Isosakuranetin

Cell experiment:

B16 melanoma cells are cultured in 24-well plates at a density of 1×104 cells/well. After 24 h, the cells are incubated for 72 h in the presence of Isosakuranetin at the indicated concentrations. The culture medium is removed and replaced with 500 μL of fresh culture medium containing 10% sterile filtered MTT. After 3 h, the formazan crystals are dissolved in 500 μL/well isopropanol and absorbance is measured at 570 nm against 630 nm. Inhibition of proliferation (%) is expressed as the percentage of viable treated cells in comparison with control cells[1].

Animal experiment:

Male Sprague-Dawley rats weighing 250 to 300 g are habituated to the temperature, humidity, and lighting (12 h light/dark cycle, lights on at 7: 00 a.m.) controlled vivarium and singly housed for at least 1 week before the studies begin. The rotarod test is used to evaluate the potential motor-impairing effect of Isosakuranetin. For the time course studies, baseline measurement is immediately followed by an injection of Isosakuranetin, and then the paw-withdrawal threshold is measured every 15 min until the drug effect dissipates to a point that the paw withdrawal threshold is not significantly different from the predrug data or until 75 min passes[2].

References:

[1]. Drira R, et al. Isosakuranetin, a 4'-O-methylated flavonoid, stimulates melanogenesis in B16BL6 murine melanoma cells. Life Sci. 2015 Dec 15;143:43-9.
[2]. Jia S, et al. Antinociceptive Effects of Isosakuranetin in a Rat Model of Peripheral Neuropathy. Pharmacology. 2017;100(3-4):201-207.