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. 2016 Jan;89(1):94–104. doi: 10.1124/mol.115.100339

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Copyright © 2015 by The American Society for Pharmacology and Experimental Therapeutics

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Fig. 4. — FR900359, but not P4pal-10, can inhibit airway contraction in human precision cut lung slices. (A) One hundred nanomolars and 1 μM FR900359 inhibited carbachol-promoted airway contraction. The data are represented by the mean ± S.E.M. of four independent experiments and fitted to a sigmoidal dose-response model. (B) P4pal-10 and P4pal-10 Scr had no effect on airway contraction in response to carbachol. The data are represented by the mean ± S.E.M. of four independent experiments and fitted to a sigmoidal dose-response model. The no inhibitor curve is derived from the same data set in both (A and B). (C) One hundred nanomolars or 1 μM of FR900359 shifted the estimated EC₅₀ value of carbachol-promoted airway contraction by 2- and 11-fold, respectively. P4pal-10 and P4pal-10 Scr did not modulate carbachol responsiveness. The data are represented by the mean ± S.E.M. of four independent experiments. ns, not significant. ***P < 0.001 versus vehicle-treated tissue using a two-way unpaired t test. (D) One hundred nanomolars and 1 μM of FR900359 effectively inhibited histamine-promoted airway contraction. The data are represented by the mean ± S.E.M. of three independent experiments and fitted to a sigmoidal dose-response model. (E) P4pal-10 partially inhibited airway contraction in response to histamine. The data are represented by the mean ± S.E.M. of three independent experiments and fitted to a sigmoidal dose-response model. The no inhibitor curve is derived from the same data set in both (D and E). (F) P4pal-10 shifted the estimated EC₅₀ value of histamine-promoted airway contraction by 8-fold, whereas 100 nM and 1 μM FR900359 shifted the EC₅₀ > 50-fold. The data are represented by the mean ± S.E.M. of three independent experiments. *P < 0.05 and **P < 0.01 versus vehicle-treated tissue using a two-way unpaired t test. (G) Magnetic twisting cytometry analysis of isolated ASM dynamics shows that pretreatment with 3 μM P4pal-10 (n = 232) significantly inhibited carbachol-promoted single-cell contraction as compared with no inhibitor/vehicle treatment (n = 331). P4pal-10 Scr did not exhibit similar efficacy (n = 266). The data sets are normalized to no inhibitor/vehicle treatment. ***P < 0.001 compared with vehicle-treated cells using a one-way analysis of variance (ANOVA) analysis. (H) Magnetic twisting cytometry analysis of isolated ASM dynamics shows that pretreatment with 3 μM P4pal-10 (n = 867) significantly inhibited histamine-promoted single-cell contraction as compared with no inhibitor/vehicle treatment (n = 692). P4pal-10 Scr did not exhibit similar efficacy (n = 486). The data sets are normalized to no inhibitor/vehicle treatment. ***P < 0.001 compared with vehicle-treated cells using a one-way ANOVA analysis. (I) Magnetic twisting cytometry analysis of isolated ASM dynamics shows that pretreatment with 3 μM P4pal-10 (n = 418) or P4pal-10 Scr (n = 292) did not modulate isoproterenol-promoted single-cell relaxation as compared with no inhibitor/vehicle treatment (n = 569). The data sets are normalized to no inhibitor/vehicle treatment. ns, not significant compared with vehicle-treated cells using a one-way ANOVA analysis.