Study On The Herb-drug Interactions Of Danhong Injection Based On CYP450 And UGT

Objective:To study the inhibition or induction of Danhong injection on drug metabolic enzyme CYP450 and UGT.Methods:（1）The concentrations of nine kinds of metabolites were respectively detected by LC-MS/MS after incubation in human liver microsome,and the half inhibitory concentration of DHI were calculated on CYP1A2,CYP2A6,CYP2B6,CYP2C8,CYP2C9,CYP2C19,CYP2D6,CYP2E1 and CYP3A4.（2）The concentrations of metabolites of CYP1A2,CYP2B and CYP3A were respectively detected by LC-MS/MS after incubation in human and rat primary hepatocytes and the relative induction percentage of DHI were calculated on enzyme subtypes.（3）CYP3A4 expression level was detected by RT-PCR after the human primary hepatocytes were induced by DHI and positive inducer respectively.（4）The concentration of 7-hydroxyl coumarin-D-glucuronic acid was detected by LC-MS/MS in human primary hepatocytes after incubation and the half inhibitory concentration of DHI was calculated on UGT.Results:（1）IC₅₀ value of Danhong injection on nine subtypes of CYP450 were0.793%,0.262%,0.665%,0.445%,1.181%,0.649%,0.676%,0.917%and 0.538%respectively.（2）The induction of CYP3A activity in rat and human primary hepatocytes by Danhong injection reached 20%and 30%of the positive inducer（rifampicin）.（3）The level of CYP3A4 mRNA was increased up to 4.1 and 6.7 folds by Danhong injection and rifampicin respectively.The induction of Danhong injection was up to 60%of the positive inducer.（4）The inhibition ratios of different concentrations（0.00%,0.01%,0.04%,0.12%,0.37%,1.11%,3.33%and 10.00%）Danhong injection on UGT were 0,-1.6%,3.2%,8.2%,5.2%,17.1%,38.3%and 71.4%respectively with concentration dependent inhibition.IC₅₀ value of Danhong injection on UGT was 4.7%.Conclusion:（1）The IC₅₀ of DHI for CYP2A6 was 0.262%,which was far less than the daily minimum concentration of DHI,so it has strong inhibitory effect.DHI has a moderate inhibitory effect on CYP1A2,CYP2B6,CYP2C8,CYP2C9,CYP2C19,CYP2D6,CYP2E1 and CYP3A4.（2）The induction effect of Danhong injection on CYP3A4 is up to 60%of the positive control drug,which has a certain induction effect.（3）Danhong injection has only a weak inhibitory effect on UGT with common doses in human hepatocytes and it is unlikely to cause the interaction of UGT related drugs.