Study On The Effective On Cytochrome P450 And Hetapic Mechanism In Vinegar Processed Curcuma

Rhizoma curcumae is the dry roots of Curcuma phaeocaulis Val.、Curcuma kwangsiensis S.G.Lee et C.F.Liang、Curcuma wenyujin Y.H.Chen et C.Ling.This subject comes from the major projects of the university science research in Jiangsu Province:The research of concocted mechanism and Chinese medicine processed by vinegar on the role of liver.Early studies had showed that vinegar Rhizoma curcumae could promote curdione in rat liver tissue distribution,and the concentrations of curcumin in liver tissue was significantly higher than other major organizations after giving rats curcumin turmeric extract.Therefore,this research was mainly to compare the cell viability、the impact on CYP450 and biliary excretion of Rhizoma curcumae and vinegar Rhizoma curcumae.At the same time,used the new technology to discuss the effect of protecting liver with Rhizoma curcumae,and reveal the system theory of "processing with rice vinegar for leading the drug effect on liver".It will provide the basic knowledge for safe,effective and reasonable use in clinical.Curcuma Pieces was preparated and identificated in accordance with Chinese Pharmacopoeia.The results showed that the volatile oil content from seven batches was between 1.29%to 1.96%.The selected samples could be used for the experiments of the late.The cell viability of Rhizoma curcumae and vinegar Rhizoma curcumae was measured using the MTT assay.And coomassie blue method was used for the determination of the protein content of the cells.The results showed that there had significant differences in the cell viability with the concentration of 2 mg/ml、3 mg/ml,and the cell viability was higher with concentration of 2 mg/ml.The results suggested that vinegar Rhizoma curcumae could reduce cytotoxicity.The results of determination of the protein content of the cells showed that the protein content decreased as the concentration increased and the protein content was higher with concentration of 2 mg/ml.The hepatic coefficient,microsomal protein content and CYP450 content of mice was determined by UV.The results showed that Rhizoma curcumae could reduce liver coefficient and the role increased as the dose enhanced.Vinegar Rhizoma curcumae also could reduce liver coefficient and the role of it was stronger than Rhizoma curcumae.In addition to the low-dose group of Rhizoma curcumae,other dose groups of Rhizoma curcumae and vinegar Rhizoma curcumae could increase microsomal protein content,and the role of vinegar Rhizoma curcumae was stronger than Rhizoma curcumae.The high dose group of Rhizoma curcumae、the high and middle dose group of vinegar Rhizoma curcumae improved the CYP450 content significantly,and the role of vinegar Rhizoma curcumae was stronger than Rhizoma curcumae.The effects of Rhizoma curcuma and vinegar Rhizoma curcuma on mRNA expression of CYP1A2,CYP2E1 and CYP3A4 in rats liver was detected by QRT-PCR.The results of single dose showed that there was insignificant difference in the effects on mRNA expression of CYP1A2.CYP2E1 and CYP3A4 in rats liver between the blank group and the Rhizona curcunma group.Compared with the blank group,the expression of CYP1A2,CYP2E1 significantly decreased and the texpression of CYP3A4 increased significantly in the vinegar Rhizoma curcuma group.The results of multiple dose showed that there was insignificant difference in the effects on mRNA expression of CYP1A2 and CYP2E1 in rats liver between the blank group and the Rhizoma curcuma group.But the Rhizoma curcuma could up-regulated the expression of CYP3A4.The vinegar Rhizoma curcuma could significantly increase the expression of CYP3A4.It also decreased the expression of CYP1A2 and CYP2E1.It could be seen the role of the effects on mRNA expression of CYP CYP3A4 in the vinegar Rhizoma curcuma group was stronger than the Rhizoma curcuma group.The effects on cytochrome P450 isoforms activity of rats was measured by a three-drug Cocktail approach.The results of single dose showed that there was insignificant difference of the theophylline、dapsone、chlorzoxazone between the Rhizoma curcuma group and the blank group;Compared with the blank group,the half-life time、Tmax and the area under curve increased significantly and the clearance lower significantly of theophylline and chlorzoxazone in the vinegar Rhizoma curcuma group;There was insignificant difference in the half-life time of dapsone,the area under curve lower significantly and the clearance increased significantly in the vinegar Rhizoma curcuma group.It means that Rhizoma curczuma has no significant difference on the cytochrome P450 isoforms of CYP1A2、CYP3A4 and CYP2E1,it may be provided with the single dose:But the vinegar Rhizoma curcuma can inhibit CYP1A2 and CYP2E1 and has an induction on the CYP3A4.The results of multiple dose showed that compared with the blank group,the AUC0-24h and AUC0-x of theophylline in Rhizoma curcuma and vinegar Rhizoma curczuma groups increased,CL/F of theophylline decreased.Compared with the blank group,the AUC0-24h and AUC0-x of dapsone in Rhizoma curcuma and vinegar Rhizoma curcuma groups decreased.CL/F of dapsone increased.Compared with the blank group,the AUC0-x of chlorzoxazone in vinegar Rhizoma curcuma group increased,CL/F of chlorzoxazone decreased.It reflects that both Rhizoma curcuma and vinegar Rhizoma curcuma can inhibit CYP1A2 and has an induction on the CYP3A4 and the role of vinegar Rhizoma curcuma is stronger than Rhizoma czurcuma.Vinegar Rhizoma curcuma also can inhibit CYP2E1 enzyme activity and Rhizoma curcuma has no effect on CYP2E1.According to the existing research,the effects of Rhizoma curcuma and vinegar Rhizoma curcuma on the total biliary excretion after giving drug 12 h was measured by common bile duct intubation.The results showed that vinegar Rhizoma curcuma could promote biliary excretion.In summary,vinegar Rhizoma curcuma had more application than Rhizoma curcuma in clinical.This subject selected Rhizoma curcuma and vinegar Rhizoma curcuma as studied object and studied the cell viability.the role on CYP450 and the total biliary excretion of them.The results provide theoretical supporting for the vinegar processed of Rhizoma curcuma.It also provide the basic knowledge for safe,effective and reasonable use in clinical.