iTRAQ technology combined with 2D-LC-MS/MS to analyze effect of Coptidis Rhizoma on cytochrome P450 isoenzyme expression / 中国中药杂志

ABSTRACT

Coptidis Rhizoma is commonly used in clinical medicine. It has been widely used in traditional Chinese medicine (TCM), with the functions of clearing heat, drying dampness, purging intense heat relieve toxins. However, the herb contains complex ingredients, and CYP450 isoenzyme is the main metabolic enzyme of the drug, so it is of important clinical significance to study the effect of Coptidis Rhizoma on cytochrome P450 enzymes. In the experiment, liver tissues of rats were selected and liver microsomes were separated by differential centrifugation. Bicinchoninic acid (BCA) Protein quantitation was done by enzyme linked immunosorbent assay, and iTRAQ (isobaric tags for relative and absolute quantification) was combined with the 2D-LC-MS/MS analysis method for identification of CYP450 proteins. With Coptidis Rhizoma, 30 CYP450 isoenzymes were identified, with 7 proteins significantly increased and 8 decreased in response to Coptidis Rhizoma, while the rest 15 had no change. iTRAQ technology combined with 2D-LC-MS/MS method could be used to comprehensively study CYP450 enzyme, but it is necessary to further evaluate in vitro levels of Coptidis Rhizoma and avoid any potential clinical drug-drug interaction.