumulation (dashed lines). Although direct addition of serelaxin to HUVEC concentration‐dependently increased cAMP accumulation in (E, F) HUVEC (■), there was no significant effect on cAMP accumulation in (E) HUASMC (□) or (F) HUVSMC (◯). Direct addition of serelaxin to (E) HUASMC or (F) HUVSMC stimulated cAMP accumulation (dashed lines). Serelaxin concentration‐dependently increased cAMP accumulation in (G, H) HCAEC (●) but also caused a robust concentration‐dependent increase in cAMP accumulation in both (G) HUASMC (□) and (H) HUVSMC (◯).

Article Snippet: Primary cultures of human umbilical artery endothelial cells (HUAEC), HUVEC, human coronary artery endothelial cells (HCAEC), human umbilical artery smooth muscle cells (HUASMC) and human umbilical vein smooth muscle cells (HUVSMC) were obtained from ScienCell Research Laboratories (San Diego, CA, USA ).

Techniques: Cell Culture, Concentration Assay

Journal: British Journal of Pharmacology

Article Title: Enhanced serelaxin signalling in co‐cultures of human primary endothelial and smooth muscle cells

doi: 10.1111/bph.13371

Figure Lengend Snippet: Serelaxin‐mediated cAMP accumulation in monocultures of human primary vascular cells (all n = 5). Serelaxin (30 nM, 30 min) increased cAMP accumulation in (A) HUVEC, (B) HCAEC, (C) HUASMC and (D) HUVSMC that was not significantly altered by pre‐incubation with l‐NOARG (30 μM, 30 min) or ODQ (1 μM, 30 min). Pre‐treatment with indomethacin (30 μM, 30 min) significantly inhibited serelaxin‐mediated (30 nM, 30 min) cAMP accumulation in (B) HCAEC but not in (A) HUVEC, (C) HUASMC or (D) HUVSMC. *P < 0.05; significantly different from serelaxin alone; one‐way anova with Dunnett's post hoc test.

Article Snippet: Primary cultures of human umbilical artery endothelial cells (HUAEC), HUVEC, human coronary artery endothelial cells (HCAEC), human umbilical artery smooth muscle cells (HUASMC) and human umbilical vein smooth muscle cells (HUVSMC) were obtained from ScienCell Research Laboratories (San Diego, CA, USA ).

Techniques: Incubation

Journal: British Journal of Pharmacology

Article Title: Enhanced serelaxin signalling in co‐cultures of human primary endothelial and smooth muscle cells

doi: 10.1111/bph.13371

Figure Lengend Snippet: Serelaxin‐mediated cAMP accumulation in human primary vascular smooth muscle cells co‐cultured with HCAEC (A, E; all n = 5). Stimulation of HCAEC with serelaxin (30 nM, 30 min) increased cAMP accumulation not only in (B) HCAEC but also in co‐cultures of (C) HUASMC or (D) HUVSMC. Pre‐incubation of HCAEC with l‐NOARG (30 μM, 30 min) before addition of serelaxin (30 nM, 30 min) had no significant effect on cAMP accumulation in (B) HCAEC, (C) HUASMC or (D) HUVSMC. However, pre‐incubation of HCAEC with indomethacin (30 μM, 30 min) significantly inhibited serelaxin‐mediated (30 nM, 30 min) cAMP accumulation in (B) HCAEC and abolished cAMP accumulation in (C) HUASMC or (D) HUVSMC. Pre‐treatment of HUASMC or HUVSMC with ODQ (1 μM, 30 min) had no significant effect on serelaxin‐mediated (30 nM, 30 min) cAMP accumulation in (F) HCAEC, (G) HUASMC or (H) HUVSMC. *P < 0.05; significantly different from serelaxin alone; one‐way anova with Dunnett's post hoc test.

Article Snippet: Primary cultures of human umbilical artery endothelial cells (HUAEC), HUVEC, human coronary artery endothelial cells (HCAEC), human umbilical artery smooth muscle cells (HUASMC) and human umbilical vein smooth muscle cells (HUVSMC) were obtained from ScienCell Research Laboratories (San Diego, CA, USA ).

Techniques: Cell Culture, Incubation

Journal: British Journal of Pharmacology

Article Title: Enhanced serelaxin signalling in co‐cultures of human primary endothelial and smooth muscle cells

doi: 10.1111/bph.13371

Figure Lengend Snippet: Signal transduction mechanisms activated by serelaxin in co‐cultures of human primary vascular cells. Activation of RXFP1 by serelaxin in HUVEC and HCAEC stimulates NO production and activates sGC and AC to produce cGMP and cAMP respectively. Endothelial NO also diffuses from the endothelial cells across the ThinCert membranes and activates sGC in both the arterial and venous smooth muscle cells. Additionally in HCAEC (blue lines) but not HUVEC, serelaxin stimulates prostanoid production that produces cAMP accumulation in both arterial and smooth muscle cells.

Article Snippet: Primary cultures of human umbilical artery endothelial cells (HUAEC), HUVEC, human coronary artery endothelial cells (HCAEC), human umbilical artery smooth muscle cells (HUASMC) and human umbilical vein smooth muscle cells (HUVSMC) were obtained from ScienCell Research Laboratories (San Diego, CA, USA ).

Techniques: Transduction, Activation Assay