Journal: International Journal of Environmental Research and Public Health

Article Title: A Muscarinic Antagonist Reduces Airway Inflammation and Bronchoconstriction Induced by Ambient Particulate Matter in a Mouse Model of Asthma

doi: 10.3390/ijerph15061189

Figure Lengend Snippet: Experimental protocol of the mouse model of ovalbumin (OVA)-induced asthma used in the present study. For details, please see the text. FORM, formoterol fumarate; FP, fluticasone propionate; NS, normal saline; PM, particulate matter; TIO, tiotropium bromide.

Article Snippet: To investigate the effect of drugs on airway inflammation and respiratory function, mice were treated with fluticasone propionate (Toronto Research Chemicals Inc., North York, ON, Canada), formoterol fumarate (Toronto Research Chemicals Inc.), or tiotropium bromide (Tokyo Chemical Industry Co., Ltd., Tokyo, Japan) on days 21 to 26.

Techniques: Saline

Journal: International Journal of Environmental Research and Public Health

Article Title: A Muscarinic Antagonist Reduces Airway Inflammation and Bronchoconstriction Induced by Ambient Particulate Matter in a Mouse Model of Asthma

doi: 10.3390/ijerph15061189

Figure Lengend Snippet: Effects of fluticasone propionate, formoterol fumarate, and tiotropium bromide on airway resistance. Airway resistance was evaluated by specific airway resistance (sRaw) values on day 27. Formoterol fumarate and tiotropium bromide decreased the sRaw values compared with fluticasone propionate. Data for each group are expressed as the mean ± standard deviation, with eight mice per group. * p < 0.05.

Article Snippet: To investigate the effect of drugs on airway inflammation and respiratory function, mice were treated with fluticasone propionate (Toronto Research Chemicals Inc., North York, ON, Canada), formoterol fumarate (Toronto Research Chemicals Inc.), or tiotropium bromide (Tokyo Chemical Industry Co., Ltd., Tokyo, Japan) on days 21 to 26.

Techniques: Standard Deviation

Journal: International Journal of Environmental Research and Public Health

Article Title: A Muscarinic Antagonist Reduces Airway Inflammation and Bronchoconstriction Induced by Ambient Particulate Matter in a Mouse Model of Asthma

doi: 10.3390/ijerph15061189

Figure Lengend Snippet: The levels of reactive oxygen metabolites after the administration of fluticasone propionate, formoterol fumarate, and tiotropium bromide. The levels of reactive oxygen metabolites (dROMs) in serum samples obtained on day 27. Fluticasone propionate and tiotropium bromide had no effect on dROM levels compared with the control group, but formoterol fumarate increased dROM levels. Data for each group are expressed as the mean ± standard deviation, with eight mice per group. * p < 0.05.

Article Snippet: To investigate the effect of drugs on airway inflammation and respiratory function, mice were treated with fluticasone propionate (Toronto Research Chemicals Inc., North York, ON, Canada), formoterol fumarate (Toronto Research Chemicals Inc.), or tiotropium bromide (Tokyo Chemical Industry Co., Ltd., Tokyo, Japan) on days 21 to 26.

Techniques: Control, Standard Deviation

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Tobacco Smoke and Inhaled Drugs Alter Expression and Activity of Multidrug Resistance-Associated Protein-1 (MRP1) in Human Distal Lung Epithelial Cells in vitro

doi: 10.3389/fbioe.2020.01030

Figure Lengend Snippet: Influence of Inhaled drugs on multidrug resistance-associated protein-1 (MRP1) activity in NCI-H441 cells.

Article Snippet: As described above, cell monolayers were loaded with CFDA solution alone or containing either budesonide (5 or 10 μM), beclomethasone dipropionate (50 μM, Sigma-Aldrich), salbutamol sulfate (100 μM), salbutamol base (100 μM, Sigma-Aldrich), R-salbutamol HCl (100 μM, Sunovion Pharmaceuticals, Marlborough, MA, United States), S-salbutamol HCl, (100 μM, Sunovion Pharmaceuticals), terbutaline (100 μM, Sunovion Pharmaceuticals), formoterol fumarate (100 μM, Santa Cruz), L-sulforaphane (10 μM, Cayman Chemical, Ann Arbor, MI, United States), 5 or 10% CSE or the solvent [i.e., dimethyl sulfoxide (DMSO)] when appropriate.

