Journal: British Journal of Pharmacology

Article Title: Preserved arterial vasodilatation via endothelial protease-activated receptor-2 in obese type 2 diabetic mice

doi: 10.1111/j.1476-5381.2011.01356.x

Figure Lengend Snippet: Effects of an AC inhibitor (SQ22536) and PGI 2 receptor antagonist (CAY10441) on 2-furoyl-LIGRLO-amide (2fly)-induced relaxations of db/db and C57BL/6J (C57) mesenteric arteries. Values are the mean ± SE ( n = number of mice) for 2fly-induced relaxations of C57 and db/db second order mesenteric arteries contracted submaximally by U46619 in the absence or presence of (A, B) SQ22536 (100 µM) (C, D) CAY10441 (0.3 µM). Arteries were exposed to inhibitors for 30 min prior to U46619. There were no significant effects of inhibitors or strain on relaxation responses ( P > 0.05, 2-way anova ).

Article Snippet: Other sources included: Bachem (Torrance, CA, USA), charybdotoxin; Cayman Chemicals (Ann Arbor, MI, USA), CAY10441, SC560, SC58125 and SQ22536; Peptides International (Louisville, KT, USA), 2-furoyl-leu-ile-gly-arg-leu-orn-amide (2fly); Tocris BioScience (Ellisville, MO, USA): charybdotoxin (ChTx), FR122047, NS398; and University of Calgary Peptide Synthesis (Calgary, Alberta, Canada), 2-furoyl-orn-leu-arg-gly-ile-leu-amide.

Techniques:

Journal: Scientific Reports

Article Title: Adenosine inhibits TNFα-induced MMP-3 production in MH7A rheumatoid arthritis synoviocytes via A 2A receptor signaling

doi: 10.1038/s41598-022-10012-6

Figure Lengend Snippet: Effect of cAMP analog dbcAMP and the adenylate cyclase inhibitor SQ22536 on TNFα-induced enhancement of MMP-3 production. Activation of cAMP signaling suppressed TNFα-induced MMP-3 production ( a ). MH7A cells were incubated for 24 h in TNFα (25 pg/ml) with or without the indicated concentration of dbcAMP, and MMP-3 concentration measured in the culture medium. In some experiments, the cells were also pretreated for 30 min with 0.1 mM of the adenylate cyclase inhibitor SQ22536 ( b ). MMP-3 was then measured in the culture medium. The suppressive effect of HENECA on TNFα-induced MMP-3 production was blocked by SQ22536 pretreatment. Experiments were repeated three times and data are presented as mean ± SD. ★ p < 0.01 vs. TNFα alone.

Article Snippet: The adenylate cyclase inhibitor 9-(tetrahydrofuran-2-yl)-9 h-purin-6-amine (SQ22536) was purchased from TCI (Tokyo, Japan).

Techniques: Activation Assay, Incubation, Concentration Assay

Journal: Foods

Article Title: Sudachitin and Nobiletin Stimulate Lipolysis via Activation of the cAMP/PKA/HSL Pathway in 3T3-L1 Adipocytes

doi: 10.3390/foods12101947

Figure Lengend Snippet: Inhibition of sudachitin- and nobiletin-induced lipolysis using AC and PKA inhibitors. Lipolysis is assessed by the glycerol content in the medium containing 3T3-L1 adipocytes treated with 0.05% DMSO, SQ22536 (100 µM), or H-89 (20 µM) for 1 h and then treated with 0.05% DMSO, sudachitin, nobiletin (50 µM), or isoproterenol (10 µM) for 3 h. Data are expressed as means ± SEM ( n = 4). * p < 0.05 and ** p < 0.01 versus cells treated with DMSO and without inhibitors. †† p < 0.01 versus cells treated with the corresponding reagent and without inhibitors. AC: adenylate cyclase; DMSO: dimethyl sulfoxide; PKA: protein kinase A; SEM: standard error of the mean.

Article Snippet: The structures of sudachitin and nobiletin are shown in A. Cyclic AMP ELISA Kit, H-89, isoproterenol, and SQ22536 were purchased from Cayman Chemical (Ann Arbor, MI, USA). cOmplete Protease Inhibitor Cocktail was purchased from Roche Diagnostics (Indianapolis, IN, USA).

