Music W, Wei S, Liu G, Yu Z, Estell K, Yadav AK, Schwiebert LM, Matalon S. Postexposure administration of a 2-agonist decreases chlorine-induced airway hyperreactivity in mice. recognized improved concentrations of at 24C for 20 min. Supernatants were then collected and placed in polypropylene tubes. HPLC-grade water was added, making the final supernatant/water answer 25% organic. To isolate the compounds of interest partial purification of the 25% answer was performed on a Preppy apparatus put together with 500 mg C18 solid-phase extraction columns. The columns were conditioned with 5 ml of HPLC-grade methanol immediately followed by 2.5 ml of HPLC-grade water. The supernatant/water answer was then loaded onto the C18 column, and then washed with 2.5 ml of HPLC grade water followed by 1.5 HQ-415 ml of 40% methanol. Elutions of 1 1.5 ml of 60, 75, 85, and 100% methanol were collected in individual autosampler vials and stored at ?20C until analysis by mass spectrometry. LC/MS/MS analysis and quantification. HPLC/MS/MS methods previously described were used for each of the lipids analyzed here (7, 30). With the exception of the 2-acyl glycerol and and 0.05 or ** 0.01 vs. GSK1016790 by 1-way ANOVA Bonferroni post hoc analysis. 0.05 or ** 0.01 vs. HQ-415 GSK1016790 by 1-way ANOVA Bonferroni post hoc analysis. = 7C14/group. = 4C5/group. 0.01, *** 0.001, **** 0.0001 vs. respective controls. Table 1. TRPV4 inhibitor potencies assessed by TRPV4 ortholog transduction into HEK cells and hypotonicity assessed in BHK HQ-415 cells = 28)8.6 (= 14)GSK10167907.4 (= 4)8.2 (= 4)Hypotonicity7.6 (= 14)8.4 (= 7)RatGSK6347758.8 (= 5)8.4 (= 4)GSK10167908.5 (= 2)8.0 (= 2)MouseGSK6347758.7 (= 4)8.2 (= 6)GSK10167908.2 (= 4)7.7 (= 4)DogGSK6347757.8 (= 4)8.1 (= 6)GSK10167907.2 (= 4)7.9 (= 4)MonkeyGSK6347758.0 (= 4)8.5 (= 4)GSK10167907.6 (= 3)8.2 (= 4) Open in a separate window Table 2. TRPV4 inhibitor TRP selectivity profiles = 2) 4.6 (= 6)TRPA1 4.6 (= 2) 4.6 (= 2) 4.6 (= 4) 4.6 (= 4)TRPC3 4.6 (= 4) 4.6 (= 4) 4.6 (= 4) 4.6 (= 4)TRPC6 4.6 (= 4) 4.6 (= 4) 4.6 (= 6) 4.6 (= 4)TRPM5 4.6 (= 3) 4.6 (= 3) 4.6 (= 3) 4.6 (= 3)TRPM8 4.6 (= 2) Open in a separate window A single intraperitoneal injection of GSK2220691 (30 mg/kg) was administered 30 min after induction of injury by intratracheal administration of HCl (pH 1.5, 2 ml/kg), and inflammatory guidelines were analyzed after 5 h. BALF of GSK2220691-treated mice contained much smaller numbers of neutrophils and macrophages, and less MPO activity than BALF of vehicle-injected mice (Fig. 2, and ?and5).5). Multiplex peptide analyses of inflammatory cytokines and chemokines in BALF exposed that TRPV4 inhibition completely suppressed HCl-induced raises in key factors such as VEGF, keratinocyte-derived chemokine (KC; CXCL1), and HQ-415 granulocyte colony-stimulating element (GCSF) (Fig. 3and 0.05, ** 0.01, *** 0.001 vs. respective settings. and and and = 4C6/group. = Il1a 4C6/group. and = 4C6/group. and = 4C6/group. * 0.05, ** 0.01, *** 0.001 vs. air-exposed group; # 0.05, ## 0.01, ### 0.001 vs. chlorine-exposed group. Open in a separate windows Fig. 7. Effects of intramuscular TRPV4 inhibitor administration on markers of chlorine-induced swelling. = 5/group. 0.05, ### 0.001 vs. chlorine-exposed group. Anti-inflammatory effects of TRPV4 inhibitors in chlorine-injured mice. Chlorine-induced lung injury is definitely associated with strong pulmonary swelling driven by macrophages and neutrophils, levels of which were highly improved in BALF 24 h after exposure (Fig. 6, and and and and and ?and7and ?and8and ?and8= 12/group. 0.001, **** 0.0001 vs. respective controls. Open in a separate windows Fig. 8. Inflammatory cytokines and chemokines in BALF and serum, markers.
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