In human airway smooth muscle cells, osthole increased VASP phosphorylation. Suppression by PKI and H89 supports PKA-dependent signaling. [2]
Inhibition of reconstituted mouse TRPV3 channels
Biochemical assay · Wild-type mouse TRPV3 incorporated into planar lipid bilayers; single-channel recordings at +80 mV. [1]
Osthole reduced 2-APB-evoked channel opening in a cell-free system.
Literature-Reported Treatment Conditions
- Treatment concentration
- 5, 25 or 50 μM osthole
- Exposure duration
- Exposure duration not specified for each concentration in Figure 1.
- Method
- Single-channel recording in planar lipid bilayers.
- Co-treatment
- 5 μM 2-APB used to activate channels; comparison with 2-APB alone.
- Reported limits
- Bilayer-specific osthole vehicle composition and exposure times were not specified in the selected recording methods; DMSO reported for structural sample preparation is not assigned to this assay.
Methods and source locations
Compound reported in the paper: Osthole; Compound label in the paper: osthole
Evidence concerns the paper material, not a tested Adooq batch.
Results: Functional characterization and cryo-EM; Figure 1A-B and caption; Materials and Methods: Planar lipid bilayer recordings
TRPV3 open probability at +80 mV: Planar lipid bilayer recording with 5 μM 2-APB; one-way ANOVA followed by Fisher least significant difference test. — Results: Functional characterization and cryo-EM, first paragraph; Figure 1A-B/caption
Inhibition of recombinant active PDE4D5
Biochemical assay · Recombinant c-Flag-tagged PDE4D5-S126D; S126D-D556A enzymatically dead mutant and mock controls. [2]
Osthole concentration-dependently inhibited the constitutively active PDE4D5 mutant; the comparison is not an IC50 determination.
Literature-Reported Treatment Conditions
- Treatment concentration
- 0.1, 1 and 10 μM osthole
- Exposure duration
- 20 min preincubation; 1 hour after cAMP addition.
- Method
- PDELight HTS cAMP phosphodiesterase assay at room temperature; 2 μM cAMP.
- Co-treatment
- Rolipram 10 μM was a separate positive comparator, not combined with osthole.
- Reported limits
- Assay-specific vehicle composition not specified in the retrieved PDE assay method.
Methods and source locations
Compound reported in the paper: Osthole; Compound label in the paper: osthole
Evidence concerns Ark Pharm material, not a tested Adooq batch.
Results: Osthole inhibits PDE4D5 activity in vitro; Figure 8A-B/caption; Methods: In vitro assay of PDE activity; Reagents and cells; Expression of PDE4D5 protein in mammalian cells
PDE4D5-S126D cAMP phosphodiesterase activity: PDELight assay; Figure 8A-B: means ± SEM, n = 3 separate experiments in technical duplicate; ANOVA with Bonferroni correction. — Results: Osthole inhibits PDE4D5 activity in vitro; Figure 8A-B and caption
PKA-dependent VASP phosphorylation in human airway smooth muscle cells
Cell-based study · Three human airway smooth muscle cell lines derived from nonasthmatic donor lung tissue; passages 3–8. [2]
Osthole increased VASP Ser157 phosphorylation; PKI and H89 suppressed the response.
Literature-Reported Treatment Conditions
- Treatment concentration
- 0.1, 0.3, 1, 3 and 10 μM osthole; inhibitor experiment: 3 μM osthole.
- Exposure duration
- 15 min osthole stimulation in Figure 3C.
- Method
- Western blot; phospho-VASP signal normalized to total VASP.
- Co-treatment
- Separate inhibitor conditions used myristoylated PKI (10 μM) or H89 (10 μM), with vehicle controls.
- Reported limits
- Figure 3D inhibitor pretreatment interval and assay vehicle composition not specified in its caption or shared Western blot method; do not infer from other panels.
Methods and source locations
Compound reported in the paper: Osthole; Compound label in the paper: osthole
Evidence concerns Ark Pharm material, not a tested Adooq batch.
Results: Osthole stimulates cAMP accumulation to activate PKA in human ASM cells; Figure 3C-D/caption; Methods: Reagents and cells; Western blot
VASP Ser157 phosphorylation: Western blot and densitometry; means ± SEM, n = 3 different human ASM cell lines; experiments repeated multiple times (exact repeat count unspecified). — Results: Osthole stimulates cAMP accumulation to activate PKA in human ASM cells; Figure 3C-D/caption