Techniques: Inhibition

Journal: Foods

Article Title: Sudachitin and Nobiletin Stimulate Lipolysis via Activation of the cAMP/PKA/HSL Pathway in 3T3-L1 Adipocytes

doi: 10.3390/foods12101947

Figure Lengend Snippet: Inhibition of sudachitin- and nobiletin-induced phosphorylation of PKA substrates and HSL by AC and PKA inhibitors. ( A , C ) Protein levels of PKA substrates phosphorylated, HSL phosphorylated at Ser563, HSL phosphorylated at Ser660, total HSL, and GAPDH. 3T3-L1 adipocytes are treated with 0.05% DMSO, SQ22536 (100 µM) ( A ), or H-89 (20 µM) ( C ) before treatment with 0.05% DMSO, sudachitin, or nobiletin (30 µM). ( B , D ) Densitometric ratios of phosphorylated PKA substrates/GAPDH and phosphorylated protein/total protein. Data are expressed as means ± SEM ( n = 4). * p < 0.05 and ** p < 0.01 versus cells treated with DMSO and without inhibitors. † p < 0.05 and †† p < 0.01 versus cells treated with the corresponding reagent and without inhibitors. DMSO: dimethyl sulfoxide; GAPDH: glyceraldehyde 3-phosphate dehydrogenase; HSL: hormone-sensitive lipase; PKA: protein kinase A; SEM: standard error of the mean.

Article Snippet: The structures of sudachitin and nobiletin are shown in A. Cyclic AMP ELISA Kit, H-89, isoproterenol, and SQ22536 were purchased from Cayman Chemical (Ann Arbor, MI, USA). cOmplete Protease Inhibitor Cocktail was purchased from Roche Diagnostics (Indianapolis, IN, USA).

Techniques: Inhibition

Journal: Nature Communications

Article Title: Cystic fibrosis swine fail to secrete airway surface liquid in response to inhalation of pathogens

doi: 10.1038/s41467-017-00835-7

Figure Lengend Snippet: CF tissues fail to produce basal ASL secretion. a CFTR −/− swine (CF, n = 9 from six animals) display lower basal ASL secretion than wild-type in vivo ( n = 10 from seven animals, mean ± SEM, P = 0.001, Mann–Whitney test). b Wild-type ex vivo preparations with surgically removed submucosal glands (SMG) produced lower basal ASL ( n = 12 with SMG from six preparations and n = 13 without SMG from six preparations, P < 0.0001, Mann–Whitney test). c Wild-type ex vivo preparations incubated with CFTRinh172 (100 µM) produced lower basal ASL than non-treated preparations ( n = 6 from four preparations for control, and n = 14 from five preparations for CFTRinh172, P < 0.0001, Mann–Whitney test). d Topical application of lidocaine (Lido) had a small effect on basal ASL secretion. Simultaneous treatment with the CFTR inhibitor (Lido + inh172) completely blocked basal ASL secretion ( n = 13 control from five preparations, n = 21 Lido from eight preparations, and n = 16 Lido + inh172 from seven preparations; P < 0.0001, F (2, 47) = 59.35, ANOVA and Tukey’s multiple comparison test). e Tetrodotoxin (1 µM, TTX) treatment had no effect on basal ASL secretion ( n = 18 control from eight preparations, and n = 11 TTX from four preparations, P = 0.58, Mann–Whitney test). Atropine had no effect on basal ASL secretion in f ex vivo preparations (10 µM, n = 8 without atropine from three preparations and n = 8 with atropine from four preparations, P = 0.76, t = 0.30, df = 14, Student’s t test) or g in vivo wild-type swine (treated with 0.04 mg/kg IM 2–10 min following induction of anesthesia, n = 8 without Atropine from six animals, and n = 10 with Atropine from seven animals, P = 0.75, t = 0.32, df = 16, Student’s t test). h Treatment with SQ22536 (0.5 mM) blocked ASL secretion ( n = 10 control from four preparations and n = 11 SQ22536 from four preparations; P = 0.0003, Mann–Whitney test). Data is presented as mean ± SEM and within each panel the columns labeled with an asterisk differ significantly

Article Snippet: CFTRinh-172, SQ22536, and flagellin from Salmonella typhimurium were obtained from Cedarlane Labs (Burlington, ON, CA).

Techniques: In Vivo, MANN-WHITNEY, Ex Vivo, Produced, Incubation, Control, Comparison, Labeling