Techniques: Activity Assay

Journal: Drug testing and analysis

Article Title: Enantioselective disposition of (R,R)-formoterol, (S,S)-formoterol and their respective glucuronides in urine following single inhaled dosing and application to doping control.

doi: 10.1002/dta.2587

Figure Lengend Snippet: Figure 2. Urine levels (mean±SEM) of formoterol enantiomers (A) and formoterol enantiomer glucuronides (B) following a 12 μg inhaled dose of rac-formoterol demonstrating enantioselective pharmacokinetics.

Article Snippet: In brief, calibration samples were prepared consisting of concentrations of 0.1, 0.5, 2.0, 10, 40 ng/mL unlabeled formoterol in drug free human urine from rac-formoterol fumarate dihydrate (Carbosynth, Compton, UK).

Techniques: Drug discovery

Journal: Drug testing and analysis

Article Title: Enantioselective disposition of (R,R)-formoterol, (S,S)-formoterol and their respective glucuronides in urine following single inhaled dosing and application to doping control.

doi: 10.1002/dta.2587

Figure Lengend Snippet: Figure 3. Extent of conjugation (free formoterol as a proportion of total) for each enantiomer.

Article Snippet: In brief, calibration samples were prepared consisting of concentrations of 0.1, 0.5, 2.0, 10, 40 ng/mL unlabeled formoterol in drug free human urine from rac-formoterol fumarate dihydrate (Carbosynth, Compton, UK).

Techniques: Conjugation Assay

Journal: Drug testing and analysis

Article Title: Enantioselective disposition of (R,R)-formoterol, (S,S)-formoterol and their respective glucuronides in urine following single inhaled dosing and application to doping control.

doi: 10.1002/dta.2587

Figure Lengend Snippet: Figure 4. Enantioselectivity of formoterol (free) and formoterol glucuronide (conjugated) in urine following a 12 μg inhaled dose

Article Snippet: In brief, calibration samples were prepared consisting of concentrations of 0.1, 0.5, 2.0, 10, 40 ng/mL unlabeled formoterol in drug free human urine from rac-formoterol fumarate dihydrate (Carbosynth, Compton, UK).

Techniques:

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Medium-Dose Formoterol Attenuated Abdominal Aortic Aneurysm Induced by EPO via β2AR/cAMP/SIRT1 Pathway.

doi: 10.1002/advs.202306232

Figure Lengend Snippet: Figure 1. Analysis of differential expression genes in GSE174556 dataset. A) Flow chart shows our bioinformatic and experimental strategy. GSE174556 dataset was composed of the RNA-seq data of entire aortas isolated from ApoE−/−mice receiving EPO (n = 3) and vehicle (n = 3) treatment. We conducted differential expression genes (DEGs) and GO&KEGG enrichment analyses on the dataset. AAA-centered dataset provided by Phenopedia intersected with GSE174556 dataset. Formoterol was chosen by CMap as the compound to confront EPO-induced-AAA in later experiments. We testified the effect of formoterol by injection of formoterol to ApoE−/−mice in EPO-induced AAA model. B) Heatmap of all DEGs in GSE174556 dataset resulting from a two-way hierarchical clustering. C) Volcano map of DEGs. The blue dots represent the downregulated DEGs and the red ones represent the upregulated DEGs. D) The number of significantly upregulated and downregulated genes. E) Gene ontology (GO) enrichment for DEGs in GSE174556 dataset. F) Kyoto Encyclopedia of genes and genomes (KEGG) enrichment for DEGs in GSE174556 dataset.

Article Snippet: In the first part of the in vitro experiments, to examine the dose-response relation between formoterol treatment and VSMC senescence, VSMC were divided into 6 groups, which were treated with vehicle (PBS), 5IU mL−1 EPO, 5IU mL−1 EPO+0.01 nmol mL−1 formoterol (S2020, Selleck, USA), 5IU mL−1 EPO+0.1 nmol mL−1 formoterol, 5IU mL−1 EPO+1 nmol mL−1 formoterol and 5IU mL−1 EPO+10 nmol mL−1 formoterol, respectively.

