Compatibility Attenuated Mechanism Of Nux Vomca Based On CYP3A4 Enzyme

Objective:The activity of CYP3 A4 enzyme was indicated by the concentration of midazolam which was measure through the HP LC method in rat’s liver microsomal.Explore the relationship between different combinations of NV and the activity of CYP3A4 enzyme.To clear the correlation among the activity,the protein content and the gene level of CYP3A4 enzyme by Western blot and RT-PCR technology.Meanwhile,determine the effect on the content and gene expression of nuclear receptor protein to explore the molecular mechanism of toxicity reduction of different combinations of N V,provide a theoretical basis for the clinical use of NV.Methods:Rats were randomLy divided into 8 groups:NA group,PRA group,PRA+NV group,R D group,RD+NV group,CCC group,CCC+NV group,RG group,RG+NV group and blank group.Evaluate the activity of CYP3 A4 enzyme by the change of the content of midazolam i n different groups.Western blot technology and RT-PCR technology were used to determine the protein content of CYP3A4 enzyme and the m RN A expression level in the groups that could induce the activity of CYP3A4 enzyme.And analyze the relationship of the active the enzyme,the protein and gene expression of the enzyme.Finally,we also detected the protein content and mRNA expression level of two nu clear receptors which are related to the CYP3 A4 enzyme by Western blot technology and RT-PCR technology.Explore the relationship between the changes of the protein content and the mRNA expression level of nuclear receptors and CYP3 A4 enzyme.Results:1.The results of the activity of CYP3A4 enzyme:compare with the blank group,the activity of CYP3A4 enzyme had no significant change in N A group,CCC group and CCC+NV group,while the activity can be increased in PR A group,PRA+NV group,RD group,RD+NV group,RG group and RG+NV group.2.The results of the protein content and mRN A expression level of CYP3A4 enzyme in the group that could induce the activity of CYP3A4 enzyme:(1)The results of the protein content of CYP3 A4 enzyme:compare with the blank group,the protein content of CYP3A4 enzyme had no significant change in NA group,while the protein content can be increased in PRA group,PRA+N V grou p,RD group,RD+NV group,RG group and RG+NV group.(2)The results of the mRNA expression level of CYP3A4 enzyme:compare with the blank group,the mRNA expression of CYP3A4 enzyme had no significant change in NA group,while the mRNA expression can be increased in PRA group,PRA+NV group,RD group,RD+NV group,RG group and RG+NV group.3.The results of the protein content and m RN A expression level of nuclear receptors in the group that could induce the activity of CYP3A4 enzyme:(1)The results of the protein content of nuclear receptors:compare with the blank group,PRA group,PRA+NV group,RD group and RD+NV group could induce the protein content of PXR,while there was no effect on the protein content of CAR.And RG group and RG+NV group had no change in the protein content of PXR and CAR.(2)The results of the mRNA expression of nuclear receptors:compare with the blank group,PR A group,RD group and PRA+NV group could induce the m RN A expression level of PXR,while there was no effect on the m RN A expression level of CAR.And RG group,RG+NV group and RD+NV group had no change in the m RN A expression level of PXR and CAR.Conclusions:1.In view of the metabolic enzymes,PRA RD CCC and after combine with N V respectively induces the activity of CYP3A4 enzyme and accelerates the metabolism of the toxic component in NV,thus achieve the purpose of reducing toxicity.2.According to the result of the activity of CYP3A4 enzyme,the protein content of CYP3 A4 enzyme and the m RN A expression level of CYP3A4 enzyme,the changes in the 3 parts had a certain correlation.The influence of the protein content and the gene expression may be one of the attenuated mechanisms.3.Compare the change of nuclear receptors and the protein content,the mRN A gene expression level of CYP3 A4 enzyme,the results showed that the effects on the activity of CYP3A4 and PXR were consistent in PRA group,RD group and PRA+NV group.RD+NV group could increase the protein content of PXR,but no effect on the m RN A expression level of PXR.It is deduced that RD and PR A can increase the protein content of CYP3A4 enzyme by up regulating of the mRNA expression of PXR protein,and then enhance the activity of CYP3 A4.