Evaluation Of Impact Of Chinese Herbal Medicines On Liver Microsome Cytochrome P450 Enzymes With Multiple Reaction Monitoring Mass Spectrometry

Most of the drugs were metabolized through the drug metabolic enzymes CYP450 s in liver. Different drugs had different effects on metabolism enzymes CYP450 s, but the researches of traditional Chinese medicine on metabolism enzymes method were less efficient. The research on quantifying the drug metabolism enzyme CYP450 were as follows: 1) based on the western-blot method; 2) based on real-time quantitative polymerase chain reaction approach; 3) based on the cocktail probe substrates approach.These methods for CYP450 quanification were either non-specific nor low- accurate,Therefore, we need to establish a new specific and high-throughput method for CYP450 quanification, We successfully demonstrated an application of mass spectrometry with multiple reaction monitoring(MRM) to quantify eleven liver enzyme CYP450 isoforms peptides from the rat models which fed with traditional Chinese Herbal medicine such as decoction of salvia miltiorrhiza, Radix paeoniae alba, itum carmichaeli,Shenfu,Panax ginseng. With Qcon CAT heavy peptides as internal standard, we aimed to assess the effection on CYP450 that caused by the five herbs.Results showed the characterization of Qcon CAT peptides are same with rat liver microsomal peptides in mass spectrometry.The target protein abundance were quanified according to this strategy. the method acurracy and prescion were less than 3.85% and 4% and had a good linerarity(r2>0.9). The eleven quanified CYP450 isoforms were CYP1A1,CYP1A2,CYP2B1,CYP2B2,CYP2C6,CYP2C11,CYP2D1,CYP3A1,CYP3A2, CYP17A1 and CYP2E1,with their abundance were 15.12,14.43,5.63,2.37,6.05,18.27,16.24,4.78,48.2,69.32,66.66fmol/μg protein in normal rat liver microsome, The results revealed that CYP450 isoforms were regulated by Salvia miltiorrhiza, Radix paeoniae alba, Itum carmichaeli, Shenfu and Panax ginseng. Salvia miltiorrhiza had inhibitory effect on CYP1A1, CYP2B2 and induced effect on CYP1A2, CYP2B1. Panax ginseng inhibited the expression of the CYP3A2, CYP2C11 and CYP17A1. Radix paeoniae alba inhibited the expression of CYP1A1, CYP2C6, CYP2C11, CYP3A2 and induced the expression of CYP1A2 and CYP2D1. the expression of CYP2C6, CYP2C11, CYP3A2, CYP17A1 were inhibited by itum carmichaeli, CYP1A1 and CYP2D1 otherwise. And Shenfu had inhibitory effect on the expression of CYP1A1, CYP2C6, CYP2C11, CYP2E1,CYP3A2, and induced effect on the expression of CYP1A2 and CYP3A1. And we quantified the abundance of CYP3A4 in primary hepatocytes(0.53 fmol/μg), proveing that level of CYP3A4 was increased 1.6 times compared with control. Which we could provide clinical application for drug compatibility in practice and avoid adverse drug reactions.