Xyloketal B, a marine compound, acts on a network of molecular proteins and regulates the activity and expression of rat cytochrome P450 3a: a bioinformatic and animal study

Su, Junhui; Chang, Cui; Xiang, Qi; Zhou, Zhi-Wei; Luo, Rong; Yang, Lun; He, Zhi-Xu; Yang, Hongtu; Li, Jianan; Bei, Yu; Xu, Jinmei; Zhang, Minjing; Zhang, Qihao; Su, Zhijian; Huang, Yadong; Pang, Jiyan; Zhou, Shu-Feng

The effect of XKB on rat hepatic Cyp3a activity was examined using the luminescent assay (P450-Glo) according to the manufacturer's instructions. Briefly, rat liver homogenates were prepared as described previously. 29 The reaction mixture (25 μL) was added to a 96-well microtiter plate containing microsomal protein (1 μg), Luciferin-IPA substrate (16 μM) , and 200 mM potassium phosphate buffer solution (pH 7.4), and mixed gently. The plate was preincubated at 37°C for 10 minutes. The reaction was initiated by adding 25 μL NADPH and the mixture was incubated at 37°C for 10 minutes. Next, 50 μL of reconstituted Luciferin detection reagent was added to all wells. The reaction mixture was mixed on a plate shaker for 10 seconds. The plate was then incubated at 37°C for 20 minutes. Cyp3a activity was examined by luminescence reading. The relative Cyp3a activity was expressed as fold change over the control.