- Natural product: Cat's-claw (Uncaria tomentosa)
- Associated study: Identification of PXR activators from Uncaria tomentosa (Cat’s Claw)
In Vitro Nuclear Receptor Induction Experiment
Mitraphylline Effect on PXR Activation
Induction was detected. Cutoff used —
Not available
- Pregnane X receptor (PXR)
Cell system PXR-transfected hepatic cells
Results
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
DPX2 cells (~ 60,000 cells/well) were seeded on 96-well plate (clear bottom and white wall). After 24 h incubation in 37 °C incubator (5% CO2), cells were treated with herbal extracts or chemicals in dosing medium for another 24 h. Each chemical tested was dissolved in DMSO to prepare a stock solution (10 mM). The final concentration of DMSO was 1‰ in control and treatment groups. Cell viability was detected by replacing the medium with 100 µl of CellTiter-FluroTM reagent, incubating for 1 h and measuring fluorescence (RFU) with an excitation wavelength of 390 nm and emission wavelength of 505 nm. Afterward, 100 µl ONE-GloTM assay reagent was added into each well. Plate was agitated carefully, and then luminescence (RLU) was detected 5 min later. The efficacy of PXR activation was calculated by the ratio of RLU/RFU for each well. Both RLU and RFU were measured by BioTekTM Synergy TM H1 Hybrid Reader. The half maximal effective concentration (EC50) was determined from the dose-response curve using GraphPad prism.
Protein expression
60,000 cells/well seeded on 96-well plate
Yes
10 µM
24 hours
48 hours