Techniques: Expressing, RNA Sequencing, Isolation, Injection

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Medium-Dose Formoterol Attenuated Abdominal Aortic Aneurysm Induced by EPO via β2AR/cAMP/SIRT1 Pathway.

doi: 10.1002/advs.202306232

Figure Lengend Snippet: Figure 2. Formoterol (FORM) suppressed the formation of EPO-induced AAA in ApoE−/−mice. A) Venn diagram showing the intersection of DEGs and AAA-centered gene set provided by Phenopedia B) Venn diagram showing that formoterol was the only option (number “1” in Figure 2B) in the intersection among Connectivity map results based on all DEGs, upregulated DEGs and downregulated DEGs. C) Representative photographs of the abdominal aortic specimens in ApoE−/−mice receiving vehicle, EPO, EPO+low-dose formoterol, EPO+medium-dose formoterol, EPO+high-dose for- moterol treatment, respectively. D)The incidence of AAA in each group (n = 20 per group). E) Abdominal aorta diameters in each group (n = 8 per group). F) The survival rate in each group (n = 20 per group). G) Representative H&E staining and Verhoeffstaining of abdominal aortic section in ApoE−/−mice receiving vehicle, EPO, EPO+low-dose formoterol, EPO+medium-dose formoterol, EPO+high-dose formoterol treatment, respectively. Low (×4, scale bars = 100 μm), and medium (×20, scale bars = 25 μm), magnifications are shown. H) Representative western blot analysis of collagen I (COL I) and collagen III (COL III) expression in abdominal aortas of 5 groups of mice. I-J) Quantitative analysis of COL I and COL III expression in abdominal aortas of 5 groups of mice (n = 6 per group). Fisher’s exact test was used for (D). The Kruskal-Wallis test followed by Nemenyi post hoc test was used for (E). Log-rank (Mantel-Cox) test was used for (F). The other data were analyzed via One-way ANOVA followed by the Tukey test for post hoc comparison. *p < 0.05, **p <0.01, ***p <0.001, mean ± SEM.

Article Snippet: In the first part of the in vitro experiments, to examine the dose-response relation between formoterol treatment and VSMC senescence, VSMC were divided into 6 groups, which were treated with vehicle (PBS), 5IU mL−1 EPO, 5IU mL−1 EPO+0.01 nmol mL−1 formoterol (S2020, Selleck, USA), 5IU mL−1 EPO+0.1 nmol mL−1 formoterol, 5IU mL−1 EPO+1 nmol mL−1 formoterol and 5IU mL−1 EPO+10 nmol mL−1 formoterol, respectively.

Techniques: Staining, Western Blot, Expressing, Comparison

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Medium-Dose Formoterol Attenuated Abdominal Aortic Aneurysm Induced by EPO via β2AR/cAMP/SIRT1 Pathway.

doi: 10.1002/advs.202306232

Figure Lengend Snippet: Figure 3. Effect of formoterol (FORM) on expression of matrix metalloproteinases (MMPs) and proinflammatory cytokines in EPO-induced AAA. A) Representative immunohistochemical staining of the abdominal aortic sections for MMP2, MMP9, VCAM, MCP1, and IL-6 in 5 groups of mice (scale bars = 25 μm) B-F) Quantitative analysis of MMP2, MMP9, VCAM, MCP1 and IL-6 expression in 5 groups of mice (n = 6 per group). G) Representative western blot analysis of MMP2, MMP9, VCAM, MCP1, and IL-6 expression in abdominal aortas of 5 groups of mice and their quantitative analysis H-L) (n = 6 per group). M) Representative zymography of MMP2 and MMP9 activity in abdominal aortas of 5 groups of mice and their quantitative analyses (N-O) (n = 6 per group). *p <0.05, **p <0.01, ***p <0.001, One-way ANOVA followed by Tukey test for post hoc comparison, mean ± SEM.

