Effect and application of saikoside B1 in preparation of inhibitors of Hedgehog signal pathway
A technology of saikosaponin and signaling pathways, applied in the field of medicine, can solve the problems of complicated pathogenesis, no good solution for prevention and treatment, etc., and achieve the effect of expanding the application field
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0032] Example 1: Experiment of specific inhibitory effect of SSB1 on Hedgehog signaling pathway in LightII cells in vitro
[0033] 1.1 Dual luciferase reporter gene test
[0034]LightII (American Type Culture Collecton) cells were treated with Dulbecco's modified Eagle's medium (DMEM) (GIBCOB) containing 10% (v / v) newborn bovine serum (FBS, GIBCO), penicillin / streptomycin (Hyclone), 37°C, 5 Cultured at constant temperature with %CO2, seeded in 96-well plate with 8×103 cells / well, replaced with medium containing 1% FBS after 24 hours, SAG (100nM) or SHHN conditioned medium (SHHN-CM) and formulated with different concentrations of The cells were treated with SSB1 (Feng Biological Technology Ltd, Shanghai6), and a blank control group and a positive control group were set at the same time. After incubation at a constant temperature of 37°C and 5% CO2 for 36 hours, Gli1- For the detection of the relative activity of luciferase, three replicate wells were set up for each concentra...
Embodiment 2
[0038] Example 2: Experimental research on the target of SSB1 on the Hedgehog signaling pathway
[0039] 2.1 Inhibition of SSB1 on the activity of Gli luciferase activated after transfection with Smoothened
[0040] LightII cells were cultured in DMEM containing 10% (v) FBS, penicillin / streptomycin, 37°C, 5% CO2 at a constant temperature, seeded in 96-well plates at 8×103 cells / well, and transfected with lipo2000 respectively after 24 hours. Transfected with SmoD473H and Gli2△N plasmids, 36 hours after transfection, replaced with DMEM medium containing 1% FBS, treated cells with different concentrations of SSB1, incubated at 37°C, 5% CO2 for 36 hours, and treated with double fluorescein Enzyme reporter system was used to measure the relative activity of Gli-luciferase (Promega). Three replicate wells were set up for each concentration in this experiment and repeated at least three times;
[0041] 2.2 Inhibition of SSB1 on the activity of Gli luciferase activated after virus i...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More