Background
Methods
Patients and human bronchus samples
Tissue preparation for organ bath studies
Low-frequency, low-force cyclic stretching
Functional assays
Drugs
Analysis of the organ bath fluid
RNA isolation, and reverse transcription quantitative polymerase chain reaction (RT-qPCR array analysis)
Data analysis
Results
Effect of cyclic stretching on bronchial basal tone and responsiveness
Effect of epithelium removal on the impact of cyclic stretching
Cyclic stretch effect on the basal tone during stretching period, difference between control and stretch group | Effect of pretreatment | Effect size pretreatment | |||
---|---|---|---|---|---|
n
| in absence of pretreatment (g) | in presence of pretreatment (g) | (g) | |d| [95% CI] | |
Epithelium removal | 13 | 0.76 ± 0.08
a
| 0.53 ± 0.10
a
| −0.23 ± 0.09
d
| −0.70 [−1.48 − 0.10] |
Indomethacin (1 μM) | 8 | 1.14 ± 0.2
b
| 1.14 ± 0.18
a
| 0.01 ± 0.10 | 0.00 [−1.01 − 1.01] |
MK476 (1 μM) | 7 | 1.22 ± 0.2
b
| 0.89 ± 0.14
a
| −0.33 ± 0.12
d
| −0.68 [−1.70 − 0.44] |
L-NAME (1 mM) | 14 | 0.61 ± 0.15
b
| 1.18 ± 0.15
a
| 0.57 ± 0.14
e
| 1.02 [0.20 − 1.77] |
Nicardipine (10 μM) | 10 | 0.65 ± 0.12
a
| 1.03 ± 0.18
a
| 0.38 ± 0.18 | 0.78 [−0.16 − 1.66] |
Tetrodotoxin (1 μM) | 7 | 1.07 ± 0.19
b
| 1.29 ± 0.28
b
| 0.22 ± 0.16 | 0.35 [−0.73 − 1.38] |
Gadolinium (0.1 mM) | 8 | 1.18 ± 0.20
a
| 1.24 ± 0.15
a
| 0.06 ± 0.18 | 0.12 [−0.87 − 1.10] |
Tertiapine (10 μM) | 9 | 0.51 ± 0.13
b
| −0.74 ± 0.12
a
| −1.25 ± 0.17
f
| −3.32 [−4.50− −1.85] |
Capsazepine (1 μM) | 8 | 1.19 ± 0.14
a
| 0.92 ± 0.20
b
| −0.26 ± 0.23 | −0.67 [−1.31− −0.01] |
Calphostine C (0.1 μM) | 9 | 1.10 ± 0.23
a
| 1.12 ± 0.25
b
| 0.02 ± 0.29 | 0.03 [−0.90 − 0.95] |
SB203580 (0.1 μM) | 10 | 1.11 ± 0.27
b
| 0.84 ± 0.42 | −0.27 ± 0.37 | −0.24 [−1.11 − 0.65] |
Y27632 (1 μM) | 11 | 1.10 ± 0.25
b
| −0.75 ± 0.30
c
| −1.85 ± 0.50
e
| −2.03 [−2.96– − 0.93] |
Cyclic stretch effect on post-stretch response, difference between control and stretch group | Effect of pretreatment | Effect size pretreatment | |||
---|---|---|---|---|---|
n
| in absence of pretreatment (g) | in presence of pretreatment (g) | (g) | |d| [95% CI] | |
Epithelium removal | 13 | 0.24 ± 0.03
a
| 0.15 ± 0.03
a
| −0.09 ± 0.03
d
| −3.00 [−4.01– − 1.80] |
Indomethacin (1 μ M) | 8 | 0.36 ± 0.07
b
| 0.29 ± 0.07
b
| −0.07 ± 0.11 | −0.37 [−1.34–0.64] |
MK476 (1 μM) | 7 | 0.35 ± 0.08
b
| 0.23 ± 0.06
c
| −0.12 ± 0.10 | −0.64 [−1.67–0.47] |
L-NAME (1 mM) | 14 | 0.29 ± 0.03
a
| 0.21 ± 0.03
a
| −0.08 ± 0.02
d
| −0.73 [−1.47–0.06] |
Nicardipine (10 μM) | 10 | 0.17 ± 0.03
a
| 0.12 ± 0.08
a
| −0.05 ± 0.07 | −0.27 [−1.13–0.63] |
Tetrodotoxin (1 μM) | 7 | 0.37 ± 0.08
b
| 0.29 ± 0.06
b
| −0.07 ± 0.10 | −0.43 [−1.46–0.66] |
Gadolinium (0.1 mM) | 8 | 0.36 ± 0.07
b
| 0.26 ± 0.03
a
| −0.10 ± 0.08 | −0.69 [−1.65 − 0.36] |
Tertiapine (10 μM) | 9 | 0.23 ± 0.06
b
| 0.27 ± 0.04
a
| 0.04 ± 0.08 | 0.26 [−0.68–1.18] |
Capsazepine (1 μM) | 8 | 0.29 ± 0.06
b
| 0.37 ± 0.05
a
| 0.08 ± 0.06 | 0.74 [−0.31–1.71] |