Article Snippet: In the first part of the in vitro experiments, to examine the dose-response relation between formoterol treatment and VSMC senescence, VSMC were divided into 6 groups, which were treated with vehicle (PBS), 5IU mL−1 EPO, 5IU mL−1 EPO+0.01 nmol mL−1 formoterol (S2020, Selleck, USA), 5IU mL−1 EPO+0.1 nmol mL−1 formoterol, 5IU mL−1 EPO+1 nmol mL−1 formoterol and 5IU mL−1 EPO+10 nmol mL−1 formoterol, respectively.

Techniques: Expressing, Immunohistochemical staining, Staining, Western Blot, Zymography, Activity Assay, Comparison

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Medium-Dose Formoterol Attenuated Abdominal Aortic Aneurysm Induced by EPO via β2AR/cAMP/SIRT1 Pathway.

doi: 10.1002/advs.202306232

Figure Lengend Snippet: Figure 4. Effect of formoterol (FORM) on EPO-induced apoptosis of abdominal aortic tissues. A) Representative immunohistochemical staining of the abdominal aortic sections for 𝛼SMA in 5 groups of mice (scale bars = 10 μm) B) Quantitative analysis of 𝛼SMA expression in the abdominal aortic sections of 5 groups of mice (n = 6 per group) C) Representative western blot analysis of 𝛼SMA expression in abdominal aortas of 5 groups of mice and quantitative analysis (D). E) Representative TUNEL images in abdominal aortas of 5 groups of mice (scale bars = 25 μm) and quantitative analysis (F). G) Representative western blot analysis of BAX and BCL2 in abdominal aortas of 5 groups of mice and quantitative analysis (H) and (I) (n = 6 per group). *p <0.05, **p <0.01, ***p <0.001, One-way ANOVA followed by Tukey test for post hoc comparison, mean ± SEM.

Article Snippet: In the first part of the in vitro experiments, to examine the dose-response relation between formoterol treatment and VSMC senescence, VSMC were divided into 6 groups, which were treated with vehicle (PBS), 5IU mL−1 EPO, 5IU mL−1 EPO+0.01 nmol mL−1 formoterol (S2020, Selleck, USA), 5IU mL−1 EPO+0.1 nmol mL−1 formoterol, 5IU mL−1 EPO+1 nmol mL−1 formoterol and 5IU mL−1 EPO+10 nmol mL−1 formoterol, respectively.

Techniques: Immunohistochemical staining, Staining, Expressing, Western Blot, TUNEL Assay, Comparison

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Medium-Dose Formoterol Attenuated Abdominal Aortic Aneurysm Induced by EPO via β2AR/cAMP/SIRT1 Pathway.

doi: 10.1002/advs.202306232

Figure Lengend Snippet: Figure 5. Formoterol (FORM) suppressed EPO-induced VSMC senescence in mouse aorta. A) GO enrichment for the intersection genes. B) KEGG enrichment for the intersection genes. C) Representative immunofluorescent analysis of the abdominal aortic sections for detecting the colocalization (yellow particles) of 𝛾H2AX (red particles) and 𝛼SMA (specific for SMCs, green particles) in ApoE−/−mice receiving vehicle, EPO and EPO+ medium- dose formoterol treatment, respectively (scale bars = 10 μm). D) Quantitative analysis of 𝛾H2AX/𝛼SMC colocalization in (C) (n = 6 per group). E) Representative immunofluorescent analysis of the abdominal aortic sections for detecting the colocalization (yellow particles) of SIRT1 (green particles) and 𝛼SMA (specific for SMCs, red particles) in ApoE−/−mice receiving vehicle, EPO and EPO+medium-dose formoterol treatment, respectively. (scale bars = 10 μm). F) Quantitative analysis of 𝛾H2AX/𝛼SMC colocalization in (E) (n = 6 per group). *p <0.05, **p <0.01, ***p <0.001, One-way ANOVA followed by Tukey test for post hoc comparison, mean ± SEM.