Calphostine C (0.1 μM) | 9 | 0.21 ± 0.03
a
| 0.26 ± 0.02
a
| 0.05 ± 0.05 | 0.65 [−0.32–1.57] |
SB203580 (0.1 μM) | 10 | 0.29 ± 0.05
a
| 0.15 ± 0.19 | −0.14 ± 0.22 | −0.32 [−1.19–0.58] |
Y27632 (1 μM) | 11 | 0.28 ± 0.05
a
| 0.33 ± 0.07
b
| 0.05 ± 0.08 | 0.25 [−0.60–1.08] |
Cyclic stretch effect on ΔEmax (ACh) difference between control and stretch group | Effect of pretreatment | Effect size pretreatment | |||
---|---|---|---|---|---|
n
| in absence of pretreatment (g) | in presence of pretreatment (g) | (g) | |d| [95% CI] | |
Epithelium removal | 13 | −0.01 ± 0.15 | 0.27 ± 0.14 | 0.27 ± 0.12
a
| 0.54 [−0.26–1.29] |
Indomethacin (1 μM) | 8 | 0.12 ± 0.25 | 0.19 ± 0.11 | 0.07 ± 0.25 | 0.25 [−0.75–1.22] |
MK476 (1 μM) | 7 | 0.15 ± 0.29 | 0.51 ± 0.23 | 0.37 ± 0.28 | 0.53 [−0.57–1.55] |
L-NAME (1 mM) | 14 | −0.01 ± 0.07 | 0.26 ± 0.14 | 0.25 ± 0.11
a
| 0.66 [−0.12–1.40] |
Nicardipine (10 μM) | 10 | 0.25 ± 0.18 | 1.02 ± 0.44
b
| 0.76 ± 0.43 | 0.73 [−0.21–1.60] |
Tetrodotoxin (1 μM) | 7 | 0.16 ± 0.25 | −0.02 ± 0.12 | −0.18 ± 0.30 | −0.35 [−1.38–0.73] |
Gadolinium (0.1 mM) | 8 | 0.11 ± 0.22 | 0.39 ± 0.19 | 0.28 ± 0.28 | 0.49 [−0.53–1.45] |
Tertiapine (10 μM) | 9 | −0.02 ± 0.08 | 0.25 ± 0.10
b
| 0.27 ± 0.18 | 0.95 [−0.03–1.92] |
Capsazepine (1 μM) | 8 | 0.36 ± 0.26 | 0.29 ± 0.19 | −0.07 ± 0.26 | −0.11 [−1.08–0.88] |
Calphostine C (0.1 μM) | 9 | 0.00 ± 0.28 | 0.18 ± 0.22 | 0.18 ± 0.22 | 0.24 [−0.70–1.15] |
SB203580 (0.1 μM) | 10 | −0.22 ± 0.21 | −0.25 ± 0.16 | −0.03 ± 0.25 | −0.05 [−0.93–0.83] |
Y27632 (1 μM) | 11 | −0.13 ± 0.22 | 0.21 ± 0.08
b
| 0.33 ± 0.21 | 0.63 [−0.25–1.46] |
Cyclic stretch effect on Δ (−log EC50), difference between control and stretch group | Effect of pretreatment | Effect size pretreatment | |||
---|---|---|---|---|---|
n
| in absence of pretreatment (log unit) | in presence of pretreatment (log unit) | (log unit) | |d| [95% CI] | |
Epithelium removal | 13 | 0.48 ± 0.08
a
| 0.50 ± 0.14
b
| 0.02 ± 0.13 | 0.05 [−0.70–0.80] |
Indomethacin (1 μM) | 8 | 0.38 ± 0.07
a
| 0.32 ± 0.13
b
| −0.05 ± 0.12 | −0.23 [−1.20–0.76] |
MK476 (1 μM) | 7 | 0.41 ± 0.07
c
| 0.26 ± 0.23 | −0.15 ± 0.24 | −0.34 [−1.37–0.74] |
L-NAME (1 mM) | 14 | 0.30 ± 0.07
a
| 0.44 ± 0.05
a
| 0.14 ± 0.05 | 0.54 [−0.23–1.28] |
Nicardipine (10 μM) | 10 | 0.57 ± 0.10
a
| 0.11 ± 0.12 | −0.45 ± 0.17
d
| −1.26 [−2.16– − 0.25] |
Tetrodotoxin (1 μM) | 7 | 0.25 ± 0.05
c
| 0.38 ± 0.04
a
| 0.13 ± 0.06 | 1.12 [−0.07–2.16] |
Gadolinium (0.1 mM) | 8 | 0.27 ± 0.05
b
| 0.49 ± 0.08
a
| 0.22 ± 0.08
d
| 1.19 [0.07–2.18] |
Tertiapine (10 μM) | 9 | 0.40 ± 0.13
b
| 0.41 ± 0.09
c
| 0.03 ± 0.19 | 0.03 [−0.90–0.95] |
Capsazepine (1 μM) | 8 | 0.40 ± 0.11
c
| 0.54 ± 0.12
c
| 0.14 ± 0.15 | 0.41 [−0.60–1.38] |
Calphostine C (0.1 μM) | 9 | 0.26 ± 0.04
a
| 0.24 ± 0.19 | −0.01 ± 0.20 | −0.04 [−0.80–0.88] |
SB203580 (0.1 μM) | 10 | 0.37 ± 0.07
a
| 0.36 ± 0.10 | −0.01 ± 0.13 | −0.04 [−0.91–0.84] |
Y27632 (1 μM) | 11 | 0.37 ± 0.08
a
| 0.37 ± 0.16
b
| −0.01 ± 0.16 | −0.12 [−0.95–0.72] |