Article Snippet: In the first part of the in vitro experiments, to examine the dose-response relation between formoterol treatment and VSMC senescence, VSMC were divided into 6 groups, which were treated with vehicle (PBS), 5IU mL−1 EPO, 5IU mL−1 EPO+0.01 nmol mL−1 formoterol (S2020, Selleck, USA), 5IU mL−1 EPO+0.1 nmol mL−1 formoterol, 5IU mL−1 EPO+1 nmol mL−1 formoterol and 5IU mL−1 EPO+10 nmol mL−1 formoterol, respectively.

Techniques: Comparison

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Medium-Dose Formoterol Attenuated Abdominal Aortic Aneurysm Induced by EPO via β2AR/cAMP/SIRT1 Pathway.

doi: 10.1002/advs.202306232

Figure Lengend Snippet: Figure 7. cAMP-regulated the effect of formoterol on VSMC senescence. A) Quantification of intracellular cAMP levels in VSMC treated with mAC inhibitor SQ22536 (80 μM) or control solution DMSO receiving vehicle, EPO and EPO+0.1 nmol mL−1 formoterol, respectively. B) Representative images of SA-𝛽-gal staining of VSMC treated with mAC inhibitor SQ22536 (80 μM) or control solution DMSO receiving vehicle, EPO and EPO+0.1 nmol mL−1 formoterol, respectively. (scale bars = 10 μm). C) Quantitative analysis of SA-𝛽-gal staining in (B) (n = 6 per group). D) Representative western blot assay of SIRT1, P21 in VSMC treated with mAC inhibitor SQ22536 (80 μM) or control solution DMSO receiving vehicle, EPO and EPO+0.1 nmol mL−1 formoterol, respectively. E-F) Quantitative analysis of protein expression of SIRT1, P21 protein levels by western blot in (D), (n = 6 per group). G) Representative images of immunostaining of SIRT1 in VSMC treated with mAC inhibitor SQ22536 (80 μM) or control solution DMSO receiving vehicle, EPO and EPO+0.1 nmol mL−1 formoterol, respectively. H) Quantitative analysis of colocalization of dapi/SIRT1 in (G) (n = 6 per group). *p < 0.05, **p <0.01, ***p <0.001, Two-way ANOVA followed by Tukey test for post hoc comparison, mean ± SEM.

Article Snippet: In the first part of the in vitro experiments, to examine the dose-response relation between formoterol treatment and VSMC senescence, VSMC were divided into 6 groups, which were treated with vehicle (PBS), 5IU mL−1 EPO, 5IU mL−1 EPO+0.01 nmol mL−1 formoterol (S2020, Selleck, USA), 5IU mL−1 EPO+0.1 nmol mL−1 formoterol, 5IU mL−1 EPO+1 nmol mL−1 formoterol and 5IU mL−1 EPO+10 nmol mL−1 formoterol, respectively.

Techniques: Control, Staining, Western Blot, Expressing, Immunostaining, Comparison

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Medium-Dose Formoterol Attenuated Abdominal Aortic Aneurysm Induced by EPO via β2AR/cAMP/SIRT1 Pathway.

doi: 10.1002/advs.202306232

Figure Lengend Snippet: Figure 9. Schematic diagram showing the mechanism of therapeutic effects of medium-dose formoterol on EPO-induced AAA. Formoterol binds to 𝛽2AR and activates cAMP, which increases SIRT1 protein expression, leading to suppressed VSMC senescence induced by EPO. In contrast, SIRT1 is downregulated by EPO via activation of CBL, resulting in aggravated VSMC senescence. Thus, medium-dose formoterol attenuated EPO-induced AAA via 𝛽2AR/cAMP/SIRT1 pathways, which provides a promising medication for the treatment of AAA.

Article Snippet: In the first part of the in vitro experiments, to examine the dose-response relation between formoterol treatment and VSMC senescence, VSMC were divided into 6 groups, which were treated with vehicle (PBS), 5IU mL−1 EPO, 5IU mL−1 EPO+0.01 nmol mL−1 formoterol (S2020, Selleck, USA), 5IU mL−1 EPO+0.1 nmol mL−1 formoterol, 5IU mL−1 EPO+1 nmol mL−1 formoterol and 5IU mL−1 EPO+10 nmol mL−1 formoterol, respectively.

Techniques: Expressing, Activation Assay